Note: Descriptions are shown in the official language in which they were submitted.
WO 2023/288294
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COMPOSITIONS AND METHODS FOR IMPROVING
THE RAINFASTNESS OF PROTEINS ON PLANT SURFACES
RELATED APPLICATIONS
This application claims priority to US_ Provisional Application Nos.
63/222,612, filed July 16, 2021; 63/222,620,
filed July 16, 2021; and 63/342,064, filed May 14, 2022, the disclosure of
each of which is incorporated herein by reference
in its entirety.
REFERENCE TO A SEQUENCE LISTING
The application contains a Sequence Listing in computer readable form, which
is incorporated herein by reference
in its entirety.
FIELD OF THE INVENTION
The present disclosure relates to compositions and methods for improving the
rainfastness of proteins, including
enzymes, on the surfaces of plants and plant parts.
BACKGROUND
Myriad chemical compounds are currently approved for use as phytoprotective
agents. However, due to the
ecotoxicological effects of many of these agrochemicals, it is highly
desirable to develop effective and environmentally
friendly alternatives.
Protein-based actives, such as enzymes, cell-signaling proteins and
antibodies, are particularly desirable because
they are capable of exerting very specific and very effective phytoprotective
activities and because they are biodegradable
and will not accumulate in the environment even after many years of use.
Unfortunately, because proteins tend to be highly water-soluble, they are
susceptible to being washed off by rain
and/or irrigation events. In most instances, a rain or irrigation event that
occurs immediately after or soon after treatment
will remove all or substantially all the protein-based active from the surface
of the treated plant(s).
The present disclosure provides compositions and methods for improving the
rainfastness of a protein on a plant
surface (i.e., for increasing the likelihood the protein will remain on the
plant surface following a rain or irrigation event).
DETAILED DESCRIPTION
This description is not intended to be a detailed catalog of all the different
ways in which the inventive concepts
disclosed herein may be implemented or of all the features that may be added
thereto. For example, features illustrated
with respect to one embodiment may be incorporated into other embodiments and
features illustrated with respect to a
particular embodiment may be deleted from that embodiment. In addition,
numerous variations and additions to the various
embodiments suggested herein, which do not depart from the instant inventions,
will be apparent to those skilled in the art
in light of the instant disclosure. Hence, the following description is
intended to illustrate some embodiments of the instant
inventions and not to exhaustively specify all permutations, combinations and
variations thereof.
The terminology used herein is for the purpose of describing particular
embodiments only and is not intended to
be limiting of the instant inventions.
Unless otherwise defined, all terms (including technical and scientific terms)
used herein have the same meaning
as commonly understood by one of ordinary skill in the art to which the
inventions belong. It will be further understood
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that terms, such as those defined in commonly used dictionaries, should be
interpreted as having a meaning that is
consistent with their meaning in the context of the specification and relevant
art and should not be interpreted in an
idealized or overly fonnal sense unless expressly so defined herein. For the
sake of brevity and/or clarity, well-known
functions or constructions may not be described in detail.
As used herein, the singular forms "a," "an," and "the" are intended to
include the plural forms as well, unless the
context clearly indicates otherwise.
As used herein, "additive," when referring to effects of combinations within a
composition means that the effects
of the combinations are generally about the same as the sum of effects of the
individual components of the combination
alone. The combination of individual components producing this effect may be
called an additive combination.
As used herein, the term "agriculturally acceptable carrier" refers to a
substance or composition that can be used
to deliver an agriculturally beneficial agent to a plant, plant part or plant
growth medium (e.g., soil) without causing/having
an unduly adverse effect on plant growth and/or yield. As used herein, the
term "foliar-compatible carrier" refers to a
material that can be foliarly applied to a plant or plant part without
causing/having an unduly adverse effect on the plant,
plant part, plant growth, plant health, or the like. As used herein, the term
"seed-compatible carrier" refers to a material
that can be applied to a seed without causing/having an unduly adverse effect
on the seed, the plant that grows from the
seed, seed germination, or the like. As used herein, the term "soil-compatible
carrier" refers to a material that can be added
to a soil without causing/having an unduly adverse effect on plant growth,
soil structure, soil drainage, or the like.
As used herein, the term "agriculturally beneficial microorganism" refers to a
microorganism having at least one
agriculturally beneficial property (e.g., the ability to fix nitrogen, the
ability to solubilize phosphate and/or the ability to
produce an agriculturally beneficial agent, such as a plant signal molecule).
As used herein, the term "and/or" is intended to include any and all
combinations of one or more of the associated
listed items, as well as the lack of combinations when interpreted in the
alternative ("or"). Thus, the phrase "A, B and/or
C" is to be interpreted as "A, A and B, A and B and C, A and C, B, B and C, or
C."
As used herein, "antagonistic," when referring to effects of combinations
within a composition means that the
effects of the combinations are generally less than the sum of effects of the
individual components of the combination
alone. These compositions may be called antagonistic combinations.
As used herein, the term "aqueous" refers to a composition that contains more
than a trace amount of water (i.e.,
more than 0.5% water by weight, based upon the total weight of the
composition).
As used herein, the terms "associated with, in association with" and
"associated therewith," when used in
reference to a relationship between a composition of the present disclosure
and a plant or plant part, refer to at least a
juxtaposition or close proximity of the composition and the plant or plant
part. Such a juxtaposition or close proximity
may be achieved by contacting or applying the composition directly to the
plant or plant part and/or by applying the
composition to thc plant growth medium (e.g., soil) in which the plant or
plant part will be grown (or is currently being
grown). According to some embodiments, the composition is applied as a coating
to the outer surface of the plant or plant
part. According to some embodiments, the composition is applied to soil at,
near or surrounding the site in which the plant
or plant part will be grown (or is currently being grown).
As used herein, the term "biostimulant" refers to an agent or combination of
agents the application of which
enhances one or more metabolic and/or physiological processes of a plant or
plant part (e.g., carbohydrate biosynthesis,
ion uptake, nucleic acid uptake, nutrient delivery, photosynthesis and/or
respiration).
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As used herein, the term "binding module" refers to the region of an enzyme
that mediates binding to the enzyme
to a substrate.
As used herein, the term "catalytic domain" refers to the region of an enzyme
containing the catalytic machinery
of the enzyme.
As used herein, the term "cDNA" refers to a DNA molecule that can be prepared
by reverse transcription from a
mature, spliced, mRNA molecule obtained from a eukaryotic or prokaryotic cell.
cDNAs lack intron sequences that may
be present in the corresponding genomic DNA. The initial, primary RNA
transcript is a precursor to mRNA that is
processed through a series of steps, including splicing, before appearing as
mature spliced mRNA.
As used herein, the term "coding sequence" refers to a polynucleotide that
directly specifies the amino acid
sequence of a polypeptide. The boundaries of the coding sequence are generally
determined by an open reading frame,
which begins with a start codon, such as ATG, GTG, or TTG, and ends with a
stop codon, such as TAA, TAG, or TGA.
The coding sequence may be a genomic DNA, cDNA, synthetic DNA, or a
combination thereof.
As used herein, the terms "colony forming unit" and "cfu" refer to a microbial
cell/spore capable of propagating
on or in a suitable growth medium or substrate (e.g., a soil) when conditions
(e.g., temperature, moisture, nutrient
availability, pH, etc.) arc favorable for germination and/or microbial growth.
As used herein, the term "consists essentially of, when used in reference to
compositions and methods of the
present disclosure, means that the compositions/methods may contain additional
components/steps so long as the additional
components/steps do not materially alter the composition/method. The term
"materially alter," as applied to a
composition/method of the present disclosure, refers to an increase or
decrease in the effectiveness of the
composition/method of at least 20%. For example, a component added to a
composition of the present disclosure may be
deemed to "materially alter" the composition if it increases or decreases the
composition's ability to inhibit the growth of
a target phytopathogen by at least 20%.
As used herein, the term "control sequences" refers to nucleic acid sequences
involved in regulation of expression
of a polynucleotide in a specific organism or in vitro. Each control sequence
may be native (i.e., from the same gene) or
heterologous (i.e., from a different gene) to the polynucleotide encoding the
polypeptide, and native or heterologous to
each other. Such control sequences include, but are not limited to leader,
polyadenylation, prepropeptide, propeptide, signal
peptide, promoter, terminator, enhancer, and transcription or translation
initiator and terminator sequences. At a minimum,
the control sequences include a promoter, and transcriptional and
translational stop signals. The control sequences may be
provided with linkers for the purpose of introducing specific restriction
sites facilitating ligation of the control sequences
with the coding region of the polynucleotide encoding a polypeptide.
As used herein, the term "diazotroph" refers to an organism capable of
converting atmospheric nitrogen (N2) into
a form that may be utilized by a plant or plant part (e.g., ammonia (NH3),
ammonium (NH4+), etc.).
As uscd herein, thc term "dispersant" refers to an agent or combination of
agents the application of which reduces
the cohesiveness of like particles, the sulface tension of a liquid, the
intetfacial tension between two liquids and/or the
interfacial tension between or a liquid and a solid.
As used herein, the terms "effective amount," "effective concentration" and
"effective amount/concentration"
refer to an amount or concentration that is sufficient to cause a desired
effect (e.g.. inhibiting plant disease, enhancing
plant yield). The absolute value of the amount/concentration that is
sufficient to cause the desired effect may be affected
by factors such as the type and magnitude of effect desired, the type, size
and volume of material to which the
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composition will be applied, the type(s) of enzymes in the composition, the
amount(s) of enzyme(s) in the composition,
the stability of the enzyme(s) in the composition and the storage conditions
(e.g., temperature, relative humidity,
duration). Those skilled in the art will understand how to select an effective
amount/concentration using routine dose-
response experiments. In some examples, an effective amount of a substance
when used alone may be different than an
effective amount of the same substance when used as part of a combination.
As used herein, the terms "enhanced growth" and "enhanced plant growth" refer
to an improvement in one or
more characteristics of plant growth and/or development as compared to one or
more control plants (e.g., a plant
germinated from an untreated seed or an untreated plant). Exemplary plant
growth/development characteristics include,
but are not limited to, biomass, carbohydrate biosynthesis, chlorophyll
content, cold tolerance, drought tolerance, height,
leaf length, leaf mass, leaf number, leaf surface area, leaf volume, nutrient
uptake (e.g., calcium, magnesium, nitrogen,
phosphorous and/or potassium uptake), rate(s) of photosynthesis, root area,
root diameter, root length, root mass, root
nodulation (e.g., nodule mass, nodule number, nodule volume), root number,
root surface area, root volume, salt tolerance,
seed germination, seedling emergence, shoot diameter, shoot length, shoot
mass, shoot number, shoot surface area, shoot
volume, spread, stomatal conductance and survival rate.
As used herein, the terms "enhanced stability" and "enhanced enzyme stability"
refer to an improvement in one
or more characteristics of enzyme stability as compared to one or more
controls (e.g., a control composition that is identical
to a composition of the present disclosure except that it lacks one or more of
the components found in the composition of
the present disclosure). Exemplary enzyme stability characteristics include,
but are not limited to, maintenance of
enzymatic activity after being applied to a plant or plant part and/or stored
for a defined period of time and the ability to
cause a desired effect (e.g., reduced phy topathogenicity of a target pest)
after being applied to a plant or plant part and/or
stored for a defined period of time.
As used herein, the terms "enhanced yield" and 'enhanced plant yield" refer to
an improvement in one or more
characteristics of plant yield as compared to one or more control plants
(e.g., a control plant germinated from an untreated
seed). Exemplary plant yield characteristics include, but arc not limited to,
biomass; bushels per acre; grain weight per plot
(GWTPP); nutritional content; percentage of plants in a given area (e.g.,
plot) that fail to produce grain; yield at standard
moisture percentage (YSIvfP), such as grain yield at standard moisture
percentage (GYSIvfP); yield per plot (YPP), such
as grain weight per plot (GWTPP); and yield reduction (YRED).
As used herein, the term "expression" refers to any step involved in the
production of a polypeptide including,
but not limited to, transcription, post-transcriptional modification,
translation, post-translational modification, and
secretion. Expression can be measured¨for example, to detect increased
expression¨by techniques known in the art,
such as measuring levels of mRNA and/or translated polypeptide.
As used herein, the term "expression vector" refers to a linear or circular
DNA construct comprising a DNA
sequence encoding a polypeptide, which coding sequence is operably linked to a
suitable control sequence capable of
effecting expression of the DNA in a suitable host. Such control sequences may
include a promoter to effect transcription,
an optional operator sequence to control transcription, a sequence encoding
suitable ribosome binding sites on the mRNA,
enhancers and sequences which control termination of transcription and
translation.
As used herein, the term "extension" refers to an addition of one or more
amino acids to the amino and/or carboxyl
terminus of a polypeptide.
As used herein, the term "foliage" refers to those portions of a plant that
normally grow above the ground,
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including, but not limited to, leaves, stalks, stems, flowers, fruiting bodies
and fruits.
As used herein, the terms "foliar application" and "foliarly applied" refer to
the application of one or more active
ingredients to the foliage of a plant (e.g., to the leaves of the plant).
Application may be affected by any suitable means,
including, but not limited to, spraying/fogging the plant with a composition
comprising the active ingredient(s). In some
embodiments, the active ingredient(s) is/are applied to the leaves, stems
and/or stalk of the plant and not to the flowers,
fruiting bodies or fruits of the plant.
As used herein, the term "fragment" refers to a poly-peptide having one or
more amino acids absent from the
amino and/or carboxyl terminus of the mature polypeptide.
As used herein, the term "fusion polypeptide" refers to a polypeptide in which
one polypeptide is fused at the
N-terminus and/or the C-terminus of a polypeptide of the present disclosure. A
fusion polypeptide is produced by fusing
a polynucleotide encoding another polypeptide to a polynucleotide of the
present disclosure, or by fusing two or more
polynucleotides of the present disclosure together. Techniques for producing
fusion polypeptides are known in the art and
include ligating the coding sequences encoding the polypeptides so that they
are in frame and that expression of the fusion
polypeptide is under control of the same promoter(s) and terminator. Fusion
polypeptides may also be constructed using
intcin technology in which fusion polypeptides arc created post-
translationally (Cooper et at, 1993, EMBO J. 12: 2575-
2583; Dawson et al., 1994, Science 266: 776-779). A fusion poly-peptide can
further comprise a cleavage site between the
two polypeptides. Upon secretion of the fusion protein, the site is cleaved
releasing the two polypeptides. Examples of
cleavage sites include, but are not limited to, the sites disclosed in Martin
et al., 2003, J. Ind. Mtcrobtol. Btotechnol. 3:
568-576; Svetina et al., 2000, J. Biotechnol. 76: 245-251; Rasmussen-Wilson et
at., 1997, Appl. Environ. Microbiol. 63:
3488-3493; Ward et al., 1995, Biotechnology 13: 498-503; and Contreras el al.,
1991, Biotechnology 9: 378-381; Eaton el
al., 1986, Biochemistry 25: 505-512; Collins-Racie et al., 1995, Biotechnology
13: 982-987; Carter et at, 1989, Proteins:
Structure, Function, and Genetics 6: 240-248; and Stevens, 2003, Drug
Discovery World 4: 35-48.
As used herein, the term "heterologous," when used to describe the
relationship between a polynucleotide or
polypeptide and a host cell, refers to a polynucleotide or polypeptide that
does not naturally occur in the host cell.
As used herein, the term "heterologous," when used to describe the
relationship between a polynucleotide or
polypeptide and a control sequence (e.g., a promoter sequence), refers to a
polynucleotide or polypeptide is not naturally
associated with the control sequence (i.e., the control sequence is from a
gene other than the gene encoding the mature
polypeptide).
As used herein, the terms "host strain" and "host cell" refer to an organism
into which an expression vector, phage,
virus, or other DNA construct, including a polynucleotide encoding a
polypeptide of interest (e.g., an amylase) has been
introduced. Exemplary host strains are microorganism cells (e.g., bacteria,
filamentous fungi, and yeast) and plant cells
capable of expressing a protein of interest. The term "host cell" includes
protoplasts created from cells.
As uscd herein, the terms "inoculant composition" and "inoculum" refer to a
composition comprising microbial
cells and/or spores, said cells/spores being capable of
propagating/germinating on or in a suitable growth medium or
substrate (e.g., a soil) when conditions (e.g., temperature, moisture,
nutrient availability, pH, etc.) are favorable for
germination and/or microbial growth.
As used herein, the term "introduced," when used to describe the insertion of
a nucleic acid sequence into a cell,
encompasses "transfection", "transformation" or "transduction," as known in
the art.
As used herein, the term "isolated" refers to a polypeptide, nucleic acid,
cell, or other specified material or
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component that has been separated from at least one other material or
component, including but not limited to, other
proteins, nucleic acids, cells, etc. An isolated polypeptide, nucleic acid,
cell or other material is thus in a form that does
not occur in nature. An isolated polypeptide includes, but is not limited to,
a culture broth containing the secreted
polypeptide expressed in a host cell.
As used herein, the term "isomer" includes all stereoisomers of the compounds
and/or molecules to which it refers,
including enantiomers and diastereomers, as well as all conformers, roatmers
and tautomers, unless otherwise indicated.
Compounds and/or molecules disclosed herein include all enantiomers in either
substantially pure levorotatory or
dextrorotatory form, or in a racemic mixture, or in any ratio of enantiomers.
Where embodiments disclose a (D)-enantiomer,
that embodiment also includes the (L)-enantiomer; where embodiments disclose a
(L)-enantiomer, that embodiment also
includes the (D)-enantiomer. Where embodiments disclose a (+)-enantiomer, that
embodiment also includes the (-)-
enantiomer; where embodiments disclose a (-)-enantiomer, that embodiment also
includes the (+)-enantiomer. Where
embodiments disclose a (S)-enantiomer, that embodiment also includes the (R)-
enantiomer; where embodiments disclose
a (R)-enantiomer, that embodiment also includes the (S)-enantiomer.
Embodiments are intended to include any
diastereomers of the compounds and/or molecules referred to herein in
diastereomerically pure form and in the form of
mixtures in all ratios. Unless stereochemistry is explicitly indicated in a
chemical structure or chemical name, the chemical
structure or chemical name is intended to embrace all possible stereoisomers,
conformers, rotamers and tautomers of
compounds and/or molecules depicted.
As used herein, the term "mature polypeptide" refers to a polypeptide in its
mature form following N-terminal
and/or C-terminal processing (e.g., removal of signal peptide).
As used herein, the term "mature polypeptide coding sequence" refers to a
polynucleotide that encodes a mature
polypeptide.
As used herein, the term "modified microbial strain" refers to a microbial
strain that is modified from a strain
isolated from nature. Modified microbial strains may be produced by any
suitable method(s), including, but not limited to,
chemical or other form of induced mutation to a polynucleotide within any
genome within the strain; the insertion or
deletion of one or more nucleotides within any genome within the strain, or
combinations thereof; an inversion of at least
one segment of DNA within any genome within the strain; a rearrangement of any
genome within the strain; generalized
or specific transduction of homozygous or heterozygous polynucleotide segments
into any genome within the strain;
introduction of one or more phage into any genome of the strain;
transformation of any strain resulting in the introduction
into the strain of stably replicating autonomous extrachromosomal DNA; any
change to any genome or to the total DNA
composition within the strain isolated from nature as a result of conjugation
with any different microbial strain; and any
combination of the foregoing. The term modified microbial strains includes a
strain with (a) one of more heterologous
nucleotide sequences, (b) one or more non-naturally occurring copies of a
nucleotide sequence isolated from nature (i.e.,
additional copies of a gene that naturally occurs in thc microbial strain from
which the modified microbial strain was
derived), (c) a lack of one or more nucleotide sequences that would otherwise
be present in the natural reference strain by
for example deleting nucleotide sequence, and (d) added extrachromosomal DNA.
In some embodiments, modified
microbial strains comprise a combination of two or more nucleotide sequences
(e.g., two or more naturally occurring genes
that do not naturally occur in the same microbial strain) or comprise a
nucleotide sequence isolated from nature at a locus
that is different from the natural locus.
As used herein, the term "native" refers to a polynucleotide or polypeptide
naturally occurring in a host cell.
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As used herein, the term "naturally occurring" refers to anything (e.g.,
proteins, amino acids, or nucleic acid
sequences) that is found in nature. Conversely, the term "non-naturally
occurring" refers to anything that is not found in
nature (e.g., recombinant nucleic acids and protein sequences produced in a
laboratory, modification of a wild-type
sequence, formulations comprising one or more synthetic components,
formulations comprising an artificial combination
of otherwise naturally occurring components).
As used herein, the term "non-aqueous" refers to a composition that comprises
no more than a trace amount of
water (i.e., no more than 0.5% water by weight, based upon the total weight of
the composition).
As used herein, the term "nutrient" refers to a compound or element useful for
nourishing a plant (e.g., vitamins,
macrominerals, micronutrients, trace minerals, organic acids, etc. that are
necessary for plant growth and/or development).
As used herein, the term "polynucleotide" encompasses DNA, RNA,
heteroduplexes, and synthetic molecules
capable of encoding a polypeptide. Polynucleotides may be single-stranded or
double-stranded and may comprise chemical
modifications. The terms "nucleic acid" and "polynucleotide" are used
interchangeably. Because the genetic code is
degenerate, more than one codon may be used to encode a particular amino acid,
and the present compositions and methods
encompass nucleotide sequences that encode a particular amino acid sequence.
Unless otherwise indicated, nucleic acid
sequences arc presented in 5'-to-3' orientation.
As used herein, the term "nucleic acid construct" refers to a polynucleotide,
either single- or double-stranded,
which is isolated from a naturally occurring gene or is modified to contain
segments of nucleic acids in a manner that
would not otherwise exist in nature or which is synthetic, and which comprises
one or more control sequences operably
linked to the nucleic acid sequence.
As used herein, the term "operably linked" means that specified components are
in a relationship (including but
not limited to juxtaposition) permitting them to function in an intended
manner. For example, a regulatory sequence is
operably linked to a coding sequence such that expression of the coding
sequence is under control of the regulatory
sequence.
As used herein, the term "phosphate-solubilizing microorganism" refers to a
microorganism capable of converting
insoluble phosphate into a soluble fon of phosphate.
As used herein, the term "phytopathogenic pest" includes any organism or virus
that negatively affects a plant,
including, but not limited to, organisms and viruses that spread disease,
damage host plants and/or compete for soil
nutrients. The term "phytopathogenic pest" encompasses organisms and viruses
that are known to associate with plants
and to cause a detrimental effect on the plant's health and/or vigor.
Phytopathogenic pests include, but are not limited to,
arachnids (e.g., mites, ticks, spiders, etc.), bacteria, fungi, gastropods
(e.g., slugs, snails, etc.), invasive plants (e.g.,
weeds), insects (e.g., white flies, thrips, weevils, etc.), nematodes (e.g.,
root-knot nematode, soybean cyst nematode,
etc.), rodents and viruses (e.g., tobacco mosaic virus (TMV), tomato spotted
wilt virus (TSWV), cauliflower mosaic
virus (CaNIV), ctc.).
As used herein, the term "plant" includes all plant populations, including,
but not limited to, agricultural,
horticultural and silvicultural plants. The term "plant" encompasses plants
obtained by conventional plant breeding and
optimization methods (e.g., marker-assisted selection) and plants obtained by
genetic engineering, including cultivars
protectable and not protectable by plant breeders' rights.
As used herein, the term "plant cell" refers to a cell of an intact plant, a
cell taken from a plant, or a cell derived
from a cell taken from a plant. Thus, the term "plant cell" includes cells
within seeds, suspension cultures, embryos,
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meristematic regions, callus tissue, leaves, shoots, gametophytes,
sporophytes, pollen and microspores.
As used herein, the term "plant growth regulator refers to an agent or
combination of agents the application of
which accelerates or retards the growth/maturation rate of a plant through
direct physiological action on the plant or which
otherwise alters the behavior of a plant through direct physiological action
on the plant. "Plant growth regulator" shall not
be interpreted to include any agent or combination of agents excluded from the
definition of "plant regulator" that is set
forth section 2(v) of the Federal Insecticide, Fungicide, and Rodenticide Act
(7 U.S.C. 136(v)). Thus, "plant growth
regulator does not encompass microorganisms applied to a plant, plant part or
plant gmwth medium for the purpose of
enhancing the availability and/or uptake of nutrients, nutrients necessary to
normal plant growth, soil amendments applied
for the purpose of improving soil characteristics favorable for plant growth
or vitamin hormone products as defined by 40
C.F.R. 152.6(f).
As used herein, the term "plant part" refers to any part of a plant, including
cells and tissues derived from plants.
Thus, the term "plant part" may refer to any of plant components or organs
(e.g., leaves, stems, roots, etc.), plant tissues,
plant cells and seeds. Examples of plant parts, include, but are not limited
to, anthers, embryos, flowers, fruits, fruiting
bodies, leaves, ovules, pollen, rhizomes, roots, seeds, shoots, stems and
tubers, as well as scions, rootstocks, protoplasts,
calli and the like.
As used herein, the term "plant propagation material" refers to a plant part
from which a whole plant can be
generated. Examples of plant propagation materials include, but are not
limited to, cuttings (e.g., leaves, stems), rhizomes,
seeds, tubers and cells/tissues that can be cultured into a whole plant.
As used herein, the term "protein" is not meant to refer to a specific amino
acid chain length and encompasses
peptides, oligopeptides and polypeptides. It is to be understood that the term
"protein" also encompasses two or more
polypeptides combined to form an encoded product, as well as hybrid
polypeptides and fusion polypeptides.
As used herein, the term "purified" refers to a polynucleotide, protein or
cell that is substantially free from othcr
components as determined by analytical techniques well known in the art (e.g.,
a purified polynucleotide or protein may
form a discrete band in an clectrophoretic gel, chromatographic cluatc, and/or
a media subjected to density gradient
centrifugation). A purified polynucleotide or protein is at least about 50%
pure, usually at least about 60%, about 65%,
about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%,
about 93%, about 94%, about 95%,
about 96%, about 97%, about 98%, about 99%, about 99.5%, about 99.6%, about
99.7%, about 99.8% or more pure (e.g.,
percent by weight or on a molar basis). In a related sense, a composition is
enriched for a molecule when there is a
substantial increase in the concentration of the molecule after application of
a purification or enrichment technique. The
term "enriched" refers to a compound, polynucleotide, protein, cell, nucleic
acid, amino acid, or other specified material
or component that is present in a composition at a relative or absolute
concentration that is higher than a starting
composition.
ln onc aspcct, the term "purified" as uscd herein rcfcrs to the protein or
cell being essentially free from components
(especially insoluble components) from the production organism. In other
aspects. the term "purified" refers to the protein
being essentially free of insoluble components (especially insoluble
components) from the native organism from which it
is obtained. In one aspect, the protein is separated from some of the soluble
components of the organism and culture
medium from which it is recovered. The protein may be purified (i.e.,
separated) by one or more of the unit operations
filtration, precipitation, or chromatography.
Accordingly, the protein may be purified such that only minor amounts of other
proteins, in particular, other
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proteins, are present. The term "purified" as used herein may refer to removal
of other components, particularly other
proteins and most particularly other enzymes present in the cell of origin of
the protein. The protein may be "substantially
pure", i.e., free from other components from the organism in which it is
produced, e.g., a host organism for recombinantly
produced protein. In one aspect, the protein is at least 40% pure by weight of
the total protein material present in the
preparation. In one aspect, the protein is at least 50%, 60%, 70%, 80% or 90%
pure by weight of the total protein material
present in the preparation. As used herein. a "substantially pure protein" may
denote a protein preparation that contains at
most 10%, preferably at most 8%, more preferably at most 6%, more preferably
at most 5%, more preferably at most 4%,
more preferably at most 3%, even more preferably at most 2%, most preferably
at most 1%, and even most preferably at
most 0.5% by weight of other protein material with which the protein is
natively or recombinantly associated.
It is, therefore, preferred that the substantially pure protein is at least
92% pure, preferably at least 94% pure,
more preferably at least 95% pure, more preferably at least 96% pure, more
preferably at least 97% pure, more preferably
at least 98% pure, even more preferably at least 99% pure, most preferably at
least 99.5% pure by weight of the total
protein material present in the preparation. Proteins of the present
disclosure are preferably in a substantially pure form
(i.e., the preparations are essentially free of other protein material). This
can be accomplished, for example by preparing
the protein by well-known recombinant methods or by classical purification
methods.
As used herein, the tenn "recombinant is used in its conventional meaning to
refer to the manipulation, e.g.,
cutting and rejoining, of nucleic acid sequences to form constellations
different from those found in nature. The term
recombinant refers to a cell, nucleic acid, protein or vector that has been
modified from its native state. Thus, for example,
recombinant cells express genes that are not found within the native (non-
recombinant) form of the cell, or express native
genes at different levels or under different conditions than found in nature.
The term "recombinant" is synonymous with
"genetically modified" and "transgenic".
As used herein, the terms "recover" and "recovery" refer to the removal of a
protein from at least one fermentation
broth component selected from the list of a cell, a nucleic acid, or other
specified material, e.g., recovery of the protein
from the whole fermentation broth, or from the cell-frec fermentation broth,
by protein clystal harvest, by filtration, e.g.
depth filtration (by use of filter aids or packed filter medias, cloth
filtration in chamber filters, rotary-drum filtration, drum
filtration, rotary vacuum-drum filters, candle filters, horizontal leaf
filters or similar, using sheed or pad filtration in framed
or modular setups) or membrane filtration (using sheet filtration, module
filtration, candle filtration, microfiltration,
ultrafiltration in either cross flow, dynamic cross flow or dead end
operation), or by centrifugation (using decanter
centrifuges, disc stack centrifuges, hyrdo cyclones or similar), or by
precipitating the protein and using relevant solid-
liquid separation methods to harvest the protein from the broth media by use
of classification separation by particle sizes.
Recovery encompasses isolation and/or purification of the protein.
As used herein, the relatedness between two amino acid sequences or between
two nucleotide sequences is
described by thc parameter "sequence identity".
For pmposes of the present disclosure, the sequence identity between two amino
acid sequences is determined as
the output of "longest identity- using the Needleman-Wunsch algorithm
(Needleman and Wunsch, 1970, J. 'Viol. Biol. 48:
443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS:
The European Molecular Biology
Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably
version 6.6.0 or later. The parameters used
are a gap open penalty of 10, a gap extension penally of 0.5, and the
EBLOSUM62 (EMBOSS version of BLOSUM62)
substitution matrix. In order for the Needle program to report the longest
identity, the -nobrief option must be specified in
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the command line. The output of Needle labeled -longest identity" is
calculated as follows:
(Identical Residues x 100)/(Length of Alignment ¨ Total Number of Gaps in
Alignment)
For purposes of the present disclosure, the sequence identity between two
polynucleotide sequences is determined
as the output of "longest identity" using the Needleman-Wunsch algorithm
(Needleman and Wunsch, 1970, supra) as
implemented in the Needle program of the EMBOSS package (EMBOSS: The European
Molecular Biology Open
Software Suite, Rice et al., 2000, supra), preferably version 6.6.0 or later.
The parameters used are a gap open penalty of
10, a gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI
NUC4.4) substitution matrix. In order
for the Needle program to report the longest identity, the nobrief option must
be specified in the command line. The output
of Needle labeled "longest identity" is calculated as follows:
(Identical Deoxyribonucleotides x 100)/(Length of Alignment ¨ Total Number of
Gaps in Alignment)
As used herein, the term "signal peptide" refers to a sequence of amino acids
attached to the N-terminal portion
of a protein, which facilitates the secretion of the protein outside the cell.
The mature form of an extracellular protein lacks
the signal peptide, which is cleaved off during the secretion process.
As used herein, the terms "stabilizing compound" and "stabilizer" refer to an
agent or combination of agents the
application of which enhances the stability of an enzyme.
As used herein, the term "subsequence" refers to a polynucleotide having one
or more nucleotides absent from
the 5' and/or 3' end of a mature protein coding sequence; wherein the
subsequence encodes a fragment having enzymatic
activity.
As used herein, the term "variant- refers to a protein comprising a man-made
mutation, i.e., a substitution,
insertion (including extension), and/or deletion (e.g., truncation), at one or
more positions. A substitution means
replacement of the amino acid occupying a position with a different amino
acid; a deletion means removal of the amino
acid occupying a position; and an insertion means adding 1-5 amino acids
(e.g., 1-3 amino acids, in particular, 1 amino
acid) adjacent to and immediately following the amino acid occupying a
position.
As used herein, the term "wild-type" in reference to an amino acid sequence or
nucleic acid sequence means that
the amino acid sequence or nucleic acid sequence is a native or naturally
occurring sequence.
While certain aspects of the present disclosure will hereinafter be described
with reference to embodiments
thereof, it will be understood by those of ordinary skill in the art that
various changes in form and details may be made
therein without departing from the spirit and scope of the present disclosure
as defined by the claims.
All publications, patent applications, patents and other references mentioned
herein are incorporated by reference
in their entirety, except insofar as they contradict any disclosure expressly
set forth herein.
The present disclosure provides proteins useful for preventing, treating,
suppressing, eliminating and/or
reducing the severity of infestations/infections of/by various pests,
including, but not limited to, horticultural pests, such
as acarids, bacteria, fungi, gastropods, insects, nematodes, oomycctcs,
protozoa and weeds, as well as formulations
comprising such proteins, polynucleotides encoding such proteins, organisms
expressing such proteins, and methods of
using such proteins, formulations, polynucleotides and organisms in
agriculture and other fields of endeavor.
The present disclosure encompasses proteins having one or more catalytic
activities, including, but not limited
to, proteins capable of exhibiting one or more catalytic activities belonging
to Enzyme Commission classification
number 1 (EC 1), Enzyme Commission classification number 3 (EC 3) and/or
Enzyme Commission classification
number 4 (EC 4).
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In some embodiments, proteins of the present disclosure exhibit one or more
catalytic activities belonging to
EC 1. For example, in some embodiments, proteins of the present disclosure
exhibit glucose oxidase, cellobiose
dehydrogenase, laccase, catalase and/or peroxidase activity useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of infestations/infections of/by pests, such as
acarids, bacteria, fungi, gastropods, insects,
nematodes, oomycetes and protozoa.
In some embodiments, proteins of the present disclosure exhibit oxidoreductase
activity belonging to EC 1 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 1-12.
In some embodiments, proteins of the present disclosure exhibit one or more
oxidoreductase activities
belonging to EC 1.1, such as oxidase activities belonging to EC 1.1.3 (e.g.,
glucose oxidase activity belonging to EC
1.1.3.4 and/or cellobiose dehydrogenase activity belonging to EC 1.1.3.25).
in some embodiments, proteins of the present disclosure exhibit oxidoreductase
activity belonging to EC 1.1
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70. 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 1-8.
In some embodiments, proteins of the present disclosure exhibit oxidase
activity belonging to EC 1.1.3 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71. 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 1-8.
In some embodiments, proteins of the present disclosure exhibit glucose
oxidase activity belonging to EC
1.1.3.4 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 1-
5.
In some embodiments, proteins of the present disclosure exhibit cellobiose
dehydrogenase activity belonging to
EC 1.1.3.25 (now included in EC 1.1.99.18) and, optionally comprise, consist
essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75. 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90. 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 6-8.
In some embodiments, proteins of the present disclosure exhibit one or more
oxidoreductase activities
belonging to EC 1.10, such as activities belonging to EC 1.10.3 (e.g., laccase
activity belonging to EC 1.10.3.2).
In somc embodiments, protcins of the prcscnt disclosure exhibit oxidasc
activity belonging to EC 1.10 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to the amino acid sequences set forth herein as
SEQ ID NO: 9.
In some embodiments, proteins of the present disclosure exhibit oxidase
activity belonging to EC 1.10.3 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
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97, 98, 99 or 100 % identical to the amino acid sequences set forth herein as
SEQ ID NO: 9.
In some embodiments, proteins of the present disclosure exhibit laccase
activity belonging to EC 1.10.3.2 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to the amino acid sequences set forth herein as
SEQ ID NO: 9.
In some embodiments, proteins of the present disclosure exhibit one or more
oxidoreductase activities
belonging to EC 1.11, such as peroxidase activities belonging to EC 1.11.1
(e.g., catalase activity belonging to EC
1.11.1.6 and/or peroxidase activity belonging to 1.11.1.7).
In some embodiments, enzymes of the present disclosure exhibit peroxidase
activity belonging to EC 1.11 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 10-12.
in some embodiments, proteins of the present disclosure exhibit peroxidase
activity belonging to EC 1.11.1 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 10-12.
In some embodiments, proteins of the present disclosure exhibit catalase
activity belonging to EC 1.11.1.6 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 10-11.
In some embodiments, proteins of the present disclosure exhibit peroxidase
activity belonging to EC 1.11.1.7
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequences set forth herein
as SEQ ID NO: 12.
In some embodiments, proteins of the present disclosure exhibit one or more
ovgenase activities belonging to
EC 1.14, such as ovgenase activities belonging to 1.14.99 (e.g., lytic
cellulose monooxygenase activity belonging to EC
1.14.99.56).
In some embodiments, proteins of the present disclosure exhibit ovgenase
activity belonging to EC 1.14 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 98-99.
In some embodiments, proteins of the present disclosure exhibit ovgenase
activity belonging to EC 1.14.99
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 98-99.
In some embodiments, proteins of the present disclosure exhibit lytic
cellulose monooxygenase activity
belonging to EC 1.14.99.56 and, optionally, comprise, consist essentially of,
or consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
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herein as SEQ ID NOs: 98-99.
In some embodiments, proteins of the present disclosure exhibit one or more
catalytic activities belonging to EC
3. For example, in some embodiments, proteins of the present disclosure
exhibit lipase, triacylglycerol lipase,
pectinesterase, phospholipase, lysophospholipase, amylase, glucosidase,
galactosidase, cellulose, glucanase, xylanase,
ceramidase, dextranase, chitinase, chitosanase, galacturonase, fucosidase,
lysozymes, vlosidase, lucosidass, pullulanase,
mannosidase, amidase, asparaginase, aminidase, maltohydrolases,
cellobiosidase, pectinase, aminopeptidase, serine
peptidase and/or metallopeptidase activity useful for preventing, treating,
suppressing, eliminating and/or reducing the
severity of infestations/infections of/by pests, such as acarids, bacteria,
fungi, gastropods, insects, nematodes, oomycetes
and protozoa.
In some embodiments, proteins of the present disclosure exhibit esterase
activity belonging to EC 3 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ TD NOs: 13-48, 100-
148 and 150.
In some embodiments, proteins of the present disclosure exhibit one or more
esterase activities belonging to EC
3.1, such as lipase activities belonging to EC 3.1.1 (e.g., triacylglycerol
lipase activity belong to EC 3.1.1.3,
lysophospholipase activity belonging to 3.1.1.5, pectinesterase activity
belonging to 3.1.1.11, and/or phospholipase
activity belonging to 3.1.1.32) and hydrolase activities belonging to EC 3.1.4
(e.g., phospholipase C activity belonging to
3.1.4.3 and/or phoshoinositide phospholipase C activity belonging to
3.1.4.11).
In some embodiments, proteins of the present disclosure exhibit esterase
activity belonging to EC 3.1 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NO: 13 and 100-
108.
In some embodiments, proteins of the present disclosure exhibit esterase
activity belonging to EC 3.1.1 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 `)/0 identical to one or more of the amino acid sequences
set forth herein as SEQ ID NO: 13 and 100-
106.
In some embodiments, proteins of the present disclosure exhibit
triacylglycerol lipase activity belonging to EC
3.1.1.3 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NO:
100-103.
In some embodiments, proteins of the present disclosure exhibit
lysophospholipase activity belonging to EC
3.1.1.5 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NO: 13
and 104.
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In some embodiments, proteins of the present disclosure exhibit pectinesterase
activity belonging to EC 3.1.1.11
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NO: 100-103.
In some embodiments, proteins of the present disclosure exhibit phospholipase
A1 activity belonging to EC
3.1.1.32 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NO:
100-103.
In some embodiments, proteins of the present disclosure exhibit one or more
glycosylase activities belonging to
EC 3.2, such as glycosidase activities belonging to EC 3.2.1 (e.g., alpha-
amylase activity belong to EC 3.2.1.1, beta-
amylase activity belong to EC 3.2.1.2, glucan 1,4-alpha-glucosidase activity
belong to 3.2.1.3, cellulase activity belong
to 3.2.1.4, endo-1,3(4)-beta-glucanase activity belong to 3.2.1.6, inulinase
activity belong to 3.2.1.7, endo-1,4-beta-
xylanase activity belong to 3.2.1.8, oligo-1,6-glucosidase activity belong to
3.2.1.10, dextranase activity belong to
3.2.1.11, chitinasc activity belong to 3.2.1.14, endo-polygalacturonase
(pectinase) activity belong to 3.2.1.15, lysozymc
activity belong to 3.2.1.17, alpha-glucosidase activity belong to 3.2.1.20,
beta-glucosidase activity belong to 3.2.1.21,
alpha-galactosidase activity belong to 3.2.1.22, beta-galactosidase activity
belong to 3.2.1.23, alpha-mannosidase activity
belong to 3.2.1.24, beta-mannosidase activity belong to 3.2.1.25, beta-
furctofuranosidase activity belong to 3.2.1.26,
alpha,alpha-trehalase activity belong to 3.2.1.28, endo-1,3-beta-xylanase
activity belong to 3.2.1.32, xylan 1,4-beta-
xylosidase activity belong 1o3.2.1.37. glucan endo-1,3-beta-D-glucosidase
activity belong to 3.2.1.39, pullulanase
activity belong to 3.2.1.41, alpha-L-arabinofuranosidase activity belong to
3.2.1.55, glucan 1,3-beta-glucosidase activity
belong to 3.2.1.58, glucan endo-1,3-alpha-glucosidase activity belong to
3.2.1.59, licheninase activity belong to 3.2.1.73,
glucan endo-1,6-beta-glucosidase activity belong to 3.2.1.75, mannan endo-1,4-
13-mannosidase activity belonging to
3.2.1.78, 1,4-beta-cellobiosidase activity belong to 3.2.1.91, endo-alpha-N-
acetylgalactosaminidase activity belong to
3.2.1.97, mannan endo-1,6-alpha-mannosidase activity belong to 3.2.1.101,
endogalactosaminidase activity belong to
3.2.1.109, 1,3-alpha-L-fucosidase activity belong to 3.2.1.111, 2-
deoxyglucosidase activity belong to 3.2.1.112,
chitosanase activity belong to 3.2.1.132, glucan 1,4-alpha-maltohydrolase
activity belong to 3.2.1.133, and/or 1,4-beta-
cellobiosidase activity belonging to 3.2.1.176).
In some embodiments, proteins of the present disclosure exhibit glycosylase
activity belonging to EC 3.2 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 14-41 and
109-143.
In some embodiments, proteins of the present disclosure exhibit glycosidase
activity belonging to EC 3.2.1 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 14-41 and
109-143.
In some embodiments, proteins of the present disclosure exhibit alpha-amylase
activity belonging to EC 3.2.1.1
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and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 14-16 and
109-113.
In some embodiments, proteins of the present disclosure exhibit glucan 1,4-
alpha-glucosidase activity belonging
to EC 3.2.1.3 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 17-18.
In some embodiments, proteins of the present disclosure exhibit cellulase
activity belonging to EC 3.2.1.4 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ TD NO: 19 and 114-
116.
In some embodiments, proteins of the present disclosure exhibit endo-1,3(4)-
beta-glucanase activity belonging
to EC 3.2.1.6 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NO: 20 and 150.
In some embodiments, proteins of the present disclosure exhibit inulinase
activity belonging to EC 3.2.1.7 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to the amino acid sequence set forth herein as
SEQ ID NO: 117.
In some embodiments, proteins of the present disclosure exhibit endo-1,4-beta-
xylanase activity belonging to
EC 3.2.1.8 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs:
21-24 and 118-120.
In some embodiments, proteins of the present disclosure exhibit dextranase
activity belonging to EC 3.2.1.11
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth herein
as SEQ ID NO: 25.
In some embodiments, proteins of the present disclosure exhibit chitinasc
activity belonging to EC 3.2.1.14 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 121-124 and
141.
In some embodiments, proteins of the present disclosure exhibit endo-
polygalacturonase (pectinase) activity
belonging to EC 3.2.1.15 and, optionally, comprise, consist essentially of, or
consist of an amino acid sequence that is
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about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70. 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NO: 125.
In some embodiments, proteins of the present disclosure exhibit lysozyme
activity belonging to EC 3.2.1.17
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70. 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth herein
as SEQ ID NO: 26.
In some embodiments, proteins of the present disclosure exhibit beta-
glucosidase activity belonging to EC
3.2.1.21 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % identical to the amino acid sequence set
forth herein as SEQ ID NO: 126.
In some embodiments, proteins of the present disclosure exhibit xylan 1,4-beta-
xylosidase activity belonging to
EC 3.2.1.37 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98. 99 or 100 % identical to the amino acid sequence
set forth herein as SEQ ID NO: 127.
In some embodiments, proteins of the present disclosure exhibit glucan endo-
1,3-beta-D-glucosidase activity
belonging to EC 3.2.1.39 and, optionally, comprise, consist essentially of, or
consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 27-28 and 128.
In some embodiments, proteins of the present disclosure exhibit pullulanase
activity belonging to EC 3.2.1.41
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth herein
as SEQ ID NO: 29.
In some embodiments, proteins of the present disclosure exhibit alpha-L-
arabinofuranosidase activity belonging
to EC 3.2.1.55 and, optionally, comprise, consist essentially of, or consist
of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98. 99 or 100 % identical to the amino acid sequence
set forth herein as SEQ ID NO: 129.
In some embodiments, proteins of the present disclosure exhibit glucan endo-
1,3-alpha-glucosidase activity
belonging to EC 3.2.1.59 and, optionally, comprise, consist essentially of, or
consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 30 and 130-131.
In some embodiments, proteins of the present disclosure exhibit licheninase
activity belonging to EC 3.2.1.73
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NO: 132.
In some embodiments, proteins of the present disclosure exhibit glucan endo-
1,6-beta-glucosidase activity
belonging to EC 3.2.1.75 and, optionally, comprise, consist essentially of, or
consist of an amino acid sequence that is
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about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70. 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 31-32 and 133.
In some embodiments, proteins of the present disclosure exhibit mannan endo-
1,4-beta-mannosidase activity
belonging to EC 3.2.1.78 and, optionally, comprise, consist essentially of, or
consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 33 and 134.
In some embodiments, proteins of the present disclosure exhibit 1,4-beta-
cellobiosidase activity belonging to
EC 3.2.1.91 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to the amino acid sequences
set forth herein as SEQ ID NO: 135.
in some embodiments, proteins of the present disclosure exhibit mannan endo-
1,6-alpha-mannosidase activity
belonging to EC 3.2.1.101 and, optionally, comprise, consist essentially of,
or consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70. 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86. 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 34 and 136.
In some embodiments, proteins of the present disclosure exhibit
endogalactosaminidase activity belonging to
EC 3.2.1.109 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 35-40 and 137-138.
In some embodiments, proteins of the present disclosure exhibit chitosanase
activity belonging to EC 3.2.1.132
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 139-140.
In some embodiments, proteins of the present disclosure exhibit glucan 1,4-
alpha-maltohydrolase activity
belonging to EC 3.2.1.133 and, optionally, comprise, consist essentially of,
or consist of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to the amino
acid sequences set forth herein as SEQ ID
NO: 41.
In some embodiments, proteins of the present disclosure exhibit 1,4-beta-
cellobiosidase activity belonging to
EC 3.2.1.176 and, optionally, comprise, consist essentially of, or consist of
an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 142-143.
In some embodiments, proteins of the present disclosure exhibit one or more
peptidase activities belonging to
EC 3.4, such as aminopeptidase activities belonging to EC 3.4.11 (e.g., leucyl
aminopeptidase activity belong to EC
3.4.11.1), serine endopeptidase activities belonging to EC 3.4.21 (e.g.,
glutamylendopeptidase activity belonging to EC
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3.4.21.19 and/or subtilisin activity belonging to 3.4.21.62) and
metalloendopeptidase activity belonging to EC 3.4.24
(e.g., bacillolysin activity belonging to EC 3.4.24.28).
In some embodiments, proteins of the present disclosure exhibit peptidase
activity belonging to EC 3.4 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 42-48.
In some embodiments, proteins of the present disclosure exhibit serine
endoprotease activity belonging to EC
3.4.21 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs:
42-48 and 144-145.
In some embodiments, proteins of the present disclosure exhibit glutainyl
endoprotease activity belonging to EC
3.4.21.19 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % identical to the amino acid sequence set
forth herein as SEQ ID NO: 43.
In some embodiments, proteins of the present disclosure exhibit subtilisin
activity belonging to EC 3.4.21.62
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 A) identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 44-47 and
145.
In some embodiments, proteins of the present disclosure exhibit
metalloendopeptidase activity belonging to EC
3.4.24 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to the amino acid sequences set
forth herein as SEQ ID NO: 48.
In some embodiments, proteins of the present disclosure exhibit bacillolysin
activity belonging to EC 3.4.24.28
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequences set forth herein
as SEQ ID NO: 48.
In some embodiments, proteins of the present disclosure exhibit one or more
hydrolase activities belonging to
EC 3.5, such as amidohydrolase and amidase activities belonging to EC 3.5.1
(e.g., asparaginase activity belong to EC
3.5.1.1, glutaminase activities belonging to 3.5.1.2, amidase activities
belonging to 3.5.1.4, urease activities belonging to
3.5.1.5, biotinidase activities belonging to 3.5.1.12, nicotinamidase
activities belonging to 3.5.1.19, D-glutaminase
activities belonging to 3.5.1.35, glutamin-(asparagin-)ase activities
belonging to 3.5.1.38, chitin dcacetylase activities
belonging to 3.5.1.41, peptidyl-glutaminase activities belonging to 3.5.1.43,
protein-glutamine glutaminase activities
belonging to 3.5.1.44, pentanamidase activities belonging to 3.5.1.50).
In some embodiments, proteins of the present disclosure exhibit hydrolase
activity belonging to EC 3.5 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 146-148.
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In some embodiments, proteins of the present disclosure exhibit deacetylase
activity belonging to EC 3.5.1 and.
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to one or more of the amino acid sequences set
forth herein as SEQ ID NOs: 42-48 and
146-148.
In some embodiments, proteins of the present disclosure exhibit asparaginase
activity belonging to EC 3.5.1.1
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth herein
as SEQ ID NO: 148.
In some embodiments, proteins of the present disclosure exhibit one or more
lyase activities belonging to EC 4.
For example, in some embodiments, proteins of the present disclosure exhibit
lyase activity useful for preventing,
treating, suppressing, eliminating and/or reducing the severity of
infestations/infections of/by pests, such as acarids,
bacteria, fungi, gastropods, insects, nematodes, oomycetes and protozoa.
In some embodiments, proteins of the present disclosure exhibit one or more
lyase activities belonging to EC
4.2, such as polysaccharide lyase activities belonging to EC 4.2.2 (e.g.,
pectate lyase activity belong to EC 4.2.2.2, pectin
lyase activity belonging to EC 4.2.2.10, glucan lyase activity belonging to EC
4.2.2.13, gellan lyase activity belonging to
EC 4.2.2.25, oligo-alginase lyase activity belonging to EC 4.2.2.26, pectin
monosaccharide-lyase activity belonging to
EC 4.2.2.27).
In some embodiments, proteins of the present disclosure exhibit lyase activity
belonging to EC 4.2 and,
optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98, 99 or 100 % identical to the amino acid sequence set forth herein as
SEQ ID NO: 149.
In some embodiments, proteins of the present disclosure exhibit polysaccharide
lyase activity belonging to EC
4.2.2 and, optionally, comprise, consist essentially of, or consist of an
amino acid sequence that is about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth
herein as SEQ ID NO: 149.
In some embodiments, proteins of the present disclosure exhibit pectate lyase
activity belonging to EC 4.2.2.2
and, optionally, comprise, consist essentially of, or consist of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to the amino acid sequence set forth herein
as SEQ ID NO: 149.
It is to be understood that the present disclosure extends to proteins capable
of exhibiting two, three, four, five
or more distinct catalytic activities (e.g., a fusion protein comprising a
first polypeptide having a first catalytic activity
and a sccond polypcptidc having a sccond catalytic activity).
It is to be further understood that proteins of the present disclosure needn't
be toxic to be effective. As will be
explained in further detail below, proteins of the present disclosure may
exert their effects through various non-lethal
means, such as reducing the attraction of a pest to a treated surface by
degrading a food source, for example. Moreover,
in many instances, otherwise toxic proteins of the present disclosure may be
used in non-lethal doses to enhance the
efficacy of and/or expand the target pest range of various chemical pesticides
and biological pesticides.
Table 1 provides a non-exhaustive list of proteins useful for preventing,
treating, suppressing, eliminating
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and/or reducing the severity of infestations/infections of/by one or more
pests, such as acarids, bacteria, fungi, insects,
nematodes, oomycetes and protozoa.
Table 1. Exemplary proteins of the present disclosure
Polypeptide
Polynucleotide
NZ # Enzyme Class EC #
SEQ ID NO: SEQ ID NO:
1 glucose oxidase 1.1.3.4 1 49
2 glucose oxidase 1.1.3.4 2 50
3 glucose oxidase 1.1.3.4 3 51
4 glucose oxidase 1.1.3.4 4 52
glucose oxidase 1.1.3.4 5 53
6 cellobiose oxidase 1.1.3.25 6 54
7 cellobiose oxidase 1.1.3.25 7 55
8 cellobiose oxidase 1.1.3.25 8 56
9 laccase 1.10.3.2 9 57
catalase 1.11.1.6 10 58
11 catalase 1.11.1.6 11 59
11 catalase 1.11.1.6 11 60
12 peroxidase 1.11.1.7 12 61
99 lytic cellulose monooxygenase 1.14.99.56 99 152
100 triacylglycerol lipase 3.1.1.3 100
153
101 triacylglycerol lipase 3.1.1.3 101
154
102 triacylglycerol lipase 3.1.1.3 102
155
103 triacylglycerol lipase 3.1.1.3 103
156
13 lysophospholipase 3.1.1.5 13 62
105 pectinestemse 3.1.1.11 105
158
14 alpha-a myla se 3.2.1.1 14 63
alpha-amylase 3.2.1.1 15 64
16 alpha-amylase 3.2.1.1 16 65
109 alpha-amylase 3.2.1.1 109
162
110 alpha-amylase 3.2.1.1 110
163
111 alpha-amylase 3.2.1.1 111
164
17 glucan 1,4-alpha-glucosidase 3.2.1.3 17 66
18 glucan 1,4-alpha-glucosidase 3.2.1.3 18 67
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19 cellulase 3.2.1.4 19
68
114 cellulase 3.2.1.4 114
167
116 cellulase 3.2.1.4 116
169
20 endo-1,3(4)-beta-glucanase 3.2.1.6 20
69
117 inulinasc 3.2.1.7 117
170
21 endo-1,4-beta-xylanase 3.2.1.8 21
70
22 endo-1,4-beta-vlanase 3.2.1.8 22
71
23 cndo-1,4-bcta-xylanasc 3.2.1.8 23
72
24 endo-1,4-beta-vlanase 3.2.1.8 24
73
119 endo-1,4-beta-xylanase 3.2.1.8 119
172
25 dcxtranasc 3.2.1.11 25
74
125 endo-polygalacturonase (pectinase) 3.2.1.15 125
178
26 lysozyme 3.2.1.17 26
75
126 beta-glucosidase 3.2.1.21 126
179
27 glucan endo-1,3-beta-glucosidase 3.2.1.39 27
76
28 glucan endo-1,3-beta-glucosidase 3.2.1.39 28
77
29 pullulanase 3.2.1.41 29
78
129 alpha-L-arabinofuranosidase 3.2.1.55 129
182
30 glucan endo-1,3-alpha-glucosidase 3.2.1.59 30
79
132 licheninase 3.2.1.73 132
185
31 glucan endo-1,6-beta-glucosidase 3.2.1.75 31
80
32 glucan endo-1,6-beta-glucosidase 3.2.1.75 32
81
33 mannan endo-1,4-beta-mannosidase 3.2.1.78 33
82
34 mannan endo-1,6-alpha-mannosidase 3.2.1.101 34
83
35 endogalactosaminidase 3.2.1.109 35
84
36 endogalactosaminidase 3.2.1.109 36
85
37 endogalactosaminidase 3.2.1.109 37
86
38 endogalactosaminidase 3.2.1.109 38
87
39 endogalactosaminidase 3.2.1.109 39
88
40 endogalactosaminidase 3.2.1.109 40
89
41 glucan 1,4-alpha-maltohydrolase 3.2.1.133 41
90
42 serine endopeptidase 3.4.21 42
91
43 glutamyl endopeptidase 3.4.21.19 43
92
44 subtilisin 3.4.21.62 44
93
45 subtilisin 3.4.21.62 45
94
46 subtilisin 3.4.21.62 46
95
47 subtilisin 3.4.21.62 47
96
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48 bacillolycin 3.4.24.28 48
97
149 pectate lyase 4.2.2.2 149
202
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100% sequence identity to SEQ TD NO:
1;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 'A sequence identity to a mature
polypeptide of SEQ ID NO: 1;
c) a polypeptide encoded by a polynucleolide having about/al
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 49 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 1 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
1 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through c) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucose oxidase activity.
in some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70. 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100% sequence identity to SEQ ID NO:
2;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 'A sequence identity to a mature
polypeptide of SEQ ID NO: 2;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 50 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 2 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
2 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
3;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 3;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 51 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 3 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
3 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
4;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 4;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 52 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 4 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
4 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
5;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 5;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 53 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 5 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 5 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
6;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 6;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 54 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 6 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
6 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellobiose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
7;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 7;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 55 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 7 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 7 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellobiose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
8;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 8;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 56 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 8 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
8 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellobiose oxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
9;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 9;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 57 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 9 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 9 by
substitution, deletion or insertion of
one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has laccase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
10;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 10:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 58 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 10 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
10 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has catalase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
11;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 11:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 59 or the cDNA sequence thereof;
d) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 60 or the cDNA sequence thereof;
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e) a polypeptide derived from SEQ ID NO: 11 by substitution,
deletion or insertion of one or more amino
acids;
0 a polypeptide derived from a mature polypeptide of SEQ ID NO:
11 by substitution, deletion or insertion of
one or more amino acids;
g) a polypeptide derived from the polypeptide of any one of a) through f)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
h) a fragment of the polypeptide of any one of a) through g)
wherein the polypeptide has catalase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
12;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 12:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 61 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 12 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
12 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has peroxidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
13;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 13:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 62 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 13 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
13 by substitution, deletion or insertion of
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one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through 1)
wherein the polypeptide has lysophospholipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
14;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 14;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 63 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 14 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
14 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
15;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 15;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 64 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 15 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
15 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
16;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 16;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 65 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 16 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
16 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
17;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 17;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 66 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 17 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
17 by substitution, deletion or insertion of
one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
18;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 18;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 67 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 18 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
18 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
19;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 19:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 68 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 19 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
19 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellulase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
20;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 20;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 69 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 20 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 20 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,3(4)-beta-glucanse activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
21;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 21;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 70 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 21 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
21 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
22;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 22;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 71 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 22 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 22 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
23;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 23;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 72 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 23 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
23 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
24;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 24;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 73 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 24 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 24 by
substitution, deletion or insertion of
one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
25;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 25:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 74 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 25 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
25 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has dextranase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
26;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 26:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 75 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 26 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
26 by substitution, deletion or insertion of
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one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through 1)
wherein the polypeptide has lysozyme activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
27;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 27;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 76 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 27 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
27 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-tenninal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
28;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 28;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 77 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 28 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
28 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
29;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 29;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 78 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 29 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
29 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has pullulanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
30;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 30;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 79 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 30 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
30 by substitution, deletion or insertion of
one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-alpha-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
31;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 31;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 80 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 31 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
31 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,6-beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
32;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 32:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 81 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 32 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
32 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,6-beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
33;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 33;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 82 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 33 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 33 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has mannan endo-1,4-beta-gluco-mannosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
34;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 34;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 83 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 34 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
34 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has mannan endo-1,6-alpha-gluco-mannosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
35;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 35;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 84 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 35 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 35 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
36;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 36;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 85 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 36 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
36 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
37;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 37;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 86 or the cDNA sequence thereof;
38
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d) a polypeptide derived from SEQ ID NO: 37 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 37 by
substitution, deletion or insertion of
one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
38;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 38:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 87 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 38 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
38 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
39;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 39:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 88 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 39 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
39 by substitution, deletion or insertion of
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one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through 1)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
40;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 40;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 89 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 40 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
40 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-tenninal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
41;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 41;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 90 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 41 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
41 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan 1,4-alpha-maltohydrolase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
42;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 42;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 91 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 42 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
42 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has serine endopeptidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
43;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 43;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 92 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 43 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
43 by substitution, deletion or insertion of
one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glutamyl endopeptidase activity.
In some embodiments, the protein is selected from the group consisting of:
41
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
44;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 44;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 93 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 44 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
44 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has subtilisin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
45;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 45:
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 94 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 45 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
45 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has subtilisin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
46;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 46;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 95 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 46 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 46 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has subtilisin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
47;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 47;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 96 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 47 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
47 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has subtilisin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
48;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 48;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 97 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 48 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 48 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has bacillolysin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
98;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 98;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 151 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 98 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
98 by substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has lytic cellulose monooxygenase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
99;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 99;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 152 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 99 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 99 by
substitution, deletion or insertion of
one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has lytic cellulose monooxygenase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
100;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 100;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 153 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 100 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
100 by substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
101;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 101;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 154 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 101 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
101 by substitution, deletion or insertion
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of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
102;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 102;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 155 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 102 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
102 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
103;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 103;
C) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 156 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 103 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 6 by
substitution, deletion or insertion of
one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has triacylglycerol lipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
104;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 104;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 157 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 104 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
104 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has lysophospholipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
105;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 105;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 158 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 105 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
105 by substitution, deletion or insertion
of one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has pectinesterase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
106;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 106;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 159 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 106 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
106 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has phospholipase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
107;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 107;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 160 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 107 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
107 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has phospholipase C activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
108;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 108;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 59 or the cDNA sequence thereof;
d) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 161 or the cDNA sequence thereof;
e) a polypeptide derived from SEQ ID NO: 108 by substitution, deletion or
insertion of one or more amino
acids;
I) a polypeptide derived from a mature polypeptide of SEQ ID NO:
108 by substitution, deletion or insertion
of one or more amino acids;
g) a polypeptide derived from the polypeptide of any one of a) through f)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
h) a fragment of the polypeptide of any one of a) through g)
wherein the polypeptide has phoshoinositide phospholipase C activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
109;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 109;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 162 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 109 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
109 by substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
110;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 110;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 163 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 110 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 110 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
111;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 111;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 164 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 111 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
111 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
112;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 112;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 165 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 112 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 12 by
substitution, deletion or insertion of
one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
113;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 113;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 166 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 113 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
113 by substitution, deletion or insertion
of one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has alpha-amylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
114;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 114;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 167 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 114 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 114 by
substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellulose activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
115;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 115;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 168 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 115 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
115 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has cellulose activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
116;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 116;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 169 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 116 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
116 by substitution, deletion or insertion
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of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through 1)
wherein the polypeptide has cellulase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
117;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 117;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 170 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 117 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
117 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has inulinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
118;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 118;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 171 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 118 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
118 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
119;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 119;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 172 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 119 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
119 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypcptidc having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
120;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 120;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 173 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 120 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
120 by substitution, deletion or insertion
of one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,4-beta-xylanase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
121;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 121;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 174 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 121 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
121 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
122;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 122;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 175 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 122 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
122 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
123;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 123;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 176 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 123 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 123 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
124;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 124;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 177 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 124 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
124 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
125;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 125;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 178 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 125 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 125 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-polygalacturonase (pectinase) activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
126;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 126;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 179 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 126 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
126 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
127;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 127;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 180 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 127 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 127 by
substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has xylan 1,4-beta-xylosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
128;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 128;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 181 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 128 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
128 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-beta-D-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
129;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 129;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 182 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 129 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
129 by substitution, deletion or insertion
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of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through I)
wherein the polypeptide has alpha-L-arabinofuranosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
130;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 130;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 183 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 130 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
130 by substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-alpha-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
131;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 131;
C) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 184 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 131 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 132 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,3-alpha-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
132;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 132;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 185 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 132 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
132 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has licheninase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypcptidc having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
133;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 133;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 186 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 133 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
133 by substitution, deletion or insertion
of one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has glucan endo-1,6-beta-glucosidase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
134;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 134;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 187 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 134 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
134 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has mannan endo-1,4-p-mannosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
135;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 135;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 188 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 135 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
135 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has 1,4-beta-cellobiosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
136;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 136;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 189 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 136 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 136 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has mannan endo-1,6-alpha-mannosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
137;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 137;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 190 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 137 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
137 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
138;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 138;
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c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 191 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 138 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 138 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endogalactosaminidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
139;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 139;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 192 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 139 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
139 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitosanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
140;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 140;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 193 or the cDNA sequence thereof;
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d) a polypeptide derived from SEQ ID NO: 140 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 140 by
substitution, deletion or insertion
of one or more amino acids;
I) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitosanase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
141;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypcptidc of SEQ ID NO: 141;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 194 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 141 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
141 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has chitinase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
142;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 142;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 195 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 142 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
142 by substitution, deletion or insertion
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of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through 1)
wherein the polypeptide has 1,4-beta-cellobiosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
143;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 143;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 196 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 143 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
143 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has 1,4-beta-cellobiosidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
144;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75. 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 144;
C) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 197 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 144 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 144 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
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g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has serine endopeptidase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
145;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 145;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 198 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ TD NO: 145 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
145 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has subtilisin activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypcptidc having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
146;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 146;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77. 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 199 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 146 by substitution,
deletion or insertion of one or more amino
acids;
c) a polypeptide derived from a mature polypeptide of SEQ ID NO:
146 by substitution, deletion or insertion
of one or more amino acids;
1) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has deacetylase activity.
In some embodiments, the protein is selected from the group consisting of:
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a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
147;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 147;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 200 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 147 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
147 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has deacetylase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
148;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 148;
c) a polypeptide encoded by a polynucleotide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 201 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 148 by substitution,
deletion or insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO:
148 by substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a)
through e) wherein the N- and/or C-terminal
cnd has bccn extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has asparaginase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
149;
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b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 149;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 202 or the cDNA sequence thereof:
d) a polypeptide derived from SEQ ID NO: 149 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 149 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has pectate lyase activity.
In some embodiments, the protein is selected from the group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to SEQ ID NO:
150;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to a mature
polypeptide of SEQ ID NO: 150;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89,
90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to SEQ ID NO: 203 or the cDNA sequence thereof;
d) a polypeptide derived from SEQ ID NO: 150 by substitution, deletion or
insertion of one or more amino
acids;
e) a polypeptide derived from a mature polypeptide of SEQ ID NO: 150 by
substitution, deletion or insertion
of one or more amino acids;
f) a polypeptide derived from the polypeptide of any one of a) through e)
wherein the N- and/or C-terminal
end has been extended by the addition of one or more amino acids; and
g) a fragment of the polypeptide of any one of a) through f)
wherein the polypeptide has endo-1,3(4)-beta-glucanase activity.
In some embodiments, the protein comprises, consists essentially of, or
consists of a wild-type polypeptide. For
example, in some embodiments, the protein comprises, consists essentially of,
or consists of a wild-type polypeptide
having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,
86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % sequence
identity to any one or more of SEQ ID NOs: 1-48
and 98-150 or a mature polypeptide thereof.
In some embodiments, the protein comprises, consists essentially of, or
consists of a variant polypeptide. For
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example, in some embodiments, the protein comprises, consists essentially of,
or consists of a variant polypeptide having
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % sequence identity to
any one or more of SEQ ID NOs: 1-48 and
98-150 or a mature polypeptide thereof.
In some embodiments, the protein comprises, consists essentially of or
consists of a catalytic domain, a binding
module and a linker between said catalytic domain and said binder module.
In some embodiments, the protein comprises two or more catalytic domains.
In some embodiments, the protein comprises two or more binding modules.
In some embodiments, the protein is a fusion protein comprising a first
polypeptide and a second polypeptide,
said second polypeptide being distinct from said first polypeptide, wherein
said first polypeptide is selected from the
group consisting of:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100% sequence identity to any
one or more of SEQ ID NOs: 1-48 and 98-150;
b) a polypcptidc having about/at least 60, 61, 62, 63. 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to a
mature polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75. 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % sequence identity to any one or more of SEQ ID NOs: 49-97 and 151-203 or
the cDNA sequence thereof;
d) a polypeptide derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
e) a polypeptide derived from a mature polypeptide any one of SEQ ID NOs: 1-
48 and 98-150 by
substitution, deletion or insertion of one or more amino acids;
a polypeptide derived from the polypeptide of any one of a) through e) wherein
the N- and/or C-
terminal end has been extended by the addition of one or more amino acids; and
a fragment of the polypeptide of any one of a) through f), and, optionally,
wherein said second polypeptide is selected from the group consisting of:
h) a polypeptide having about/at least 60, 61, 62, 63, 64,
65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any
one or more of SEQ ID NOs: 1-48 and 98-150;
i)a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99
or 100 % sequence identity to a mature
polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150;
j)a polypeptide encoded by a polynucleotide having about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to any one or more of SEQ ID NOs: 49-97 and 151-203or the
cDNA sequence thereof;
k) a polypeptide derived from any one of SEQ ID NOs: 1-48
and 98-150 by substitution, deletion or
insertion of one or more amino acids;
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polypeptide derived from a mature polypeptide any one of SEQ ID NOs: 1-48 and
98-150 by substitution,
deletion or insertion of one or more amino acids;
In) a polypeptide derived from the polypeptide of any one of
II) through 1) wherein the N- and/or C-
terminal end has been extended by the addition of one or more amino acids; and
n) a fragment of the polypeptide of any one of It) through
m).
In preferred embodiments, pmteins of the present disclosure comprise, consist
essentially of or consist of the
amino acid sequence of any one of SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof.
In preferred embodiments, proteins of the present disclosure are encoded by a
polynucleotide that comprises,
consists essentially of or consists of one of SEQ ID NOs: 49-97 and 151-203 or
the cDNA thereof.
In some embodiments, proteins of the present disclosure comprise, consist
essentially of or consist of a
fragment of one of SEQ ID NOs: 1-48 and 98-150 or a mature peptide thereof.
For example, the protein may be a
fragment of any one of SEQ TD NOs: 1-48 and 98-150 or a mature peptide
thereof, said fragment comprising at least 85,
86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 % of the amino acids
found in the original protein.
As noted above, in some embodiments, proteins of the present disclosure
comprise, consist essentially of or
consist of any one of SEQ ID NOs: 1-48 and 98-150 or a mature polypeptide
thereof with an N-tenninal extension of
one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,
15, 16, 17, 18, 19, 20 or more amino acids), a C-
terminal extension of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
more amino acids), one or more substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
more substitutions), one or more insertions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
insertions), and/or one or more deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
deletions). The amino acid changes may be of a minor nature, that is
conservative amino acid substitutions or insertions
that do not significantly affect the folding and/or activity of the protein;
small deletions, typically of 1-30 amino acids;
small amino- or carboxyl-terminal extensions, such as an amino-terminal
methionine residue; a small linker peptide of
up to 20-25 residues; or a small extension that facilitates purification by
changing net charge or another function, such as
a poly-histidine tract, an antigenic epitope or a binding module.
Essential amino acids in a polypeptide can be identified according to
procedures known in the art, such as site-
directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells,
1989, Science 244: 1081-1085). In the
latter technique, single alanine mutations are introduced at every residue in
the molecule, and the resultant molecules are
tested for catalytic activity to identify amino acid residues that are
critical to the activity of the molecule. See also, Hilton
et al., 1996, J. Biol. Chem. 271: 4699-4708. The active site of the enzyme or
other biological interaction can also be
determined by physical analysis of structure, as determined by such techniques
as nuclear magnetic resonance,
crystallography, electron diffraction, or photoaffinity labeling, in
conjunction with mutation of putative contact site amino
acids. See, for example, de Vos et al., 1992, Science 255: 306-312; Smith et
ul., 1992,1 Mol. Biol. 224: 899-904; Wlodaver
et al., 1992, FEBS Lett. 309: 59-64. The identity of essential amino acids can
also be inferred from an alignment with a
related polypeptide, and/or be inferred from sequence homology and conserved
catalytic machinery with a related
polypeptide or within a polypeptide or protein family with
polypeptides/proteins descending from a common ancestor,
typically having similar three-dimensional structures, functions, and
significant sequence similarity. Additionally or
alternatively, protein structure prediction tools can be used for protein
structure modelling to identify essential amino acids
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and/or active sites of polypeptides. See, for example, Jumper et al., 2021,
"Highly accurate protein structure prediction
with AlphaFold". Nature 596: 583-589.
Single or multiple amino acid substitutions, deletions, and/or insertions can
be made and tested using known
methods of mutagenesis, recombination, and/or shuffling, followed by a
relevant screening procedure, such as those
disclosed by Reidhaar-Olson and Sauer, 1988, Science 241: 53-57; Bowie and
Sauer, 1989, Proc. Natl. Acad. S'ci. USA 86:
2152-2156; WO 95/17413; or WO 95/22625. Other methods that can be used include
error-prone PCR, phage display (e.g.,
Lowman et al., 1991, Biochemistry 30: 10832-10837; US 5,223,409; WO 92/06204),
and region-directed mutagenesis
(Derbyshire et al., 1986, Gene 46: 145; Ner et a/., 1988, DNA 7:127).
Mutagenesis/shuffling methods can be combined with high-throughput, automated
screening methods to detect
activity of cloned, mutagenized polypeptides expressed by host cells (Ness et
at., 1999, Nature Biotechnology 17: 893-
896). Mutagenized DNA molecules that encode active polypeptides can be
recovered from the host cells and rapidly
sequenced using standard methods in the art. These methods allow the rapid
determination of the importance of individual
amino acid residues in a polypeptide.
In some embodiments, the protein is a fusion protein (e.g., a fusion protein
comprising a first polypeptide having
a first enzymatic activity and a second polypeptide having a second enzymatic
activity).
In some embodiments, the protein is a hybrid protein.
In some embodiments, the protein is isolated.
In some embodiments, the protein is purified.
It is to be understood that proteins of the present disclosure may be obtained
from microorganisms of any genus.
For purposes of the present disclosure, the term "obtained from" as used
herein in connection with a given source shall
mean that the protein encoded by a polynucleotide is produced by the source or
by a strain in which the polynucleotide of
the disclosure has been inserted. In one aspect, the protein obtained from a
given source is secreted extracellularly.
In some embodiments, the protein is obtained from a Gram-negative bacteria,
such as Campylobacter,
Dicytoglomus (e.g., D. thermophilum), Escherichia (e.g., E. coli),
Flavobacterium, Fusobacterium, Helicobacter,
Ilyobacter, Neisseria, Pseudomonas, Salmonella or Ureaplasma.
In some embodiments, the protein is obtained from a Gram-positive bacteria,
such as Bacillus (e.g., B.
agaradhaerens, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus
brevis, Bacillus circulans, Bacillus clausii,
Bacillus coagulans, B. deramificans, Bacillus jirmus, Bacillus lautus,
Bacillus lentus, Bacillus lichenifOrmis, Bacillus
megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis,
Bacillus thuringiensis), Clostridium,
EnteroCOCCIIS Geobacillu,s, Laciohacilius, Lactococcu,s, Oceanobacillu,s
(e.g., O. barb ara), Staphylococcus,
Streptococcus (e.g., Streptococcus equisimilis, Streptococcus pyogenes,
Streptococcus uberis, and Streptococcus equi
subsp. Zooepidemicus) or Streptomyces (e.g., Streptomyces achromogenes,
Streptomyces avermitilis, Streptomyces
coelicolor, Streptomyces griseus, Streptomyces lividans).
In some embodiments, the protein is obtained from a fungus, such as
Acremonium, Aspergillus (e.g., A. actileatus,
Aspergillus aw am ori Aspergillus foetidus, A spergillus fumigatus,
Aspergillus japonicus,Aspergillus nidulans, Aspergillus
niger, Aspergillus oryzae), Aureobasidium, Bjerkandera (e.g., Bjerkandera
adusta), Ceriporiopsis (e.g., Ceriporiopsis
aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis
pannocinta, Ceriporiopsis rivulosa,
Ceriporiopsis subrufit, Ceriporiopsis subvermispora), Chaetomitun (e.g., C.
erraticum), Chrysosporium (e.g.,
Chrysasporium mops, Chrysasporium keratinophilmn, Chryso,sporium lucknowen,se,
Chryso,sporium merdarium,
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Chrvsosporium pannicola, Chrvsosporium queenslandicum, Chrvsosporium tropicum,
Chiysosporium zonatum),
Coprinus (e.g., Coprinus cinereus), Coriolus (e.g., Coriolus hirsutus),
Cryptococcus, Filibasidium, Fusarium (e.g.,
Fusarium bacirldloides, Fusarium cerealls, Fusarium crook wellen,se, Fusarium
culmorum, Fusarium gram inearum,
Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium
oxysporum, Fusarium reticulatum, Fusarium
roseum, Fusarium sambucinum, Fusarium sarcochroum, F. solani, Fusarium
sporotrichioides, Fusarium sulphureum,
Fusarium torulosum, Fusarium trichothecioides, Fusarium venenatum), Hum/cola
(e.g., Hum/cola insolens, Hum/cola
lanuginosa), Magnaporthe, Microdochium (e.g., M. nivale), Mucor (e.g., illucor
miehei), illyceliophthora (e.g.,
Myceliophthora thermophila), Neocalliinastix, Neurospora (e.g., Neurospora
crassa), Paeciloinyces, Penicilliuin (e.g.,
Penicillium purpurogenum), Phanerochaete (e.g., Phanerochaete chrysosporium),
Phlebia (e.g., Phlebia radiata),
Piromyces, Pleurotus (e.g., Pleurotus eryngii), ,S'chizophyllum,Talaromyces
(e.g., Talaromyces emersonii), Thermoascus
(e.g., T. aurantiactis), Thielavia (e.g., Thielavia terrestris),
Tolypocladium, Trametes (e.g., Trametes villosa, Trametes
veryicolor), or Trichoderma (e.g., T. airoviri de, Trichoderma harzianum,
Trichoderma koningii, Trichoderma
longibrachiatum, Trichoderma reesei, Trichoderma viride).
In some embodiments, the protein is obtained from a yeast, such as Candida,
Hansenula, Kluyveromyces (e.g.,
Kluyveromyces lactis), Pichia, Saccharomyces (e.g., Saccharomyces
carlsbergensis, Saccharomyces cerevisiae,
Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri,
Saccharomyces norbensis,
Saccharomyces oviformis), Schizosaccharomyces, or Yarrowia (e.g., Yarrowia
lipolytica).
It will be understood that for the aforementioned species, the disclosure
encompasses both the perfect and
imperfect states, and other taxonomic equivalents, e.g., anamorphs, regardless
of the species name by which they are
known. Those skilled in the art will readily recognize the identity of
appropriate equivalents.
The proteins may be identified and obtained from other sources including
microorganisms isolated from nature
(e.g., soil, composts, water, etc.) or DNA samples obtained directly from
natural materials (e.g., soil, composts, water, etc.)
using the above-mentioned probes. Techniques for isolating microorganisms and
DNA directly from natural habitats are
well known in the art. A polynucleotide encoding the protein may then be
obtained by similarly screening a gcnomic DNA
or cDNA library of another microorganism or mixed DNA sample. Once a
polynucleotide encoding a protein has been
detected with the probe(s), the polynucleotide can be isolated or cloned by
utilizing techniques that are known to those of
ordinary skill in the art (see, e.g., Davis et al., 2012, Basic Methods in
Molecular Biology, Elsevier).
It is to be understood that proteins of the present disclosure may be produced
using any suitable method(s),
including, but not limited to, shake flask cultivation and large-scale
fermentation (including continuous, batch, fed-batch,
solid-state and/or microcarrier-based fermentation) methods.
The present disclosure extends to methods of producing a protein of the
present disclosure, comprising (a)
cultivating a cell, which in its wild-type form produces the protein, under
conditions conducive for production of the
protein; and optionally, (b) recovering the protein.
In some embodiments, the cell is a Gram-negative bacterial cell, such as
Campylobacter, Escherichia (e.g., E.
colt), Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neisseria,
Pseudomonas, Salmonella or Ureaplasma.
In some embodiments, the cell is a Gram-positive bacterial cell, such as
Bacillus (e.g., Bacillus alkalophilus,
Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus
clausii, Bacillus coagulans, Bacillus firms,
Bacillus lc-tutus, Bacillus lentus, Bacillus licheniformis, Bacillus
megateritiin, Bacillus pumilus, Bacillus
siearoihermophilits, Bacillus subiiiis, Bacillus ihuringiensis), Clostridium,
Enterococcus, Geobacillits, Lactobacillus,
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Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus (e.g.,
Streptococcus equisimilis, Streptococcus pyogenes,
Sh-eptococcus uberis, and Sh-eptococcus equi subsp. Zooepidemicus) or
Streptomyces (e.g., Streptomyces achromogenes,
Streptomyces avermildis, Streplomyce,s coelicolor,Streptomyce,s griseu,s,
Streptomyces
In some embodiments, the cell is a fungal cell, such as Acremonium,
Aspergillus (e.g., Aspergillus awamori,
Aspergillus ,foetidus, Aspergillus ,fumigatus, Aspergillus japonicus,
Aspergillus nidulans, Aspergillus niger, Aspergillus
oryzae), Aureobasidium, Bjerkandera (e.g., Bjerk.andera adusta), Ceriporiopsis
(e.g., Ceriporiopsis aneirina,
Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis pannocinta,
Ceriporiopsis rivulosa, Ceriporiopsis
subrufa, Ceriporiopsis subvermispora), Chrysosporium (e.g., Chrysosporium
mops, Chrysosporium keratinophilum,
Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola,
Chrysosporium queenslandicum,
Chrysosporium tropicum, Chrysosporium zonatum), Coprinus (e.g., Coprinus
cinereus),Coriolus (e.g., Coriolus hirsutus),
Cryptococcus, Filibasidium, Fusarium (e.g., Fusarium bactridioides, Fusarium
cerealis, Fusarium crookwellense,
Fusarium ClilMOTTAM Fir,sarium graminearum, Fir,sarium graminum, Favor/urn
heterosporirm, Fusarium negundi,
Fusarium oxysporum, Fusarium reticulation, Fusarium roseum, Fusarium
sambucinum, Fusarium sarcochroum,
Fusarium sporotrichioides, Fusarium sulphureum, Fusarium torulosum , Fusarium
trichothecioides, Fusarium venenatum),
Humicola (e.g., Humicola insolens, Humicola lanuginosa), Magnaporthe, Mucor
(e.g., Mucor rniehei), Myceliophthora
(e.g., Myceliophthora thermophila)õVeocallimastixõVeurospora (e.g., Neurospora
crassa), Paecilornyces, Penicillium
(e.g., Penicillium purpurogenum), Phanerochaete (e.g., Phanerochaete
chrysosporium), Phlebia (e.g., Phlebia radio/a),
Ptromyces, Pleurotus (e.g., Pleurotus eryngit), Schizophyllum, Talaromyces
(e.g., Talaromyces emersonn), Thermoascus,
Thielavia (e.g., Thielavia terrestris), Tolypocladium, Trametes (e.g.,
Trametes villosa, Trametes versicolor), or
Trichoderma (e.g., Trichoderma harzianum, Trichoderma koningii, Trichoderma
longibrachialum, Trichoderma reesei,
Trichoderma viride).
In some embodiments, the cell is a yeast cell, such as Candida, Hansenula,
Kluyveromyces (e.g., Kluyveromyces
lactis), Pichia, Saccharomyces (e.g., Saccharomyces carlsbergensis,
Saccharomyces cerevisiae, Saccharomyces
diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces
norbensis, Saccharomyces oviform is),
Schizosaccharomyces, or Yarrowia (e.g., Yarrowia hpolytica).
The present disclosure also extends to methods of producing a protein of the
present disclosure, comprising (a)
cultivating a recombinant host cell of the present disclosure under conditions
conducive for production of the protein; and
optionally, (b) recovering the protein.
Cells are cultivated in a nutrient medium suitable for production of the
protein using methods known in the art.
For example, the cell may be cultivated by shake flask cultivation, or small-
scale or large-scale fermentation (including
continuous, batch, fed-batch, or solid-state, and/or microcarrier-based
fermentations) in laboratory or industrial fermentors
in a suitable medium and under conditions allowing the protein to be expressed
and/or isolated. Suitable media are available
from commercial suppliers or may be prepared according to published
compositions (e.g., in catalogues of the American
Type Culture Collection). If the protein is secreted into the nutrient medium,
the protein can be recovered directly from
the medium. If the protein is not secreted, it can be recovered from cell
lysates.
The protein may be detected using methods known in the art that are specific
for the protein, including, but not
limited to, the use of specific antibodies, formation of an enzyme product,
disappearance of an enzyme substrate, or an
assay determining the relative or specific activity of the protein.
The protein may be recovered from the medium using methods known in the art,
including, but not limited to,
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collection, centrifugation, filtration, extraction, spray-drying, evaporation,
or precipitation. In one aspect, a whole
fermentation broth comprising the protein is recovered. In another aspect, a
cell-free fermentation broth comprising the
protein is recovered.
The protein may be purified by a variety of procedures known in the art to
obtain substantially pure proteins
and/or protein fragments (see, e.g., Wingfield, 2015, Current Protocols in
Protein Science; 80(1): 6.1.1-6.1.35; Labrou,
2014, Protein Downstream Processing, 1129: 3-10).
In an alternative aspect, the protein is not recovered.
Proteins of the present disclosure may be used to prevent and/or treat
infestations/infections of/by horticultural
pests at any time __ including prior to planting, at the time of planting,
after planting, prior to germination, after
germination, prior to seedling emergence, at the time of seedling emergence,
after seedling emergence, prior to the
vegetative stage, during the vegetative stage, after the vegetative stage,
prior to the reproductive stage, during the
reproductive stage, after the reproductive stage, prior to flowering, at the
time of flowering, after flowering, prior to
fruiting, at the time of fruiting, after fruiting, prior to ripening, at the
time of ripening, after ripening, prior to harvest, at
the time of harvest, and after harvesting. Accordingly, proteins of the
present disclosure may be formulated for use at any
stage in the horticultural process and by any suitable method of application,
including, but not limited to, on-seed
application, in-furrow application, foliar application, preharvest
application, and postharvest application.
Proteins of the present disclosure may be incorporated into formulations
comprising any suitable carrier,
including, but not limited to, seed-compatible carriers, soil-compatible
carriers, foliar-compatible carriers, preharvest
carriers, and postharvest carriers. Selection of appropriate carrier materials
will depend on the intended application(s)
and the protein(s) to be included in the formulation, as well as any other
components that may be present in and/or added
to the formulation. In some embodiments, the carrier is a liquid, a gel, a
slurry, or a solid. In some embodiments, the
carrier consists essentially of or consists of one or more protein-stabilizing
compounds.
In some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002,
0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007,
0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01,
0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,
0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2,
2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6,
3.7, 3.8, 3.9,4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9,
5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5,
6.6, 6.7, 6.8. 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8,
7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4,
9.5, 9.6, 9.7, 9.8, 9.9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24, 25 % or more protein (i.e., total amount of one or
more proteins of the present disclosure) w/w,
based upon the total weight of the formulation. For example, in some
embodiments, formulations of the present disclosure
comprise about/at least 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004,
0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007,
0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035,
0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45,
0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95,
1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8,
3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4,
5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7,
6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3,
8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6,
9.7, 9.8, 9.9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25
% w/w or more of one or more proteins selected
from the group consisting of:
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a) polypeptides having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 %
sequence identity to one or more of SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof;
b) polypeptides encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any one or more of SEQ ID NOs: 49-
97 and 151-203 or
the cDNA sequence thereof;
c) polypeptides derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
d) polypeptides derived from a mature polypeptide of any one of SEQ ID NOs:
1-48 and 98-150 by
substitution, deletion or insertion of one or more amino acids;
e) polypeptides derived from any one of a) through d) above wherein the N-
and/or C-tenninal end
has been extended by the addition of one or more amino acids;
I) fragments of any one of a) through e) above; and
g) enzymatically active fragments/mutants/variants of any one of
SEQ ID NOs: 1-48 and 98-150 or
mature polypeptides thereof.
In some embodiments, formulations of the present disclosure comprise about
0.0001 to about 40% protein (i.e.,
total amount of one or more proteins of the present disclosure) w/w, based
upon the total weight of the formulation. For
example, in some embodiments, formulations of the present disclosure comprise
about 0.001, 0.0015. 0.002, 0.0025, 0.003,
0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008,
0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025,
0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085,
0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4,
0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3,
1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1,
4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5 % w/w to about
0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2,
2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6,
3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9,
5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5,
6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8,
7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4,
9.5, 9.6, 9.7, 9.8, 9.9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 21, 22, 23, 24 or 25 % w/w (e.g., about 0.0001 to about 1 (Yow/w) of
one or more pH control components selected
from the group consisting of proteins selected from the group consisting of:
a) polypeptides having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 %
sequence identity to one or more of SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof;
b) polypeptides encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any one or more of SEQ ID NOs: 49-
97 and 151-203 or
the cDNA sequence thereof;
c) polypeptides derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
d) polypeptides derived from a mature polypeptide of any one of SEQ ID NOs:
1-48 and 98-150 by
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substitution, deletion or insertion of one or more amino acids;
e) polypeptides derived from any one of a) through d) above wherein the N-
and/or C-terminal end
has been extended by the addition of one or more amino acids;
f) fragments of any one of a) through e) above; and
g) enzymatically active fragments/mutants/variants of any one of SEQ ID
NOs: 1-48 and 98-150 or
mature polypeptides thereof.
The present disclosure extends to granules/particles comprising one or more
proteins of the disclosure. In an
embodiment, the granule comprises a core, and optionally one or more coatings
(outer layers) surrounding the core.
The core may have a diameter, measured as equivalent spherical diameter
(volume based average particle size),
of 20-2000 um, particularly 50-1500 um, 100-1500 um or 250-1200 um. The core
diameter, measured as equivalent
spherical diameter, can be determined using laser diffraction, such as using a
Malvern Mastersizer and/or the method
described under IS013320 (2020).
in an embodiment, the core comprises one or more proteins of the present
disclosure.
The core may include additional materials such as fillers, fiber materials
(cellulose or synthetic fibers), stabilizing
agents, solubilizing agents, suspension agcnts, viscosity regulating agents,
light spheres, plasticizers, salts, lubricants and
fragrances.
The core may include a binder, such as synthetic polymer, wax, fat, or
carbohydrate.
The core may include a salt of a multivalent cation, a reducing agent, an
antioxidant, a peroxide decomposing
catalyst and/or an acidic buffer component, typically as a homogenous blend.
The core may include an inert particle with the protein absorbed into it, or
applied onto the surface, e.g., by fluid
bed coating.
The core may have a diameter of 20-2000 um, particularly 50-1500 um, 100-1500
um or 250-1200 um.
The core may be surrounded by at least one coating, e.g., to improve the
storage stability, to reduce dust formation
during handling, or for coloring the granule. The optional coating(s) may
include a salt coating, or other suitable coating
materials, such as polyethylene glycol (PEG), methyl hydro.xy-propyl cellulose
(MI-IPC) and polyvinyl alcohol (PVA).
The coating may be applied in an amount of at least 0.1% by weight of the
core, e.g., at least 0.5%, at least 1%,
at least 5%, at least 10%, or at least 15%. The amount may be at most 100%,
70%, 50%, 40% or 30%.
The coating is preferably at least 0.1 gm thick, particularly at least 0.5 gm,
at least 1 gm or at least 5 gm. In some
embodiments, the thickness of the coating is below 100 gm, such as below 60
gm, or below 40 gm.
The coating should encapsulate the core mnt by forming a substantially
continuous layer. A substantially
continuous layer is to be understood as a coating having few or no holes, so
that the core unit has few or no uncoated areas.
The layer or coating should, in particular, be homogeneous in thickness.
The coating can further contain other materials as known in the art, e.g.,
fillers, antisticking agents, pigments,
dyes, plasticizers and/or binders, such as titanium dioxide, kaolin, calcium
carbonate or tale.
A salt coating may comprise at least 60% by weight of a salt, e.g., at least
65%, at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95% or at least 99% by weight.
To provide acceptable protection, the salt coating is preferably at least 0.1
gm thick, e.g., at least 0.5 gm, at least
1 gm, at least 2 gm, at least 4 gm, at least 5 gm, or at least 8 gm. In a
particular embodiment, the thickness of the salt
coating is below 100 gm, such as below 60 am, or below 40 am.
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The salt may be added from a salt solution where the salt is completely
dissolved or from a salt suspension wherein
the fine particles are less than 50 gm, such as less than 10 gm or less than 5
gm.
The salt coating may comprise a single salt or a mixture of two or more salts.
The salt may be water soluble, in
particular, having a solubility at least 0.1 gin 100 g of water at 20 C,
preferably at least 0.5 g per 100 g water, e.g., at least
1 g per 100 g water, e.g., at least 5 g per 100 g water.
The salt may be an inorganic salt, e.g., salts of sulfate, sulfite, phosphate,
phosphonate, nitrate, chloride or
carbonate or salts of simple organic acids (less than 10 carbon atoms, e.g., 6
or less carbon atoms) such as citrate, malonate
or acetate. Examples of cations in these salts are alkali or earth alkali
metal ions, the ammonium ion or metal ions of the
first transition series, such as sodium, potassium, magnesium, calcium, zinc
or aluminum. Examples of anions include
chloride, bromide, iodide, sulfate, sulfite, bisulfite, thiosulfate,
phosphate, monobasic phosphate, dibasic phosphate,
hypophosphite, dihydrogen pyrophosphate, tetraborate, borate, carbonate,
bicarbonate, metasilicate, citrate, malate,
maleate, malonate, succinate, lactate, formate, acetate, butyrate, propionate,
benzoate, tartrate, ascorbate or gluconate. In
particular, alkali- or earth alkali metal salts of sulfate, sulfite,
phosphate, phosphonate, nitrate, chloride or carbonate or
salts of simple organic acids such as citrate, malonate or acetate may be
used.
The salt in the coating may have a constant humidity at 20 C above 60%,
particularly above 70%, above 80% or
above 85%, or it may be another hydrate form of such a salt (e.g., anhydrate).
The salt coating may be as described in WO
00/01793 or WO 2006/034710.
Specific examples of suitable salts are NaCl (CH2orc=76%), Na2CO3 (CH20-
c=92%), NaNO3 (CH2o-c=73N,
Na2HPO4 (CH200c=95%), Na3PO4 (CH250c=92`)/o), NI-14C1 (CHnoc = 79.5%), (NI-
14)2HPO4 (CH?vc = 93,0%), NH4H2PO4
(CH200c = 93.1%), (N1-14)2SO4 (CH200c=81.1%), KCl (CH2occ=85%), K2HPO4
(CH200c=92%), KH2PO4 (CH200c=96.5 A),
KNO3 (CH200c=93.5%), Na2SO4 (CH200c=93%), K2S01 (CH200c=98%), KHS01
(CH200c=86%), MgSO4 (CH200c=90 /0),
ZnSO4 (CH200c=90%) and sodium citrate (CH250c=86%). Other examples include
NaH2PO4, (NI-14)H2PO4, CuSO4,
Mg(NO3)2 and magnesium acetate.
The salt may be in anhydrous form, or it may be a hydrated salt, i.e., a
crystalline salt hydrate with bound water(s)
of crystallization, such as described in WO 99/32595. Specific examples
include anhydrous sodium sulfate (Na2SO4),
anhydrous magnesium sulfate (MgS01), magnesium sulfate heptahydrate
(MgS017H20), zinc sulfate heptahydrate
(ZnSO4.7H20), sodium phosphate dibasic heptahydrate (Na2HPO4.7H20), magnesium
nitrate hexahydrate
(Mg(N01)2(6H20)), sodium citrate dihydrate and magnesium acetate tetrahydrate.
Preferably the salt is applied as a solution of the salt, e.g., using a fluid
bed.
The coating materials can be waxy coating materials and film-forming coating
materials. Examples of waxy
coating materials are poly(ethylene oxide) products (polyethyleneglycol, PEG)
with mean molar weights of 1000 to 20000;
ethoxylated nonylphenols having from 16 to 50 ethylene oxide units;
ethoxylated fatty alcohols in which the alcohol
contains from 12 to 20 carbon atoms and in which there are 15 to 80 ethylene
oxide units; fatty alcohols; fatty acids; and
mono- and di- and triglycerides of fatty acids. Examples of film-forming
coating materials suitable for application by fluid
bed techniques are given in GB 1483591.
The granule may optionally have one or more additional coatings. Examples of
suitable coating materials are
polyethylene glycol (PEG), methyl hydroxy-propyl cellulose (MTIPC) and
polyvinyl alcohol (PVA). Examples of enzyme
granules with multiple coatings are described in WO 93/07263 and WO 97/23606.
The core can be prepared by granulating a blend of the ingredients, e.g., by a
method comprising granulation
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techniques such as crystallization, precipitation, pan-coating, fluid bed
coating, fluid bed agglomeration, rotary atomization,
extrusion, prilling, spheronization, size reduction methods, drum granulation,
and/or high shear granulation.
Methods for preparing the core can be found in the Handbook of Powder
Technology; Particle size enlargement
by C. E. Capes; Vol. 1; 1980; Elsevier. Preparation methods include known feed
and granule formulation technologies,
e.g.,
(a) Spray dried products, wherein a liquid protein-containing solution is
atomized in a spray drying tower to form
small droplets which during their way down the drying tower diy to form a
protein-containing particulate material. Veiy
small particles can be produced this way (Michael S. Showell (editor);
Powdered detergents; Surfactant Science Series;
1998; Vol. 71; pages 140-142; Marcel Dekker).
(b) Layered products, wherein the protein is coated as a layer around a pre-
formed inert core particle, wherein a
protein-containing solution is atomized, typically in a fluid bed apparatus
wherein the pre-formed core particles are
fluidized, and the protein-containing solution adheres to the core particles
and dries up to leave a layer of dry protein on
the surface of the core particle. Particles of a desired size can be obtained
this way if a useful core particle of the desired
size can be found. This type of product is described in, e.g., WO 97/23606.
(c) Absorbed core particles, wherein rather than coating the protein as a
layer around the core, the protein is
absorbed onto and/or into the surface of the core. Such a process is described
in WO 97/39116.
(d) Extrusion or pelletized products, wherein a protein-containing paste is
pressed to pellets or under pressure is
extruded through a small opening and cut into particles which are subsequently
dried. Such particles usually have a
considerable size because of the material in which the extrusion opening is
made (usually a plate with bore holes) sets a
limit on the allowable pressure drop over the extrusion opening. Also, very
high extrusion pressures when using a small
opening increase heat generation in the protein paste, which is harmful to the
protein (Michael S. Showell (editor);
Powdered detergents; Surfactant Science Series; 1998; Vol. 71; pages 140-142;
Marcel Dekker).
(e) Prilled products, wherein a protein-containing powder is suspended in
molten wax and the suspension is
sprayed, e.g., through a rotating disk atomizer, into a cooling chamber where
the droplets quickly solidify (Michael S.
Showell (editor); Powdered detergents; Surfactant Science Series; 1998; Vol.
71; pages 140-142; Marcel Dekker). The
product obtained is one wherein the protein is uniformly distributed
throughout an inert material instead of being
concentrated on its surface. US 4,016,040 and US 4,713,245 describe this
technique.
(f) Mixer granulation products, wherein a protein-containing liquid is added
to a dry powder composition of
conventional granulating components. The liquid and the powder in a suitable
proportion are mixed and as the moisture of
the liquid is absorbed in the dry powder, the components of the dry powder
will start to adhere and agglomerate and
particles will build up, forming granulates comprising the protein. Such a
process is described in US 4,106,991, EP 170360,
EP 304332, EP 304331, WO 90/09440 and WO 90/09428. In a particular aspect of
this process, various high-shear mixers
can bc uscd as granulators. Granulates consisting of protein, fillers and
bindcrs ctc. arc mixed with cellulose fibers to
reinforce the particles to produce a so-called T-granulate. Reinforced
particles, are more robust, and release less enzymatic
dust.
(g) Size reduction, wherein the cores are produced by milling or crushing of
larger particles, pellets, tablets,
briquettes etc. containing the protein. The wanted core particle fraction is
obtained by sieving the milled or crushed product.
Over and undersized particles can be recycled. Size reduction is described in
Martin Rhodes (editor); Principles of Powder
Technology; 1990; Chapter 10; John Wiley & Sons.
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(h) Fluid bed granulation. Fluid bed granulation involves suspending
particulates in an air stream and spraying a
liquid onto the fluidized particles via nozzles. Particles hit by spray
droplets get wetted and become tacky. The tacky
particles collide with other particles and adhere to them to form a granule.
(i) The cores may be subjected to drying, such as in a fluid bed drier. Other
known methods for diying granules
in the feed or enzyme industry can be used by the skilled person. The drying
preferably takes place at a product temperature
of from 25 to 90 C. For some proteins, it is important the cores comprising
the protein contain a low amount of water
before coating with the salt. If water sensitive proteins are coated with a
salt before excessive water is removed, the
excessive water will be trapped within the core and may affect the activity of
the protein negatively. After drying, the cores
preferably contain 0.1-10% w/w water.
Non-dusting granulates may be produced, e.g., as disclosed in US 4,106,991 and
US 4,661,452 and may optionally
be coated by methods known in the art.
The granulate may further comprise one or more additional enzymes, e.g.,
hydrolase, isomerase, ligase, lyase,
oxidoreductase, and transferase. The one or more additional enzymes are
preferably selected from the group consisting of
acetylxylan esterase, acylglycerol lipase, amylase, alpha-amylase, beta-
amylase, arabinofuranosidase, cellobiohydrolases,
cellulase, feruloyl esterase, galactanasc, alpha-galactosidasc, beta-
galactosidasc, beta-glucanase, beta-glucosidasc,
lysophospholipase, lysozyme, alpha-mannosidase, beta-mannosidase (mannanase),
phytase, phospholipase Al,
phospholipase A2, phospholipase D, protease, pullulanase, pectin esterase,
triacylglycerol lipase, xylanase, beta-xylosidase
or any combination thereof. Each enzyme will then be present in more granules
securing a more uniform distribution of
the enzymes, and also reduces the physical segregation of different enzymes
due to different particle sizes. Methods for
producing multi-enzyme co-granulates is disclosed in the ip.com disclosure
IPCOM000200739D. Another example of
formulation of proteins using co-granulates is disclosed in WO 2013/188331.
The present disclosure also relates to protected proteins prepared according
to the method(s) disclosed in EP
238216.
The present disclosure also extends to liquid formulations comprising one or
more proteins of the disclosure. In
some embodiments, the fornmlation comprises one or more enzyme stabilizers,
one or more pH control components, one
or more rain fasteners, and, optionally, one or more preservatives.
Formulations of the present disclosure may comprise any suitable enzyme
stabilizer(s), including, but not limited
to, sugars (e.g., monosaccharides, such as fructose. galactose and glucose;
disaccharides, such as lactose, maltose and
sucrose; oligosaccharides, such as maltodextrins; and polysaccharides, such as
celluloses and starches), polyols (e.g., sugar
alcohols, such as arabitol, erythritol, ethylene glycol, glycerol, mannitol,
sorbitol and xylitol; and polymeric polyols, such
as polyethylene glycols and polypropylene glycols), polyvinyl alcohols, lactic
acid, lactic acid derivatives, boric acids,
boric acid derivatives (e.g., aromatic borate esters and phenyl boronic acid
derivatives, such as 4-formylphenyl boronic
acid), and reversible protcasc inhibitors. See generally, e.g., WO
2007/113241; WO 01/04279; WO 2013/004636; WO
95/02046; WO 2009/118375; WO 2020/115179; WO 96/41859; WO 2007/025549; WO
96/23062; WO 2018/130654;
WO 96/22366; WO 92/17571; WO 2017/044473; WO 2017/044545, WO 2017/116837, WO
2017/116846, WO
2017/210163, WO 2017/210166, WO 2018/118740, WO 2018/175681, WO 2018/183491,
WO 2018/218008, WO
2018/218016; WO 2018/218035. It is to be understood that certain enzyme
stabilizers may impart other beneficial
properties to formulations of the present disclosure, such as enhanced
flowability and/or improved adhesion to a plant
surface.
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in some embodiments, formulations of the present disclosure comprise about/at
least 5, 10, 15, 20, 25, 30, 35, 40,
45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 % or more polyol (i.e., total
amount of one or more polyols) w/w, based upon the
total weight of the formulation. For example, in some embodiments,
formulations of the present disclosure comprise
about/at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80,
85, 90, 95 % w/w or more of one or more polyols
selected from the group consisting of glycerol, sorbitol, propylene glycol
(MPG), ethylene glycol, diethylene glycol,
triethylene glycol, 1,2-propylene glycol or 1,3-propylene glycol, dipropylene
glycol, polyethylene glycols (PEG) having
an average molecular weight below about 600, and polypropylene glycols (PPG)
having an average molecular weight
below about 600, preferably glycerol, sorbitol and/or propylene glycol (MPG).
In some embodiments, formulations of the present disclosure comprise about 5
to about 95 % polyol (i.e., total
amount of one or more polyols) w/w, based upon the total weight of the
formulation. For example, in some embodiments,
formulations of the present disclosure comprise about 5, 10, 15, 20,25, 30,
40, 45 or 50 % w/w to about 50, 55, 60, 65, 70,
75, 80, 85, 90 or 95 % w/w (e.g., about 20 to about 40 % w/w) of one or more
polyols selected from the group consisting
of glycerol, sorbitol, propylene glycol (MPG), ethylene glycol, diethylene
glycol, triethylene glycol, 1,2-propylene glycol
or 1,3-propylene glycol, dipropylene glycol, polyethylene glycols (PEG) having
an average molecular weight below about
600, and polypropylene glycols (PPG) having an average molecular wcight below
about 600, preferably glycerol, sorbitol
and/or propylene glycol (MPG).
Formulations of the present disclosure may comprise any suitable pH control
component(s), including, but not
limited to, acetate, carbonate, citrate, phosphate and other salts capable of
buffering the formulation at a desired pH and
having an aqueous solubility of more than 1% w/w. Preferred pH control
components in some embodiments are phosphate
buffers containing the ionic species HP042- and H3PO4-.
A pH control component may be a single ionic species, that can maintain a
constant pH but only provide a
buffering effect towards either acidification or basification. An example of
such, is HP042- which can ensure an alkaline
pH (of approximately 9) and provide a buffering effect against acidification.
This may be beneficial in an agricultural
setting to keep the pH constant at an alkaline pH, as most environmental
factors will cause acidification of a droplet and
deposit.
In preferred embodiments, the pH control component does not significantly
change pH (+/- 0.5 pH units) or
change in a desired direction upon drying when the solvent evaporates from a
droplet on a plant surface. Some buffers
will, upon drying, change pH as a result of differences in solubility of the
buffer components. As an example, the pH of a
sodium phosphate buffer constituting of Na2HPO4 and NaH2PO4 can reduce to pH 4
or lower upon drying since the
dibasic form (Na3HPO4) will crystallize to a larger degree. On the contrary,
the pH of a potassium phosphate buffer
constituting of 1(31-1PO4 and KH2PO4 will approach pH 9 upon diying since the
monobasic form (KH2PO4) has the lowest
solubility (Sarciaux 1999).
A pH control component is most effective (highest buffer capacity) whcn thc
pKa is close to the desired pH of
the composition. This will reduce the amount of buffer needed to maintain a
desired pH. In an embodiment, the buffer
includes salts having a neutral/alkaline pKa, such as a pKa in the range of
6.5 to 10.
As a rule of thumb, a pH control component can be used to control the pH of a
solution at a pH +/- 1 pH-unit
from its pKa value. For example, pH control components with a pKa value above
6.5 are useful for controlling the pH at
7.5 or above. Examples of suitable pH control components include, but are not
limited to sodium or potassium phosphate
(pKal 2.12, pKa2 7.21, pKa3 12.67), sodium or potassium carbonate (pKal 6.37,
pKa2 10.32), 2-amino-2-
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(hydroxymethyl)-1,3-propanediol (TR1S) (pKa 8.1), IBis(2-
hydroxyethyl)aminoJacetic acid (Bicine) (pKa 8.35), N-
Itris(hydroxymethyl)methyllglycine (Tricine) (pKa 8.15), 4-(2-hydroxyethyl)-1-
piperazineethanesulfonic acid (HEPES)
(pKai 3.0, pKa2 7.5), N-l_tris(hydroxymethyl)methyll- 2-aminoethanesulfonic
acid (TES) (pKa 7.55), 3-(N-
morpholino)propanesulfonic acid (MOPS) (pKa 7.2),
tris(hydroxymethyl)methylaminolpropanesulfonic acid (TAPS)
(pKa 8.44), N-Itris(hydroxymethyl)methy11-3-amino-2-hydroxypropanesulfonic
acid (TAPSO) (pKa 7.6), glycylglycine
(pKal 3.14 pKa2 8.17), 2-(N-cyclohexylamino)ethanesulfonic acid (CHES) (pKa
9.3), sodium or potassium borate (pKat
9.24, pKa2 12.4, pKa3 13.3), 2-amino-2-methyl-1,3-pmpanediol (ammediol) (pKa
8.8), triethanol amine (pKa 7.74), 2-
amino-2-methyl-1-propanol (pKa 9.7), glycine (pKal 2.34, pKa2 9.6), histidine
(pKal 1.82, pKa2 6.00, pKa3 9.17), and
other amino acid buffers.
Non-preferred pH control components include, but are not limited to, pH
control components with an
unfavorable pKa (e.g., pKa <6.5 for an enzyme that requires an alkaline pH or
pKa >7.5 for an enzyme that requires an
acidic pH), volatile pH control component, pH control component that display
significant phytotoxicity (this may
sometimes include the above-mentioned "suitable" pH control components, as
phytotoxicity is depended on buffer
concentration, pH and target crop), and pH control components that are
unwanted in the environment and therefore
regulated by authorities (this may sometimes include the above-mentioned
"suitable" pH control component, as
regulations vary throughout the world).
In some embodiments, pH control components may be used to provide formulations
of the present disclosure
that maintain an alkaline pH. For example, in some preferred embodiments,
formulations of the present disclosure
comprise one or more pH control components selected to provide a composition
having an alkaline pH, preferably at
least 7.5, more preferably between 7.5 and 10, most preferably between 8 and
9.5. Thus, in some embodiments,
formulations of the present disclosure comprise a pH control component, such
as a buffer, where an 1% w/w aqueous
solution of the pH control component (buffer) has an alkaline pH (e.g., above
7.5, preferably above 8, and below 10,
preferably below 9.5).
In some embodiments, pH control components may be used to provide formulations
of the present disclosure
that maintain a neutral pH. For example, in some preferred embodiments,
formulations of the present disclosure
comprise one or more pH control components selected to provide a composition
having a neutral pH, preferably between
6.5 and 7.5, more preferably between 6.75 and 7.25, most preferably about 7.
Thus, in some embodiments, formulations
of the present disclosure comprise a pH control component, such as a buffer,
where an 1% w/w aqueous solution of the
pH control component (buffer) has a neutral pH (e.g., above 6.5 and below 7.5,
preferably above 6.75 and below 7.25,
more preferably about 7).
In some embodiments, pH control components may be used to provide formulations
of the present disclosure
that maintain an acidic pH. For example, in some preferred embodiments,
formulations of the present disclosure
comprise one or more pH control components selected to provide a composition
having an acidic pH, preferably below
6.5, more preferably between 4 and 6.5, most preferably between 4.5 and 6.
Thus, in some embodiments, formulations of
the present disclosure comprise a pH control component, such as a buffer,
where an 1% w/w aqueous solution of the pH
control component (buffer) has an acidic pH (e.g., below 6.5, preferably below
6, and above 4, preferably above 4.5).
In some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002,
0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007,
0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01,
0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,
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0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2,
2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6,
3.7, 3.8, 3.9,4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,
4.9, 5 % or more pH control component (i.e., total amount of one or more pH
control components) w/w, based upon the
total weight of the formulation. For example, in some embodiments,
formulations of the present disclosure comprise
about/at least 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045,
0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075,
0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04,
0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075,
0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5,
0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1,
1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7,
2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4,
4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5 % w/w or more of one or more pH
control components selected from the group
consisting of acetate, citrate, carbonate and phosphate buffers, preferably
sodium acetate, sodium citrate, sodium
carbonate and/or potassium phosphate buffers.
In some embodiments, formulations of the present disclosure comprise about
0.001 to about 10 % pH control
component (i.e., total of one or more pH control components) w/w, based upon
the total weight of the formulation. For
example, in some embodiments, formulations of the present disclosure comprise
about 0.001, 0.0015, 0.002, 0.0025,
0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075,
0.008, 0.0085, 0.009, 0.0095, 0.01, 0.015,
0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075,
0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25,
0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or
1 % w/w to about 0.25, 0.3, 0.35, 0.4, 0.45, 0.5,
0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5,
1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7,
2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5 % w/w (e.g., about 0.01
to about 1 % w/w) of one or more pH control components selected from the group
consisting of acetate, carbonate,
citrate and phosphate buffers, preferably sodium acetate, sodium carbonate,
sodium citrate and/or potassium phosphate
buffers.
Formulations of the present disclosure may comprise any suitable rain
fastener(s), including, but not limited to,
organo-modified siloxanes (organosiloxanes), such as organo-modified
trisiloxanes (e.g., polyether-modified trisiloxanes,
such as polyalkyleneoxide-modified heptamethyltrisiloxane) and organo-
modifiedpolysiloxanes (e.g., polyether-modified
polysiloxanes,). See generally, e.g., EPO 0245970; US 5496568; WO 2008/144024;
WO 2009/135049; WO 2011/126832;
WO 2017/083049; WO 2020/225276; WO 2021/055316; WO 2022/096688; WO
2022/096691; WO 2022/096692; WO
2022/096693; WO 2022/096694; WO/2022/096695; WO 2022/096696.
In some embodiments, formulations of the present disclosure comprise one or
more organo-modified siloxanes
haying the general molecular structure of Formula 1:
R13SiO[R12SiO1A1R1R2SiOleSiR13
in which
RI- represents identical or different from each other hydrocathon substituents
of 1-10 carbons or hydrogen, preferred are
methyl, ethyl, propyl and phenyl substituents, particularly preferred are
methyl substituents;
R2 represents identical or different from each other polyether substituents of
the general Formula II:
-R30[CH2CH201c[CH2CH(CH3)01D[CHRICHR401ER5
wherein
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R3 represents identical or different from each other hydrocarbon moieties of 1-
8 carbons, which
optionally is interrupted by oxygen atoms. Preferred is linear hydrocarbons of
2-4 carbons, particularly
preferred is -CH2-CH2-CH2-
represents identical or different from each other hydrocathon substituents of
1-12 carbons or
hydrogen, preferred is methyl, ethyl, phenyl or hydrogen substituents
R' represents identical or different from each other hydrocarbon substituents
of 1-16 carbons, which
optionally contains urethane, carbonyl or carboxylic acid functionality, or
hydrogen.
Methyl or hydrogen substituents are preferred, with hydrogen being most
preferred. A is 0-200, preferably 0-1, more
preferably 0. B is 0-200, preferably 0.5-2, more preferably 1. In preferred
embodiments, A+B > 0. C is 0-60, preferably
1-15. D is 0-60, preferably 0-10. E is 0-20, preferably 0-10, more preferably
0. In preferred embodiments, C+D+E > 0.
A trisiloxane may be defined as a molecule of the general Formula I with A = 0
and B = 1, whereas a
polysiloxane is a molecule of the general formula I with A + B >1 and A >1.
Examples of commercially available organo-modified siloxanes include, but are
not limited to BIOSPREAD
surfactants (Grosafc Chemicals Ltd., New Zealand); BREAK-THRUO surfactants
(Evonik Operations Gmbh, Essen,
Germany), such as BREAK-THRU AF 5503, BREAK-THRU AF 9902, BREAK-THRU AF
9903, BREAK-
THRUM OE 440, BREAK-THRU OE 444, BREAK-THRU OE 446, BREAK-THRU S 200, BREAK-
THRU S
233, BREAK-THRU S 240, BREAK-THRU S 255, BREAK-THRU S 279, BREAK-THRU S
301, BREAK-
THRU SD 260, and BREAK-THRU UNION; BYK surfactants (BYK-Chemie GmbH, Wesel,
Germany), such as
BYK -348; ECOSPREADO surfactants (Grosafe Chemicals Ltd., New Zealand); HI-
WETT surfactants (Loveland
Products, Inc., Greeley, CO, USA); and SILWETTm surfactants (Momentive, Inc.,
Waterford, NY, USA), such as
SILWETTm L-77, SILWETTm HS-312, SILWETTm 408, SILWETTm 618, SILWETTm 625,
SILWETTm 636, SILWETTm
641, SILWETTm 806, SILWETTm DA-40, SILWETTm DRS-60, SILWETTm ECO, SILWETTm
FUSION, SILWETTm HS
312, SILWETTm HS 604, SILWETTm HSEC, SILWETTm LF, SILWETTm OC, and SILWETTm
STIK 2.
In some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002,
0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007,
0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01,
0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,
0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2,
2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6,
3.7, 3.8, 3.9,4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,
4.9, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,
24, 25, 30, 35, 40, 45, 50 % or more rain fastener
(i.e., total amount of one or more rain fasteners) w/w, based upon the total
weight of the formulation. For example, in
some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002, 0.0025, 0.003,
0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065. 0.007, 0.0075, 0.008,
0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02,
0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3,
0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1,
1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2,
2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8,
3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6,7,
8, 9, 10 % w/w or more of one or more rain fasteners selected from the group
consisting of organo-modified siloxanes,
preferably organo-modified trisiloxanes and organo-modified polysiloxanes,
more preferably organo-modified
trisiloxanes and poly siloxanes comprising one or more polyether groups, most
preferably trisiloxane (poly)ethoxylates
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and polysiloxane (poly)ethoxylates.
In some embodiments, formulations of the present disclosure comprise about
0.001 to about 50 % rain fastener
(i.e., total amount of one or more rain fasteners) w/w, based upon the total
weight of the formulation. For example, in
some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002, 0.0025, 0.003,
0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008,
0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02,
0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3,
0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or 1 %
w/w to about 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8,
0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2,
2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4,
3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5,6,
7, 8,9 or 10 % w/w of one or more rain fasteners
selected from the group consisting of organo-modified siloxanes, preferably
organo-modified trisiloxanes and organo-
modified polysiloxanes, more preferably organo-modified trisiloxanes and
polysiloxanes comprising one or more
polyether groups, most preferably trisiloxane (poly)ethoxylates and
polysiloxane (poly)ethoxylates.
Formulations of the present disclosure may comprise any suitable
preservative(s), including, but not limited to,
potassium benzoate, potassium sorbate, sodium benzoate and sodium sorbate.
In some embodiments, formulations of the present disclosure comprise about/at
least 0.001, 0.0015, 0.002,
0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007,
0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01,
0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07,
0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2,
0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9,
0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2,
2.1, 2.2, 2.3, 2.4, 2.5, 2.6. 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6,
3.7, 3.8, 3.9,4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,
4.9, 5, 6, 7, 8, 9, 10 % or more preservative (i.e., total amount of one or
more preservatives) w/w, based upon the total
weight of the formulation. For example, in some embodiments, formulations of
the present disclosure comprise about/at
least 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005,
0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008,
0.0085, 0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045,
0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08,
0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55,
0.6. 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2,
1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9,2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8,
2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4,41,
4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5 % w/w or more of one or more
preservatives selected from the group consisting of
potassium benzoate, potassium sorbate, sodium benzoate and sodium sorbate,
preferably potassium sorbate and/or
sodium benzoate.
In some embodiments, formulations of the present disclosure comprise about
0.001 to about 10 % preservative
(i.e., total amount of one or more preservatives) w/w, based upon the total
weight of the formulation. For example, in
some embodiments, formulations of the present disclosure comprise about 0.001,
0.0015, 0.002, 0.0025, 0.003, 0.0035,
0.004, 0.0045, 0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085,
0.009, 0.0095, 0.01, 0.015, 0.02, 0.025, 0.03,
0.035, 0.04, 0.045, 0.05, 0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09,
0.095, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45,
0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95 or 1 % w/w to about 0.5,
0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95,
1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5,
2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8,
3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5 % w/w of one or more
preservatives selected from the group
consisting of potassium benzoate, potassium sorbate, sodium benzoate and
sodium sorbate, preferably potassium sorbate
and/or sodium benzoate.
Proteins of the present disclosure may be incorporated into formulations
comprising myriad components,
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including, but not limited to, adhesives (stickers), chemical actives,
dispersants (spreaders), drying agents. emulsifiers,
microbes, nutrients, pest attractants and feeding stimulants, pH control
components, postharvest treatments, rhealogical
agents, safeners, UV protectants and/or wetting agents.
Examples of adhesives (stickers) that may be included in formulations of the
present disclosure include, but not
are not limited to, disaccharides (e.g. maltose, sucrose, trehalose), gums
(e.g., cellulose gum, guar gum, gum arabic, gum
combretum, xantham gum), maltodextrins (e.g., maltodextrins having a DEV of
about 10 to about 20), monosaccharides,
oils (e.g., mineral oil, olive oil, peanut oil, soybean oil and/or sunflower
oil), and oligosaccharides. See generally, e.g.,
POWERBLOXTM (Dow, Midland, MI, USA), such as POWERBLOXTM ADJ-65 and
POWERBLOXTM ADJ-65; EPO
0245970; US 5496568; WO 2008/144024; WO 2009/135049; WO 2011/126832; WO
2017/083049; WO 2020/225276;
WO 2021/055316; WO 2022/096688: WO 2022/096691; WO 2022/096692; WO
2022/096693: WO 2022/096694;
WO/2022/096695; WO 2022/096696.
Examples of chemical actives that may be included in formulations of the
present disclosure include, but not are
not limited to, acaracides and miticides (e.g., carvacrol, sanguinanne,
azobenzene, benzoximate, benzyl benzoate,
bromopropylate, chlorbenside, chlorfenethol, chlorfenson, chlorfensulphide,
chlorobenzilate, chloropropylate,
cyflumetofen, DDT, dicofol, diphenyl sulfonc, dofcnapyn, fcnson, fcntrifanil,
fluorbenside, gcnit, hexachlorophene,
phenproxide, proclonol, tetradifon, tetrasul, benomyl, carbanolate, carbaiyl,
carbofuran, methiocarb, metolcarb,
promacyl, propoxur, aldicarb, butocarboxim, oxamyl, thiocarboxime, thiofanox,
bifenazate, binapacryl, dinex,
dinobuton, dinocap-4, dinocap-6, dinocton, dinopenton, dinosulfon, dinoterbon,
DNOC, amitraz, chlordimeform,
chloromebuform, formetanate, formparanate, medimeform, semiamitraz,
afoxolaner, fluralaner, sarolaner, tetmnactin
oavennectin acaricides, abamectin, doramectin, eprinomectin, ivennectin,
selamectin, milbemectin, milbemycin oxime,
moxidectinõ clofentezine, cyromazine, diflovidazin, dofenapyn, fluazuron,
flubenzimine, flucycloxuron, flufenoxuron,
hcxythiazox, bromocycicn, camphcchlor, DDT, dicnochlor, cndosulfan, lindanc,
chlorfcnvinphos, crotoxyphos,
dichlorvos, heptenophos, mevinphos, monocrotophos, naled, TEPP,
tetrachlorvinphos, amidithion, amiton, azinphos-
ethyl, azinphos-methyl. azothoatc, bcnoxafos, bromophos, bromophos-ethyl,
carbophcnothion, chlorpyrifos,
cfflorthiophos, coumaphos, cyanthoate, demeton-O, demeton-S, demeton-O-methyl,
demeton-S-methyl, demeton-S-
methylsulphon, dialifos, diazinon, dimethoate, dioxathion, disulfoton,
endothion, ethion, ethoate-methyl, formothion,
malathion, mecarbam, methacrifos, omethoate, oxydeprofos, oxydisulfoton,
parathion, phenkapton, phorate, phosalone,
phosmet, phostin, phoxim, pirimiphos-methyl, prothidathion, prothoate,
pyrimitate, quinalphos, quintiofos, sophamide,
sulfotep, thiometon, triazophos, trifenofos, vamidothion. trichlorfon,
isocarbophos, methamidophos, propetamphos,
dimefox, mipafox, schradanõ azocyclotin, cyhexatin, fenbutatin oxide, phostinõ
dichlofluanid, dialifos, phosmet,
cyenopyrafen, fenpyroximate, pyflubumide, tebufenpyrad, acetoprole, fipronil,
vaniliproleõ acrinathrin, bifenthrin,
brofluthrinate, cyhalothrin, alpha-cypermethrin, fenpropathrin, fenvalerate,
flucythrinate, flumethrin, tau-fluvalinate,
permcthrin, halfcnprox, pyrimidifcn, chlorfenapyr, sanguinarinc,
chinomcthionat, thioquinox, bifujunzhi, fluacrypyrim,
flufenoxystrobin, pyriminostrobinõ aramite, propargite, spirodiclofen,
clofentezine, diflovidazin, flubenzimine,
hexythiazox, fenothiocarb, chloromethiuron, diafenthiuron, acequinocyl,
amidoflumet, arsenous oxide, clenpirin,
closantel, crotamiton, cycloprate, cymiazole, disulfiram, etoxazole,
fenazaflor, fenazaquin, fluenetil, mesulfen, MNAF,
nifluridide, nikkomycins, pyridaben, sulfiram, sulfluramid, sulfur,
thuringiensin, triarathene, and combinations thereof);
fungicides (e.g., strobilurins, such as azoxystrobin, coumethoxystrobin,
coumoxystrobin, dimo,x3Tstrobin, enestroburin,
fluoxastrobin, kresoxim-methyl, metominostrobin, orysastrobin, picoxystrobin,
pyraelostrobin, pyrametostrobin,
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pyraoxystrobin, pyribencath, trifloxystrobin, 242-(2,5-dimethyl-phenoxymethyl)-
pheny1J-3-methoxy-acrylic acid methyl
ester and 2-(2-(3-(2,6-dichloropheny1)-1-methyl-allylideneaminooxymethyl)-
pheny1)-2-methoxyimino-N-methyl-
acetamide; carboxamides, such as carboxanilides (e.g., benalaxyl, benalaxyl-M,
benodanil, bixafen, boscalid, carboxin,
fenfuram, fenhexamid, flutolanil, fluxapyroxad, furametpyr, isopyrazam,
isotianil, kiralaxyl, mepronil, metalaxyl,
metalaxyl-M (mefenoxam), ofurace, oxadixyl, oxycarboxin, penflufen,
penthiopyrad, sedaxane, tecloftalam,
thifluzamide. tiadinil, 2-amino-4-methyl-thiazole-5-carboxanilide. N-(4'-
trifluoromethylthiobipheny1-2-y1)-3-
difluommethyl-l-methyl-1H-pyra- zole-4-carboxamide, N-(2-(1,3,3-
trimethylbuty1)-pheny1)-1,3-dimethyl-5-fluoro-1H-
pyrazole-4-carboxamide), carboxylic morpholides (e.g., dimethomorph, flumorph,
pyrimorph), benzoic acid amides
(e.g., flumetover, fluopicolide, fluopyram, zoxamide), carpropamid,
dicyclomet, mandiproamid, oxytetracyclin,
silthiofam and N-(6-methoxy-pyridin-3-y1) cyclopropanecarboxylic acid amide;
azoles, such as triazoles (e.g.,
azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole,
diniconazole, diniconazole-M, epoxiconazole,
fenbuconazole, fluquinconazole, flusilazole, flutriafol, hexaconazole,
imibenconazole, ipconazole, metconazole,
myclobutanil, oxpoconazole, paclobutrazole, penconazole, propiconazole,
prothioconazole, simeconazole, tebuconazole,
tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole) and
imidazoles (e.g., cyazofamid, imazalil,
pefurazoate, prochloraz, triflumizol); heterocyclic compounds, such as
pyridines (e.g., fluazinam, pyrifenox (cf.D lb), 3-
[5-(4-chloro-pheny1)-2,3-dimethyl-isoxazolidin-3-y1]-pyridine, 345-(4-methyl-
pheny1)-2,3-climethyl-isoxazolidin-3-y1]-
pyridine), pyrimidines (e.g., bupirimate, cyprodinil, diflumetorim, fenarimol,
ferimzone, mepanipyrim, nitrapyrin,
nuarimol, pyrimethanil), piperazines (e.g., triforine), pirroles (e.g.,
fenpiclonil, fludioxonil), morpholines (e.g.,
aldimorph, dodemorph, dodemorph-acetate, fenpropimorph, tridemolph),
piperidines (e.g., fenpropidin), dicarboximides
(e.g., fluoroimid, iprodione, procymidone, vinclozolin), non-aromatic 5-
membered heterocycles (e.g., famoxadone,
fenamidone, flutianil, octhilinone, probenazole, 5-amino-2-isopropy1-3-oxo-4-
ortho-toly1-2,3-dihydro-pyrazole-1-
carbothioic acid S-allyl ester), acibenzolar-S-methyl, amctoctradin,
amisulbrom, anilazin, blasticidin-S, captafol, captan,
chinomethionat, dazomet, debacarb, diclomezine, difenzoquat, difenzoquat-
methylsulfate, fenoxanil, Folpet, oxolinic
acid, piperalin, proquinazid, pyroquilon, quinoxyfcn, triazoxidc,
tricyclazolc, 2-butoxy-6-iodo-3-propylchromen-4-one,
5-chloro-1-(4,6-dimethoxy-pyrimidin-2-y1)-2-methy1-1H-benzoimidazole and 5-
chloro-7-(4-methylpiperidin-l-y1)-6-
(2,4,6-trifluoropheny1)-[1,2,4]triazolo-[1,5-alpyrimidine; benzimidazoles,
such as carbendazim; and other active
substances, such as guanidines (e.g., guanidine, dodine, dodine free base,
guazatine, guazatine-acetate, iminoctadine),
iminoctadine-triacetate and iminoctadine-tris(albesilate); antibiotics (e.g.,
kasugamycin, kasugamycin hydrochloride-
hydrate, streptomycin, polyoxine and validamycin A); nitrophenyl derivates
(e.g., binapacryl, dicloran, dinobuton,
dinocap, nitrothal-isopropyl, tecnazen); organometal compounds (e.g., fentin
salts, such as fentin-acetate, lentil' chloride,
fentin hydroxide); sulfur-containing heterocyclyl compounds (e.g., dithianon,
isoprothiolane); organophosphorus
compounds (e.g., edifenphos, fosetyl, fosetyl-aluminum, iprobenfos, phosphorus
acid and its salts, pyrazophos, tolclofos-
methyl); organochlorinc compounds (e.g., chlorothalonil, dichlofluanid,
dichlorophcn, flusulfamidc, hexachlorobenzene,
pencycuron, pentacMorphenole and its salts, phthalide, quintozene, thiophanate-
methyl, thiophanate, tolylfluanid, N-(4-
chloro-2-nitro-pheny1)-N-ethy1-4-methyl-benzenesulfonamide), inorganic active
substances (e.g., Bordeaux mixture,
copper acetate, copper hydroxide, copper oxychloride, basic copper sulfate,
phosphite salt, sulfur, zinc sulfate),
natamycin, and combinations thereof); gastropodicides (e.g., methiocarb,
metaldehvde, carbaryl, spinosad, copper sulfate
in combination with lime, boric acid, iron phosphate, and combinations
thereof); herbicides (e.g., 2,4-
dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-
T), ametryn, amicarbazone,
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aminocyclopyrachlor, acetochlor, acifluorfen, alachlor, atrazine, azafenidin,
bentazon, benzofenap, bifenox, bromacil,
bromo.xynil, butachlor, butafenacil, butroxydim, carfentrazone-ethyl,
chlorimuron, chlorotoluro, clethodim, clodinafop,
clomazone, cyanazine, cycloxydim, cyhalofop, desmedipham, desmetryn, dicamba,
diclofop, dimefuron, diuron,
dithiopyr, fenoxaprop, fluazifop, fluazifop-P, fluometuron, flufenpyr-ethyl,
flumiclorac-pentyl, flumioxazin,
fluoroglycofen, fluthiacet- methyl, fomesafe, fomesafen, glyphosate,
glufosinate, haloxyfop, hexazinone, imazamox,
imazaquin, imazethapyr, ioxynil. isoproturon, isoxaflutole, lactofen, linuron,
mecoprop, mecoprop-P, mesotrion,
metamitron, metazochlor, methibenzuron , metolachlor (and S-metolachlor ),
metoxuron, metribuzin, monolinuron,
oxadiargyl, oxadiazon, oxyfluorfen, phenmedipham, pretilachlor, profoxydim,
prometon, prometry, propachlor,
propanil propaquizafop, propisochlor, pyraflufen-ethyl, pyrazon, pyrazolynate,
pyrazovfen, pyridate, quizalofop,
quizalofop-P (e.g., quizalofop-ethyl, quizalofop-P-ethyl, clodinafop-proparul,
cyhalofop-butyl, diclofop- methyl,
fenoxaprop-P-ethyl, fluazifop-P-butyl, haloxyfop-methyl, haloxyfop-R-methyl),
saflufenacil, sethoxydim, siduron,
simazine, simetryn, sulcotrione, sulfentrazone, tebuthiuron, tembotrione,
tepraloxydim, terbacil, terbumeton,
terbuthylazine, thaxtomin (e.g., the thaxtomins described in US Patent No.:
7,989,393), thenylchlor, tralkoxydim,
triclopyr, trietazine, tropramezone, salts and esters thereof; racemic
mixtures and resolved isomers thereof and
combinations thereof); and insecticides and nematicidcs (e.g., antibiotic
insecticides such as allosamidin and
thuringiensin; macrocyclic lactone insecticides such as spinosad, spinetoram,
and other spinosyns including the 21-
butenyl spinosyns and their derivatives; avermectin insecticides such as
abamectin, doramectin, emamectin,
eprinomectin, ivermectin and selamectin; milbemycin insecticides such as
lepimectin, milbemectin, milbemycin oxime
and moxidectin; arsenical insecticides such as calcium arsenate, copper
acetoarsenite, copper arsenate, lead arsenate,
potassium arsenite and sodium arsenite; other biological insecticides, plant
incorporated protectant insecticides such as
Cry lAb, Cry lAc, Cry1F, Cry1A.105, Cry2Ab2, Cry3A, mir Cry3A, Cry3Bb1, Cry34,
Cty35, and VIP3A; botanical
insecticides such as anabasinc, azadirachtin, d-limonene, nicotine,
pyrethrins, cincrins, cincrin I, cincrin II, jasmolin
jasmolin II, pyrethrin I, pyrethrin II, quassia, rotenone, iyania and
sabadilla; carbamate insecticides such as bendiocarb
and carbaryl; benzofuranyl methylcarbamate insecticides such as bcnfuracarb,
carbofuran, carbosulfan, decarbofuran and
furathiocarb; dimethylcarbamate insecticides dimitan, dimetilan, hyquincarb
and pirimicarb; oxime carbamate
insecticides such as alanycarb, aldicarb, aldoxycarb, butocarboxim,
butoxycarboxim, methomyl, nitrilacarb, oxamyl,
tazimcarb, thiocarboxime, thiodicarb and thiofanox; phenyl methylcarbamate
insecticides such as allyxycarb, aminocarb,
bufencarb, butacarb, carbanolate, cloethocarb, dicresyl, dioxacarb. EMPC,
ethiofencarb, fenethacarb, fenobucarb,
isoprocarb, methiocalb, metolcarb, mexacarbate, promacyl, promecatb, propoxur,
trimethacarb, )(MC and xylylcarb;
dinitrophenol insecticides such as dinex, dinoprop, dinosam and DNOC; fluorine
insecticides such as barium
hexafluorosilicate, cryolite, sodium fluoride, sodium hexafluorosilicate and
sulfluramid; formamidine insecticides such
as amitraz, chlordimeform, formetanate and formparanate; fumigant insecticides
such as acrylonitrile, carbon disulfide,
carbon tetrachloride, chloroform, chloropicrin, para-dichlorobenzene, 1,2-
dichloropropanc, ethyl formate, ethylene
dibromide, ethylene dichloride, ethylene oxide, hydrogen cyanide, iodomethane,
methyl bromide, methylchloroform,
methylene chloride, naphthalene, phosphine, sulfuryl fluoride and
tetrachloroethane; inorganic insecticides such as
borax, calcium polysulfide, copper oleate, mercurous chloride, potassium
thiocyanate and sodium thiocyanate; chitin
synthesis inhibitors such as bistrifluoron, buprofezin, chlorfluazuron,
cyromazine, diflubenzuron, flucycloxuron,
flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, penfluoron,
teflubenzuron and triflumuron; juvenile
hormone mimics such as epofenonane, fenoxycarb, hydroprene, kinoprene,
methoprene, pyriproxyfen and triprene;
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juvenile hormones such as juvenile hormone 1, juvenile hormone 11 and juvenile
hormone 111; moulting hormone agonists
such as chromafenozide, halofenozide, metho,xyfenozide and tebufenozide;
moulting hormones such as .alpha.-ecdysone
and ecdysterone; moulting inhibitors such as diofenolan; precocenes such as
precocene 1, precocene 11 and precocene Ill;
unclassified insect growth regulators such as dicyclanil; nereistoxin analogue
insecticides such as bensultap, cartap,
thiocyclam and thiosultap; nicotinoid insecticides such as flonicamid;
nitroguanidine insecticides such as clothianidin,
dinotefuran, imidacloprid and thiamethoxam; nitromethylene insecticides such
as nitenpyram and nithiazine;
pyridylmethylamine insecticides such as acetamiprid, imidacloprid, nitenpyram
and thiacloprid; organochlorine
insecticides such as bromo-DDT, camphechlor, DDT, pp'-DDT, ethyl-DDD, HCH,
gamma-HCH, lindane,
methoxychlor, pentachlorophenol and TDE; cyclodiene insecticides such as
aldrin, bromocyclen, chlorbicyclen,
chlordane, chlordecone, dieldrin, dilor, endosulfan, endrin, HEOD, heptachlor,
HHDN, isobenzan, isodrin, kelevan and
mirex; organophosphate insecticides such as bromfenvinfos, chlorfenvinphos,
croto,xyphos, dichlorvos, dicrotophos,
dimethylvinphos, fospirate, heptenophos, methocrotophos, mevinphos,
monocrotophos, naled, naftalofos,
phosphamidon, promphos, 'TEPP and tetrachlorvinphos; organothiophosphate
insecticides such as dioxabenzofos,
fosmethilan and phenthoate; aliphatic organothiophosphate insecticides such as
acethion, amiton, cadusafos,
chlorcthoxyfos, chlormcphos. dcmephion. demephion-0, demephion-S, demeton,
demeton-0, demcton-S, dcmeton-
methyl, demeton-O-methyl, demeton-S-methyl, demeton-S-methylsulphon,
disulfoton, ethion, ethoprophos, IPSP,
isothioate, malathion, methacrifos, oxydemeton-methyl, oxydeprofos,
oxydisulfoton, phorate, sulfotep, terbufos and
thiometon; aliphatic amide organothiophosphate insecticides such as
amidithion, cyanthoate, dimethoate, ethoate-methyl,
formothion, mecarbam, omethoate, prothoate, sophamide and vamidothion; oxime
organothiophosphate insecticides such
as chlorphoxim, phoxim and phoxim-methyl; heterocyclic organothiophosphate
insecticides such as azamethiphos,
coumaphos, coumithoate, dioxathion, endothion, menazon, morphothion,
phosalone, pyraclofos, pyridaphenthion and
quinothion; bcnzothiopyran organothiophosphatc insecticides such as
dithicrofos and thicrofos; benzotriazine
organothiophosphate insecticides such as azinphos-ethyl and azinphos-methyl;
isoindole organothiophosphate
insecticides such as dialifos and phosmet; isoxazolc organothiophosphate
insecticides such as isoxathion and zolaprofos;
pyrazolopyrimidine organothiophosphate insecticides such as clilorprazophos
and pyrazophos; pyridine
organothiophosphate insecticides such as chlorpyrifos and chlorpyrifos-methyl;
pyrimidine organothiophosphate
insecticides such as butathiofos, diazinon, etrimfos, lirimfos, pirimiphos-
ethyl, pirimiphos-methyl, primidophos,
pyrimitate and tebupirimfos; quinoxaline organothiophosphate insecticides such
as quinalphos and quinalphos-methyl;
thiadiazole organothiophosphate insecticides such as athidathion,
lythidathion, methidathion and prothidathion; triazole
organothiophosphate insecticides such as isazofos and triazophos; phenyl
organothiophosphate insecticides such as
azothoate, bromophos, bromophos-ethyl, carbophenothion, chlorthiophos,
cyanophos, cythioate, dicapthon,
dichlofenthion, etaphos, famphur, fenchlorphos, fenitrothion fensulfothion,
fenthion, fenthion-ethyl, heterophos,
jodfcnphos, mesulfenfos, parathion, parathion-methyl, phenkapton, phosnichlor,
profenofos, prothiofos, sulprofos,
temephos, trichlormetaphos-3 and trifenofos; phosphonate insecticides such as
butonate and ttichlotfon;
phosphonothioate insecticides such as mecarphon; phenyl ethylphosphonothioate
insecticides such as fonofos and
trichloronat; phenyl phenylphosphonothioate insecticides such as cyanofenphos,
EPN and leptophos; phosphoramidate
insecticides such as crufomate, fenamiphos, fosthietan, imicyafos,
mephosfolan, phosfolan and pirimetaphos;
phosphoramidothioate insecticides such as acephate, isocarbophos, isofenphos,
methamidophos and propetamphos;
phosphorodiamide insecticides such as dimefox, mazidox, mipafox and schradan;
oxadiazine insecticides such as
88
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indoxacalb; phthalimide insecticides such as dialifos, phosmet and
tetramethrin; pyrazole insecticides such as acetoprole,
ethiprole, fipronil, pyrafluprole, pyriprole, tebufenpyrad, tolfenpyrad and
vaniliprole; pyrethroid ester insecticides such
as acrinathrin, allethrin, bioallethrin, barthrin, bifenthrin,
bioethanomethrin, cyclethrin, cycloprothrin, cyfluthrin, beta-
cyfluthrin, cyhalothrin, gamma-cyhalothrin, lambda-cyhalothrin, cypermethrin,
alpha-cypermethrin, beta-cypermethrin,
theta-cypermethrin, zeta-cypermethrin, cyphenothrin, deltamethrin,
dimefluthrin, dimethrin, empenthrin, fenfluthrin,
fenpirithrin, fenpropathrin, fenvalemte, esfenvalemte, flucythrinate,
fluvalinate, tau-fluvalinate, furethrin, imiprothrin,
metofluthrin, pennethrin, biopennethrin, transpennethrin, phenothrin,
prallethrin, proflutluin, pyresmethrin, resmethrin,
biopermethrin, cismethrin, tefluthrin, terallethrin, tetramethrin,
tralomethrin and transfluthrin; pyrethroid ether
insecticides such as etofenprox, flufenprox, halfenprox, protrifenbute and
silafluofen; pyrimidinamine insecticides such
as flufenerim and pyrimidifen; pyrrole insecticides such as chlorfenapyr;
tetronic acid insecticides such as spirodiclofen,
spiromesifen and spirotetramat; thiourea insecticides such as diafenthiuron;
urea insecticides such as flucofuron and
sulcofuron; and unclassified insecticides such as AKD-3088,
chlorantraniliprole, closantel, crotamiton, cyflumetofen,
E2Y45, EXD, fenazaflor, fenazaquin, fenoxacrim, fenpyroximate, FKT-1033,
flubendiamide, HGW86, hydramethylnon,
IKI-2002, isoprothiolane, malonoben, metaflumizone, metoxadiazone,
nifluridide, NNI-9850, NNI-0101, pymetrozine,
pyridabcn, pyridalyl, pyrifluquinazon, Qcidc, rafoxanidc, Rynaxypyr.TM., SYJ-
159, triarathcnc and triazamatc, and
combinations thereof). It is to be understood that fommlation of the present
disclosure may comprise any suitable
combination of chemical actives and may therefore comprise two, three, four,
five, six, seven, eight, nine, ten or more of
the aforementioned actives. Conversely, in some embodiments, one, two, three,
four, five, six, seven, eight, nine, ten or
more of the aforementioned actives are expressly excluded from formulation of
the present disclosure.
Non-limiting examples of chemical active compositions that may be incorporated
into formulations of the
present disclosure¨or into which proteins and other compositions of the
present disclosure may be incorporated¨
include, but arc not limited to, commercial products sold under the tradenames
AGCELENCE , REVYSOLO,
XEMIUMIDz , INITIUM , F 500 , CLEARFIELD , KIXOR and SERIFEL from BASF
(Ludwigshafen, Germany);
CORVUSO, POWERMAX , DELAROO, PROSAROO, BAYTHROIDO, SIVANTO , FINISH ,
GINSTARO,
ACCELERON , RAXIL , AERIS , EVERGOL , TRILEX , ALLEGIANCE , BUTEO, EMESTO ,
GAUCHO ,
PONCHO and THIRAM from Bayer Crop Science (Creve Coeur, MO, USA); ABUNDIT ,
ACCENT ,
AFFORIA , APROACH , BASIS , BEXFOND , BLACKHAWK , CANOPY , CINCH , CLINHERO,
CURTAIL , CURZATE , DELEGATE . RAINSHIELD , DITHANE , FEXAPAN , VAPORGRIP ,
LANNATEO, TANOSO, DURANGO , DMA , ELEVORE , EMBED , ENABLE , ENLIST DUO ,
ENLIST
ONE , ENLITEV, ENTRUST , ENV1VE , EVERPLEX , FONTEL1S , FULT1ME , GOLD SKY V,
GRAND STAND , GRANITE , GRASP , HEARKEN , INDAR NXT GEN 41, INSTINCT ,
INTREPID 2E4-.3),
INTREPID EDGE , KERB , KEYSTONE , KYBER , LEADOFF , LOYANT , MATRIX , N-SERVE
,
NOVIXIDO, OPENSKY , PERFECTMATCHO, PINDARO, PIXXARO , POWERFLEXO, QUELEXO,
RADIANT , RALLY , REALM , REBELEX , RESICORE , RESOLVE , REVULIN , REZUVANT ,
RIDGEBACK , SEQUOIA , SIMPLICITY , SONIC , STARANE , SlEADFAST , STINGER ,
STRONGARMO, SUCCESS , SURESTART , SURPASS , SURVEIL , SYNCHRONY , TARZEC ,
TRANSFORM , TRELLIS , TRIVENCE , UTRISHA , VERTISAN , VYDATE , WIDEARMATCH ,
WIDEMATCH and ZEST from Corteva Agroscience (Indianapolis, IN, USA); BIO-
SAVE from Decco U.S. Post-
Harvest, Inc. (Monrovia, CA, USA); ALTACORliz, ATHENA , AVAUNT , BELEAF ,
BRIGADE , CARBINE ,
89
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CORAGEN , ELEVEST , EX1REL , GLADIATOR , HERO , MUSTANG , PREVATHON , STEWARD
,
VANTACORCD, VERIMARKCD, LUCENTO , RHYME , ROVRAL , TOPGUARDCD, TERRA , XYWAY ,
AFFINITY , AGILITY , AIM , ALLY , ANTHEM , AUTHORITY , CAPTURE , ETHOS ,
TEM1TRY ,
CADET , COMMAND , DISPLAY , EXPRESS , FINESSE , FIRSTSHOT , HARMONY , MARVEL ,
OBEY , PANOFLEX , SHARK , SOLIDA , SPARTAN , UPBEET , and ZEUS , from FMC
Corporation
(Philadelphia, PA, USA); PENTIACD, ABAMEXCD, AGRI TIN , CHAMP , CHIPTOX , GIN
OUT , KAISOCD,
MEPEX , NUPRID , RAPPORT , 1LRMINATE , THISTROL , ULTRA FLOURISH , GOAL ,
GOALTENDER , GRAPPLE , TUSCANY , CHAMPION++, AGRI-MYCIN , PHOSTROL , BLIGHTBAN
,
CHEETAH , MYCOSHIELD , RITEWAY , TAZER , MYSTIC , CUPROXAT and TYPY from
Nufarm
Limited (Victoria, Australia); B1OSPECTRA , PACR1TE , EFOG , SHIELD-BRITE ,
FUNGAFLOR ,
PENBO1LC, and SOPP from Pace International (Wapato, WA, USA); ALUMNI ,
CHAIRMAN , GRADUA
GRADUAIEA+ , MENTOR , 1VIERTECT , SCHOLAR and STADIUM from Syngenta Crop
Protection (Basel,
Switzerland); and ASULOXV3), BALTSTIKV3), BEETUP , BELLMAGIt, BETASANA ,
BETTTX , BUGUIS ,
CENTURION , CLIOPHARt, COLZAMID , CORZAL , DEFIANT , DEVRINOL , MINSTREL ,
AFFIX ,
AXIDORCD, BUZZ , MIMIXCD, DIOZINOSO, DIPROSPEROCD, EVITOO, MANZATE ,
MICROTHIOLCD,
NAUTILE , PENNCOZEB , PROMESS , PROPLANT , PROXANIL , PYRUS R, SACRON , SYLLIT
,
FEBUZOL , THIOPRON , TOKYO , UNIZEB CO), VACCIPLANT , VIDEO , ZOXIS ,
CYTHRINEW,
DIMILIN , FORESTER , FUMICYP , TALISMA , B-NINE , FAZORt, GYRO , HIMALAYA ,
ICENI ,
TRINEXIS , IODUS , AUDIT , BASAGRAN , BATLIUM , BOYCOTT , BROADLOOM , COYOTE ,
COLLIDE , DUET , ETHOTRONCD, EVEREST , IMIFLEX , LIFELINE , METRICORCD,
MOCCASIN ,
MOTIF , PRE-PARE , SATELLITE , SHADOW , SHUTDOWN , STAM , SUPERWHAM!
SUPREMACY , TRICORCD, TRIZENTACD, CUPROFIXCD, DEXTER , ELEVATE , ELIXIR ,
FORTIX ,
FROGHORN , ME1EOR , MICROTHI , ORANIL , PH-D , PROCURE , RANCOVA , TEPERA ,
TERRAGUARD R, TERRAMASTERO, TERRAZOLE R, TOPSINO, TRIONIC , ZIRAMCD, ZOLERA ,
ADIOS ,
GOLDWING , OFF-SHOOT-T , PACZOL , ROYAL , ROYALTAC , ACENTHRIN , ACEPHATE ,
ACRAMITE , ADEPT , ARGYLE , AS BANTER , BIFENTURE , BIOMI
__________________ IL R, COMI [ER, DIMILIN ',
ENKOUNTER , INTRUDER , KANEMITE , LAMBDA-CY , MICROMITE , ()MITE , PEDESTAL ,
PERM-
UP R, RIMON , STRAFER , TURNSTYLE , UP-CYDE , VENDEX , VIGILANT , ZYLO ,
ATTENDANT ,
BEAN GUARD , ALLEGIANCE , BELMONT , ENHANCE , GRAINGUARDO, MESH , PRO-GRO ,
RANCONA , STARTUP , TH1RAM , VITAFLO , V1TAVAX , MAGNAPHOS , WEEVIL-CIDER,,
AQUASTRIKE , AQUATHOL , PEGASUS, GOLIATH, POACONSTRICTOR, RAVEN, T-BIRD, UP-
END , UP-
START , ETHEPHON PEGASUS, GOLIATH, POACONSTRICTOR, RAVEN, T-BIRD, UP-END , UP-
START R,
ZEBA and FLORAMITEO from UPL Limited (Mumbai, Maharashtra, India).
Examples of dispersants (spreaders) that may be included in formulations of
the present disclosure include, but
not are not limited to, anionic surfactants, cationic surfactants and non-
ionic surfactants. See generally, e.g., EP 0245970;
US 5496568; WO 2008/144024; WO 2009/135049; WO 2011/126832; WO 2017/083049; WO
2020/225276; WO
2021/055316; WO 2022/096688; WO 2022/096691; WO 2022/096692; WO 2022/096693;
WO 2022/096694;
WO/2022/096695; WO 2022/096696.
In some embodiments, formulations of the present disclosure comprise one or
more anionic surfactants. For
CA 03223679 2023- 12- 20
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PCT/US2022/073761
example, in some embodiments, formulations of the present disclosure comprise
one or more anionic surfactants selected
from the group consisting of alkyl carboxylates (e.g., sodium stearate), alkyl
sulfates (e.g., alkyl lauryl sulfate, sodium
lauryl sulfate), alkyl ether sulfates, alkyl amido ether sulfates, alkyl aryl
polyether sulfates, alkyl aryl sulfates, alkyl aryl
sulfonates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates,
alkyl benzene sulfonates, alkyl diphenyloxide
sulfonate, alpha-olefin sulfonates, alkyl naphthalene sulfonates, paraffin
sulfonates, alkyl sulfosuccinates, alkyl ether
sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfosuccinamates, alkyl
sulfoacetates, alkyl phosphates, alkyl ether
phosphates, acyl sarconsinates, acyl isethionates, N-acyl taurates, N-acyl-N-
alkyltaurates, benzene sulfonates, cumene
sulfonates, dioctyl sodium sulfosuccinate, ethoxylated sulfosuccinates, lignin
sulfonates, linear alkylbenzene sulfonates,
monoglyceride sulfates, perfluorobutanesulfonate, perfluorooctanesulfonate,
phosphate ester, styrene acrylic polymers,
toluene sulfonates and xylene sulfonates.
In some embodiments, formulations of the present disclosure comprise one or
more cationic surfactants. For
example, in some embodiments, formulations of the present disclosure comprise
one or more cationic surfactants
selected from the group consisting of alkyltrimethylammonium salts (e.g.,
cetyl trimethylammonium bromide, cetyl
trimethylammonium chloride), cetylpyridinium chloride, benzalkonium chloride,
benzethonium chloride, 5-Bromo-5-
nitro-1,3-dioxanc, dimethyldioctadecylammonium chloride, cctrimonium bromide,
dioctadecyldimethylammonium
bromide and/or octenicline dihydrochloride.
In some embodiments, formulations of the present disclosure comprise one or
more nonionic surfactants. For
example, in some embodiments, formulations of the present disclosure comprise
one or more nonionic surfactants
selected from the group consisting of alcohol ethovlates (e.g.. TERGITOLTm 15-
S surfactants (The Dow Chemical
Company, Midland, MI), such as TERGITOLTm15-S-9, alkanolamides, alkanolamine
condensates, carboxylic acid
esters, cetostearyl alcohol, cetyl alcohol, cocamide DEA, dodecyldimethylamine
oxides, ethanolamides, ethoxylates of
glycerol ester and glycol esters, ethylene oxide polymers, ethylene oxide-
propylene oxide copolymers, glucosidc alkyl
ethers, glycerol alkyl ethers, glycerol esters, glycol alkyl ethers (e.g.,
polyoxyethylene glycol alkyl ethers,
polyoxypropylenc glycol alkyl ethers), glycol alkylphenol ethers (e.g.,
polyoxyethylene glycol alkylphenol ethers),
glycol esters, monolaurin, pentaethylene glycol monododecyl ethers, poloxamer,
polyamines, polyglycerol
polyricinoleate, polysorbate, polyoxyethylenated fatty acids,
polyoxyethylenated mercaptans, polyoxyethylenated
polyoxyproylene glycols, polyoxyethylene glycol sorbitan alkyl esters,
polyethylene glycol-polypropylene glycol
copolymers, polyoxyethylene glycol octylphenol ethers, polyvinyl pynolidones,
sugar-based alkyl polyglycosides,
sulfoanylamides, sorbitan fatty acid alcohol ethovlates, sothitan fatty acid
ester ethoxylates, sorbitan fatty acid ester
and/or tertiary acetylenic glycols.
In some embodiments, formulations of the present disclosure comprise one or
more zwitterionic surfactants. For
example, in some embodiments, formulations of the present disclosure comprise
one or more zwitterionic surfactants
selected from the group consisting of 34(3-cholamidopropyl)dinicthylammonio1-1-
propanesulfonate, cocamidopropyl
betaine, cocamidopropyl hydroxysultaine, phosphatidylserine,
phosphatidylethanolamine, phosphatidylcholine and/or
one or more sphingomyelins.
In some embodiments, formulations of the present disclosure comprise one or
more soaps and/or organosilicone
surfactants.
Non-limiting examples of dispersants that may be incorporated into
formulations of the present disclosure¨or
into which proteins and other compositions of the present disclosure may be
incorporated¨include ATLOXTm (e.g.,
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4916, 4991; Croda International PLC, Edison, NJ), ATLOX METASPERSErm (Croda
International PLC, Edison, NJ),
BIO-SOFT (e.g., N series, such as N1-3, N1-7, N1-5, N1-9, N23-3, N2.3-6.5,
N25-3, N25-7, N25-9, N91-2.5, N91-6,
N91-8; Stepan Company, Northfield, IL), MA_KON nonionic surfactants (e.g., DA-
4, DA-6 and DA-9; Stepan
Company, Northfield, IL), MORWET powders (Akzo Nobel Surface Chemistry LLC,
Chicago, IL), MULTIWETTm
surfactants (e.g., MO-85P-PW-(AP); Croda International PLC, Edison, NJ),
SILWETI-z) L-77 (Helena Chemical
Company, Collierville, TN), SPANTM surfactants (e.g., 20, 40, 60, 65, 80 and
85; Croda Inc., Edison NJ), TA1VIOLTm
dispersants (The Dow Chemical Company, Midland, MI), IERGITOLTm surfactants
(e.g., TMN-6 and TMN-100X; The
Dow Chemical Company, Midland, MI), TERSPERSE surfactants (e.g., 2001, 2020,
2100, 2105, 2158, 2700, 4894 and
4896; Hunstman Corp., The Woodlands, TX), TRITONTm surfactants (e.g., X-100;
The Dow Chemical Company,
Midland, M1), TWEEN surfactants (e.g., TWEEN 20, 21, 22, 23, 28, 40, 60, 61,
65, 80, 81 and 85; Croda
International PLC, Edison, NJ) and combinations thereof. Additional examples
of dispersants may be found in BAIRD &
ZUBLENA. 1993. Sou_ FACTS: USING WETTING AGENTS (NONIONIC SURFACTANTS) ON SOIL
(North Carolina Cooperative
Extension Service Publication AG-439-25) (1993); BURGES, FORMULATION OF
MICROBIAL BTOPESTECIDES: BENEFICIAL
MICROORGANISMS, NEMATODES AND SEED TREATMENTS (Springer Science & Business
Media) (2012); MCCARTY,
WETTING AGENTS (Clemson University Cooperative Extension Service Publication)
(2001).
Examples of drying agents that may be included in formulations of the present
disclosure include, but not are
not limited to, drying powders. Non-limiting examples of drying agents include
AEROSIL hydrophobic fumed silica
powders (Evonik Corporation, Parsippany, NJ), BENTOLITE powders (BYK-Chemie
GmbH, Wesel, Germany),
INCOTEC powders (INCOTEC Inc., Salinas, CA), SIPERNAT silica powders (Evonik
Corporation, Parsippany, NJ)
and combinations thereof. Additional examples of drying agents may be found in
BURGES, FORMULATION OF MICROBIAL
BIOPESTICIDES: BENEFICIAL MICROORGANISMS, NEMATODES AND SEED TREATMENTS
(Springer Science & Business
Media) (2012). In some embodiments, compositions of the present disclosure
comprise calcium stearate, clay (e.g.,
attapulgite clay, montmorillonite clay), graphite, magnesium stearate,
magnesium sulfate, powdered milk, silica (e.g.,
fumed silica, hydrophobically-coated silica, precipitated silica), soy
lecithin and/or talc.
Examples of microbes that may be included in formulations of the present
disclosure include, but not are not
limited to, diazotrophs, phosphate-solubilizing microorganisms and
biopesticides. See generally, e.g., WO 92/08355; US
2003/082164; US 2008/320615; US 2016/345588; US 2018/168168; US 2005/187107;
US 2006/258534; US
2018/279624; US 10820594; US 2019/014786; US 10874109; US 10856552; US
2019/014787; US 11076603; US
2020/093125; US 2020/085065; US 2020/000098; US 2019/345572; US 2020/263734;
WO 2021/101949; WO
2021/101937; WO 2016/201284; WO 2018/186307; WO 2007/142543; WO 2017/205800;
WO 2015/003908; WO
2021/018321; WO 2003/016510; WO 2016/044542; WO 92/11856.
In some embodiments, formulations of the present disclosure comprise one or
more of the following:
Azospirillum brasilense INTA Az-39, Bacillus amyloliquefaciens D747, Bacillus
amyloliquefaciens NRRL B 50349,
Bacillus amyloliquefaciens TJ1000, Bacillus amyloliquefaciens FZB24, Bacillus
amyloliquefaciens FZB42, Bacillus
amyloliquefaciens IN937a, Bacillus ainyloliquefaciens IT-45, Bacillus
amyloliquefaciens TJ1000, Bacillus
amyloliquefaciens MBI600, Bacillus amyloliquefaciens BS27 (deposited as NRRL B-
5015), Bacillus amyloliquefaciens
BS2084 (deposited as NRRL B-50013), Bacillus amyloliquefaciens 15AP4
(deposited as ATCC PTA-6507), Bacillus
amyloliquefaciens 3AP4 (deposited as ATCC PTA-6506), Bacillus
amyloliquefaciens LSSA01 (deposited as NRRL B-
50104), Bacillus amyloliquefaciens ABP278 (deposited as NRRL B-50634),
Bacillus amyloliquefaciens 1013 (deposited
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as NRRL B-50509), Bacillus amyloliquefaciens 918 (deposited as NRRL B-50508),
Bacillus amyloliquefaciens 22CP1
(deposited as ATCC PTA-6508) and Bacillus amyloliquelaciens B S18 (deposited
as NRRL B-50633), Bacillus cereus I-
1562, Bacillus firmu,s 1-1582, Bacillus lichenjOrmi,s BA842 (deposited as NRRL
B-50516), Bacillus lichenjOrmis BL21
(deposited as NRRL B-50134), Bacillus mycoides NRRL B-21664, Bacillus pumilus
NRRL B 21662, Bacillus plaudits
NRRL B-30087, Bacillus pumilus ATCC 55608, Bacillus pumilus ATCC 55609,
Bacillus pumilus GB34, Bacillus
pumilus KFP9F. Bacillus pumilus QST 2808, Bacillus subtilis ATCC 55078,
Bacillus subtilis ATCC 55079, Bacillus
subtilis MBI 600, Bacillus subtilis NRRL B-21661, Bacillus subtilis NRRL B-
21665, Bacillus subtilis CX-9060,
Bacillus subtilis GB03, Bacillus subtilis GB07, Bacillus subtilis QST-713,
Bacillus subtilis FZB24, Bacillus subtilis
D747, Bacillus subtilis 3BP5 (deposited as NRRL B-50510), Bacillus
thuringiensis ATCC 13367, Bacillus thuringiensis
GC-91, Bacillus thuringiensis NRRL B-21619, Bacillus thuringiensis ABTS-1857,
Bacillus thuringiensis SAN 4011,
Bacillus thuringiensis ABG-6305, Bacillus thuringiensis ABG-6346. Bacillus
thuringiensis AI\465-52, Bacillus
thuringiensis SA-12, Bacillus thuringiensis SB4, Bacillus thuringiensis ABTS-
351, Bacillus thuringiensis HD-1,
Bacillus thuringiensis EG 2348, Bacillus thuringiensis EG 7826, Bacillus
thuringiensis EG 7841, Bacillus thuringiensis
DSM 2803, Bacillus thuringiensis NB-125, Bacillus thuringiensis NB-176,
Bradyrhizobium spp. 8A57, Bradyrhizobium
elkanii SEMIA 501, Bradyrhizobium elkanii SEMIA 587, Bradyrhizobium elkanii
SEMIA 5019, Bradyrhizobium
japonicum 61A227, Bradyrhizobium japonicum 61A228, Bradyrhizobium japonicum
61A273, Bradyrhizobium
japonicum E-109, Bradyrhizobium japonicum NRRL B-50586 (also deposited as NRRL
B-59565), Bradyrhizobium
japonicum NRRL B-50587 (also deposited as NRRL B-59566), Bradyrhizobium
japonicum NRRL B-50588 (also
deposited as NRRL B-59567), Bradyrhizobium japonicum NRRL B-50589 (also
deposited as NRRL B-59568),
Bradyrhizobium japonicum NRRL B-50590 (also deposited as NRRL B-59569),
Bradyrhizobium japonicum NRRL B-
50591 (also deposited as NRRL B-59570), Bradyrhizobium japonicum NRRL B-50592
(also deposited as NRRL B-
59571), Bradyrhizobium japonicum NRRL B-50593 (also deposited as NRRL B-
59572), Bradyrhizobium japonicum
NRRL B-50594 (also deposited as NRRL B-50493), Bradyrhizobium japonicum NRRL B-
50608, Bradyrhizobium
japonicum NRRL B-50609, Bradyrhizobium japonicum NRRL B-50610, Bradyrhizobium
japonicum NRRL B-50611,
Bradyrhizobium japonicum NRRL B-50612, Bradyrhizobium japonicum NRRL B-50726,
Bradyrhizobium japonicum
NRRL B-50727, Bradyrhizobium japonicum NRRL B-50728, Bradyrhizobim n japonicum
NRRL B-50729,
Bradyrhizobium japonicum NRRL B-50730, Bradyrhizobium japonicum SEMIA 566,
Bradyrhizobium japonicum
SEMIA 5079, Bradyrhizobium japonicum SEMIA 5080, Bradyrhizobium japonicum USDA
6, Bradyrhizobium
japonicum USDA 110, Bradyrhizobium japonicum USDA 122, Bradyrhizobium
japonicum USDA 123, Bradyrhizobium
japonicum USDA 127, Bradyrhizobium japonicum USDA 129, Bradyrhizobium
japonicum USDA 532C, Gliocladium
virens ATCC 52045, Gliocladium virens GL-21, Glomus intraradices RTI-801,
Metarhizium anisopliae F52,
Penicillium bilaiae ATCC 18309, Penicillium bilaiae ATCC 20851, Penicillium
bilaiae ATCC 22348, Penicillium
bilaiae NRRL 50162, Penicillium bilaiae NRRL 50169, Penicillium bilaiae NRRL
50776, Penicillium bilaiae NRRL
50777, Penicillium bilaiae NRRL 50778, Penicillium bilaiae NRRL 50777,
Penicillium bilaiae NRRL 50778,
Penicillium bilaiae NRRL 50779, Penicillium bilaiae NRRL 50780, Penicillium
bilaiae NRRL 50781, Penicillium
bilaiae NRRL 50782, Penicillium bilaiae NRRL 50783, Penicillium bilaiae NRRL
50784, Penicillium bilaiae NRRL
50785, Penicillium bilaiae NRRL 50786, Penicillium bilaiae NRRL 50787,
Penicillium bilaiae NRRL 50788,
Penicillium bilaiae RS7B-SD1, Penicillium brevicompactum AgRF18, Penicillium
canescens ATCC 10419, Penicillium
expansum ATCC 24692, Penicillium expansum YT02, Penicillium fellatanum ATCC
48694, Penicillium gaestrivorns
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NRRL 50170 , Penicillium glabrum DAOM 239074, Penicillium glabrum CBS 229.28,
Penicillium janthinellum ATCC
10455, Penicillium lanosocoeruleum ATCC 48919, Penicillium radicum ATCC
201836, Penicillium radicum FRR
4717, Penicillium radicum FRR 4719, Penicillium radicum N93/47267, Penicillium
raistrickil ATCC 10490,
Pseudomonas jessenii PS06, Rhizobium leguminosarum SO12A-2 (IDAC 080305-01),
Sinorhizobium fredii
CCBAU114, Sinorhizobium fredii USDA 205, Trichoderma asperellum SKT-1,
Trichoderma asperellum ICC 012,
Trichoderma atroviride LC52, Trichoderma atroviride CNCM 1-1237, Trichoderma
fertile JM41R, Trichoderma gamsii
ICC 080, Trichoderma hamatum ATCC 52198, Trichoderma harzianum ATCC 52445,
Trichoderma harzianum KRL-
AG2, Trichoderma harzianum T-22, Trichoderma harzianum TH-35, Trichoderma
harzianum T-39, Trichoderma
harzianum ICC012, Trichoderma reesi ATCC 28217, Trichoderma virens ATCC 58678,
Trichoderma virens G1-3,
Trichoderma virens GL-21. Trichoderma virens G-41, Trichoderma viridae ATCC
52440, Trichoderma viridae ICC080,
Trichoderma viridae TV1, Yersinia entomophaga strain 043NEW (NRRL B-67598),
Yersinia entomophaga strain
024G3R (NRRL B-67599), Yersinia enlomophaga strain 024KEK (NRRL B-67600) and
Yersinia enlomophaga strain
0333A4 (NRRL B-67601).
Non-limiting examples of microbial compositions that may be incorporated into
formulations of the present
disclosure-or into which proteins and other compositions of the present
disclosure may be incorporated-include, but
are not limited to, commercial products sold under the tradenames SERIFEL
from BASF (Ludwigshafen, Germany);
VITIVO from Bayer Crop Science (Creve Coeur, MO, USA); BIONIQ , CELL-TECH ,
JUIVWSTART ,
NITRAGIN , OPTIMIZE , QUICKROOTS and TAGTEAM from Novozymes North America,
Inc. (Durham, NC,
USA).
Examples of nutrients that may be included in formulations of the present
disclosure include, but not are not
limited to, organic acids (e.g., acetic acid, citric acid, lactic acid, malic
acid, taurine, etc.), macrominerals (e.g.,
phosphorous, calcium, magnesium, potassium, sodium, iron, etc.), trace
minerals (e.g., boron, cobalt, chloride,
chromium, copper, fluoride, iodine, manganese, molybdenum, selenium, zinc,
etc.), vitamins, (e.g., vitamin A, vitamin B
complex (i.e., vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6,
vitamin B7, vitamin B8, vitamin B9, vitamin
B12, choline) vitamin C, vitamin D, vitamin E, vitamin K, carotenoids (a-
carotene, 0-carotene, ciyptoxanthin, lutein,
lycopene, zeaxanthin, etc.), and combinations thereof. See also, generally,
e.g., US 2014/235447; WO 2022/029224; WO
2022/029221; WO 2021/255118; WO 2021/247915; US 2021/300837; US 2020/148605;
US 2012/247164; US
2016/355443; US 2020/055794; US 2011/154873; US 2006/243009; US 2017/088474.
Examples of pest attractants and feeding stimulants that may be included in
formulations of the present
disclosure include, but not are not limited to, brevicomin, ceralure,
codlelure, cue-lure, disparlure, dominicalure,
eugenol, frontalin, gossyplure, grandlure, hexalure, ipsdienol, ipsenol,
japonilure, latitlure, lineatin, litlure, looplure,
medlure, megatomic acid, methyl eugenol, moguchun, a-multistriatin, muscalure,
orfalure, oryctalure, ostramone,
rescalure, siglurc, sulcatol, trimcdlure, trunc-call, and combinations
thereof. See generally, e.g., WO 00/28824; US
5607684; US 4510133; US 5290556; US 60774634; US 6773727; WO 2022/051661; EP
0563963; WO 2013/164384;
WO 2003/020030; US 8420070; US 5401506; WO 92/11856.
Examples of postharvest treatments that may be included in formulations of the
present disclosure include, but
not are not limited to, essential oils, ethylene biosynthesis inhibitors
(e.g., cyclopropenes), pesticides and waxes. See
generally, e.g., WO 00/10386; WO 01/43548; US 2002/058592; US 2002/061822; US
2002/043730; US 2002/198107;
US 2004/072694; US 2003/100450; US 2004/077502; US 2005/043179; US
2005/250649; US 2005/261131; US
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2005/261132; US 2005/288189; CA 2512254; CA 2512256; US 2007/117720; US
2007/265166; US 2008/113867; US
2010/144533; US 2008/206823; US 2009/035380; U52009/077684; US 2009/118492; US
2009/230350; US
2010/047408; US 2011/321191; US 2011/034335; US 2011/014334; US 2012/272572;
US 2012/282380;
US2011/293801; US2012/004108; US2013/065764; US2012/258220; US2012/142534; US
2014/127309; US
2013/004634; US 2014/011679; US 2015/208679; WO 2014/120715; WO 2015/175157;
WO 2017/180695; US
2014/080710; US 2015/237877; US 2015/272115; US 2016/000072; US 2015/366189;
US 2014/242235; US
2014/271758; US 2016/066568; US 2016/095311; US 2015/018430; US 2015/087520;
US 2015/231588; US
2015/366230; US 2016/235070; US 2016/324147; US 2017/251673; US 2017/251662;
US 2017/251669; US
2017/265462; US 2017/318804; US 2018/139975; US 2018/356384; US 2021/102245;
US 2021/238201; WO
2022/094214; EP 2468107; ES 2439616; WO 2014/128321; WO 2018/116027; WO
2018/128807; WO 2019/058211;
WO 2020/157714; US 2009/253578; US 2009/253579; US 2004/146617; US
2022/087261; WO 2020/016728; US
2010/173773; US 2010/292080; US 5858436; EP 0972450; US 6221414; US 6723364;
WO 00/49880; US 6403139; US
2005/129662; US 2006/228458; US 2005/137090; US 2006/276336; US 2008/175926;
US 2008/016766; US
2008/145499; US 2010/092631; US 2010/081636; US 2011/003694; US 2011/008475;
US 2010/298147; US
2013/072383; US 2013/156835; US 2013/266670; US 2013/236562; US 2013/178489;
US 2014/187570; US
2013/306158; US 2013/341809; US 2016/330987; WO 2017/001502; US 2019/159469;
WO 2017/220581; US
2020/060300; US 2020/221719; WO 2020/016154; WO 2020/225066; WO 2021/233900;
WO 2005/058014.
Non-limiting examples of postharvest treatment compositions that may be
incorporated into formulations of the
present disclosure¨or into which proteins and other compositions of the
present disclosure may be incorporated¨
include commercial products sold under the tradenames ACTISEALTm, ETHYLBLOC,
FRESHSTART,
SMARTCITRUS, TEYCER, VITAFRESH from AgroFresh, Inc. (Philadelphia, PA, USA);
CERAXEL ,
CERASULFURO, CERAQUINTO, ELIMO, MUSACARE and CERAFRUTA from CERADIS Crop
Protection
(Ceradis B.V., Netherlands); APL-LUSTR , APL-BRITE, BIO-SAVE , CITRUBLUSH,
CITRUS BRITE, CITRUS
FIXTM, CITRUS LUSTRII, DECCO and DECCONATURTm from Decco U.S. Post-Harvest,
Inc. (Monrovia, CA, USA);
SEMPERFRESH, LUSTRE DRY, NATURAL SHINE , PACRITE , PRIMAFRESH , SHIELD-BRITE ,
EFOG
and XEDAQUIN from Pace International (Wapato, WA, USA); ALUMNI , CHAIRMAN ,
GRADUATE ,
GRADUATEA+t, MENTOR , MERTECT , SCHOLAR and STADIUM from Syngenta Crop
Protection (Basel,
Switzerland); and MAGNAPHOS, QUICKPHLO-R, QUICKPHOS from UPL Limited (Mumbai,
Maharashtra, India).
It is to be understood that many of the compounds described above as "chemical
actives" and "chemical active
compositions" may be applied to plants and plant parts both preharvest and
postharvest and may therefore also be
considered "postharvest treatments" and "postharvest treatment compositions."
Examples of suitable UV protectants include, but not are not limited to,
aromatic amino acids (e.g., tryptophan,
tyrosinc), carotcnoids, cinnamatcs, lignosulfonatcs (e.g., calcium
lignosulfonatc, sodium lignosulfonatc), melanins,
mycosporines, polyphenols and/or salicylates). Non-limiting examples of UV
protectants include Bonegaard
LignoTechTm lignosulfonates (e.g., Borresperse 3A, Borresperse CA, Borresperse
NA, Marasperse AG, Norlig A, Norlig
11D, Ufoxane 3A, Ultrazine NA, Vanisperse CB; Borregaard Lignotech, Sarpsborg,
Norway) and combinations thereof.
Additional examples of UV protectants may be found in BURGES, FORMULATION OF
MICROBIAL BIOPESTICIDES:
BENEFICIAL MICROORGANISMS, NEMATODES AND SEED TREATMENTS (Springer Science &
Business Media) (2012).
Examples of suitable wetting agents include, but are not limited to,
naphthalene sulfonates, such as alkyl
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naphthalene sulfonates (e.g., sodium alkyl naphthalene sulfonate), isopropyl
naphthalene sulfonates (e.g., sodium
isopropyl naphthalene sulfonate) and butyl naphthalene sulfonates (e.g.,
sodium n-butyl naphthalene sulfonate).
It is to be understood that formulations of the present disclosure may
comprise combinations of the enzymes,
including, but not limited to combinations of enzymes expressly disclosed
herein and combinations with enzymes that are
not expressly disclosed herein.
In some embodiments, formulations of the present disclosure comprise 2, 3, 4,
5, 6, 7, 8, 9, 10 or more proteins
of the present disclosure. For example, in some embodiments, formulations of
the present disclosure comprise 2, 3, 4, 5,
6, 7, 8, 9, 10 or more of the polypeptides set forth herein as SEQ ID Nos: 1-
48 and 98-150 or a mature polypeptide thereof.
It is to be understood that formulations of the present disclosure may
comprise one or more filler and/or carrier
materials to increase the volume and improve the handleability of the
formulations. Suitable filler or carrier materials for
granule/particle formulations include, but are not limited to, various salts
of carbonate, chloride, silicate and sulfate, as
well as talc, clay and the like. Suitable filler or carrier materials for
liquid formulation include, but are not limited to, water
or low molecular weight primary and secondary alcohols including polyols and
diols. Examples of such alcohols include,
but are not limited to, methanol, ethanol, propanol and isopropanol. In some
embodiments, formulations of the present
disclosure comprise about 5% to about 90% w/w of such filler/carrier
materials.
In some preferred embodiments, formulations of the present disclosure,
comprise, consist essentially of or consist
of:
a) about 0.0001 to about 25 %w/w protein (e.g., one or more proteins of the
present disclosure), more preferably
about 0.0001 to about 5 `)/0 w/w, even more preferably about 0.001 to about 1
`)/0 w/w;
b) about 25 to about 75 % w/w enzyme stabilizer (e.g., one or more polyols,
such as glycerol and/or sorbitol),
more preferably about 25 to about 50 % vv/w, even more preferably about 35 to
about 45 % w/w;
c) about 0.001 to about 1 % w/w preservative (e.g., potassium sorbatc
and/or sodium benzoate), more preferably
about 0.01 to about 1 % w/w, even more preferably 0.1 to about 0.5 %; and
d) water.
In some preferred embodiments, formulations of the present disclosure,
comprise, consist essentially of or consist
of:
a) about 0.0001 to about 25 %w/w protein (e.g., one or more proteins of the
present disclosure), more preferably
about 0.0001 to about 5 `)/0 w/w, even more preferably about 0.001 to about 1
`)/0 w/w;
b) about 0.001 to about 1 % w/w preservative (e.g., potassium sorbate
and/or sodium benzoate), more preferably
about 0.01 to about 1 % w/w, even more preferably 0.1 to about 0.5 %; and
c) water.
In some preferred embodiments, formulations of the present disclosure,
comprise, consist essentially of or consist
of:
a) about 0.0001 to about 25 %w/w protein (e.g., one Or more proteins of the
present disclosure), more preferably
about 0.0001 to about 5 % w/w, even more preferably about 0.001 to about 1 %
w/w;
b) about 25 to about 75 % w/w enzyme stabilizer (e.g., one or more polyols,
such as glycerol and/or sorbitol),
more preferably about 25 to about 50 % vv/w, even more preferably about 35 to
about 45 % w/w; and
c) water.
In some preferred embodiments, fommlations of the present disclosure,
comprise, consist essentially of or consist
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of:
a) about 0.0001 to about 5 % w/w protein (e.g., one or more proteins of the
present disclosure), more preferably
about 0.0001 to about 1 % w/w, even more preferably about 0.0001 to about 0.5
% w/w;
b) about 0.001 to about 10 % w/w rain fastener (e.g., one or more organo-
modified siloxanes, such as organo-
modified trisiloxanes and organo-modified polysiloxanes), more preferably
about 0.025 to about 1 % w/w, even more
preferably about 0.025 to about 0.5 % w/w;
c) about 0.001 to about 5 % w/w pH control component (e.g., one or more
acetate, carbonate, citrate and/or
phosphate buffers), more preferably about 0.01 to about 1 % w/w, even more
preferably 0.05 to about 0.5 %; and
d) water.
In some preferred embodiments, formulations of the present disclosure,
comprise, consist essentially of or consist
of:
a) about 0.0001 to about 5 % w/w protein (e.g., one or more proteins of the
present disclosure), more preferably
about 0.0001 to about 1 % w/w, even more preferably about 0.0001 to about 0.5
% w/w;
b) about 0.001 to about 10 % w/w rain fastener (e.g., one or more organo-
modified siloxanes, such as organo-
modified trisiloxanes and organo-modified polysiloxanes), more preferably
about 0.025 to about 1 % w/w, even more
preferably about 0.025 to about 0.5 A) w/w;
c) about 0.001 to about 5 % w/w pH control component (e.g., one or more
acetate, carbonate, citrate and/or
phosphate buffers), more preferably about 0.01 to about 1 % w/w, even more
preferably 0.05 to about 0.5 %;
d) about 1 to about 5 `)/0 w/w enzyme stabilizer (e.g., one or more polyols,
such as glycerol and/or sorbitol),
more preferably about 1 to about 2.5 % w/w, even more preferably about 1 to
about 2 % w/w; and
e) water.
As demonstrated in the Examples set forth below, combinations of enzymes may
be particularly useful for
preventing, treating, suppressing, eliminating and/or reducing the severity of
infestations/infections of/by horticultural
pests, such as acarids, bacteria, fungi, gastropods, insects, nematodes,
oomycetes, protozoa and weeds.
In some embodiments, formulations of the present disclosure comprise two or
more distinct celluloses.
In some embodiments, formulations of the present disclosure comprise two or
more distinct glucanases.
In some embodiments, formulations of the present disclosure comprise two or
more distinct hemicellulases.
In some embodiments, formulations of the present disclosure comprise two or
more distinct pectinases.
In some embodiments, formulations of the present disclosure comprise two or
more distinct peptidases.
In some embodiments, formulations of the present disclosure comprise two or
more distinct proteases.
In some embodiments, formulations of the present disclosure comprise two or
more distinct xylanases.
In some embodiments, formulations of the present disclosure comprise at least
one cellulose and at least one
hemicellulase.
In some embodiments, formulations of the present disclosure comprise at least
one amylase, at least one glucanase,
and at least one bacillolycin.
In some embodiments, formulations of the present disclosure comprise at least
one cellulose, at least one
hemicellulase, and at least one xylanase.
In some embodiments, formulations of the present disclosure comprise at least
one cellulose, at least one
glucanase, and at least one pectinase.
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In some embodiments, formulations of the present disclosure comprise at least
one cellulase, at least one
glucanase, and at least one xylanase.
In some embodiments, formulations of the present disclosure comprise at least
one cellulase and at least one
xylanase.
In some embodiments, formulations of the present disclosure comprise at least
one furanosidase and at least one
xylanase.
In some embodiments, formulations of the present disclosure comprise at least
one hemicellulase and at least one
xylanase.
In some embodiments, formulations of the present disclosure comprise at least
one peptidase and at least one
protease.
In some embodiments, formulations of the present disclosure comprise a
fermentation broth that comprises 2, 3,
4, 5, 6, 7, 8, 9, 10 or more enzymes.
in some embodiments, formulations of the present disclosure comprise a
fermentation broth that comprises 2, 3,
4, 5, 6, 7, 8, 9, 10 or more proteins of the present disclosure.
The present disclosure also provides polynucleotides encoding proteins of the
present disclosure, including, but
not limited to, nucleic acid constructs, recombinant expression vectors and
recombinant host cells that encode one or
more enzymes of the present disclosure, as well as methods of producing such
polynucleotides.
The polynucleotide may be a genomic DNA, a cDNA, a synthetic DNA, a synthetic
RNA, a mRNA, or a
combination thereof.
The polynucleotide may be cloned from any suitable genus, species or strain.
In some embodiments, the protein is cloned from a Grain-negative bacteria,
such as Campylobacter,
Dicytoglomus (e.g., D. thermophilum), Escherichia (e.g., E. coli),
Flavobacterium, Fuso bacterium, Helicobacter,
Ilyobacter, Neisseria, Pseudomonas, Salmonella or Ureaplasma.
In some embodiments, the polynucleotide is cloned from a Gram-positive
bacteria, such as Bacillus (e.g., B.
agaradhaerens, Bacillus alk-alophilus, Bacillus amyloliqzzefaciens, Bacillus
brevis, Bacillus circzzlans, Bacillus clausii,
Bacillus coagulans, B. deramificans, Bacillus firmus, Bacillus lawns, Bacillus
lentils, Bacillus licheniformis, Bacillus
megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis,
Bacillus thuringiensis), Clostridium,
Entero coccus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus (e.g.,
0. barbara), Staphylococcus,
Sfreptococcus (e.g., Streptococcus equisimilis, Streptococcus pyogenes,
Streptococcus uberis, and Streptococcus equi
subsp. Zooepidemicu,$) or Streptomyces (e.g., Streptomyces achromogene,s,
Streptomyces avermitill,s, Streptomyces
coelicolor, Streptomyces griseus, Streptomyces lividans).
In some embodiments, the polynucleotide is cloned from a fungus, such as
Acremonium, Aspergillus (e.g., A.
aculeatus, Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus,
Aspergillus japonicus, Aspergillus nidulans,
Aspergillus niger, Aspergillus oryzae), Aureolxisidium, Bjerkandem (e.g.,
Bjerhindera adusta), Ceriporiopsis (e.g.,
Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens,
Ceriporiopsis pannocinta, Ceriporiopsis
rivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora), Chaetomium
(e.g., C. erraticum), Chrysosporium (e.g.,
Chrysosporium mops, Chrysosporium keratinophilum, Chrysosporium lucknowense,
Chrysosporium merdarium,
Chrysosporium pannicokt, Chrysosporium queenslandicum, Chrysosporium tropicum,
Chrysosporium zonatum),
Coprinu,s (e.g., Coprirms cinereits), Coriolms (e.g., Coriolits hirsittu,$),
Cryptococcu,s, 1 FitSarilA111 (e.g.,
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Fusarium bactridioides, Fusarium cereal/s. Fusarium crookwellense, Fusarium
culmorum, Fusarium graminearum,
Fusarium graminum, Fusarium heterosporum, Fusarium negundi , Fusarium
oxysporum, Fusarium reticulatum, Fusarium
ro,seum, Fusarium ,sambucinum, Fusarium ,sarcochroum, F ,solani, Fusarium
,sporotrichioide,s, Fusarium ,sulphureum,
Fusarium torulosum, Fusarium trichothecioides, Fusarium venenatum), Hum/cola
(e.g., Hum/cola insolens, Hum/cola
lanuginosa), Alagnaporthe, Aficrodochium (e.g., Al. nivale), Alucor (e.g.,
Alucor miehei), Alyceliophthora (e.g.,
Mycehophthora thermophila), Neocallitnastix, Neurospora (e.g., Neurospora
crassa), Paecilomyces, Pen/c//hum (e.g.,
Pen/c/ purpurogenum), Phanerochaete (e.g., Phanerochaete
chrysosporium), Phlebia (e.g., Phlebia radiata),
Piromyces, Pleurotus (e.g., Pleurotus eryngii), Schizophyllum , Talaromyces
(e.g., Talaromyces emersonii), Thermoascus
(e.g., T. aurantiacus), Thielavia (e.g., Thielavia terrestris), Tolypocladium,
Trametes (e.g., Trametes villosa, Trametes
versicolor), or Trichoderma (e.g., T atroviride, Trichoderma harzianum,
Trichoderma koningii, Trichoderma
longibrachiatum, Trichoderma reesei, Trichoderma viride).
In some embodiments, the polynucleotide is cloned from a yeast, such as Candi
da, Hansen-Ida, Kluyveromyces
(e.g., Khtyveromyces lactic), PichiaõS'accharomyces (e.g.õS'accharomyces
carlsbergensisõS'accharomyces cerevisiae,
S'accharomyces diastaticus, S'accharomyces douglasii, Saccharomyces kluyveri,
S'accharomyces norbensis,
Saccharomyces oviforrnis), Schizosactharomyces, or Yarrowia (e.g., Yarrowia
hpolytica).
In some embodiments, the polynucleotide encodes:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any
one or more of SEQ ID NOs: 1-48 and 98-150;
b) a polypeptide having aboul/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to a
mature polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150;
c) a polypeptide derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
e) a polypeptide derived from a mature polypeptide any one
of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or insertion of one or more amino acids;
a polypeptide derived from the polypeptide of any one of a) through e) wherein
the N- and/or C-
terminal end has been extended by the addition of one or more amino acids;
and/or
a fragment of the polypeptide of any one of a) through f).
In preferred embodiments, the polynucleotide encodes a polypeptide that
comprises, consists essentially of or
consists of the amino acid sequence of any one of SEQ ID NOs: 1-48 and 98-150
or a mature polypeptide thereof.
In some embodiments, the polynucleotide comprises, consists essentially of or
consists of a nucleic acid sequence
having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % sequence identity
to any one or more of SEQ ID NOs: 49-97
and 151-203 or the cDNA sequence thereof;
In preferred embodiments, the polynucleotide comprises, consists essentially
of or consists of the nucleic acid
sequence of any one of SEQ ID NOs: 49-97 and 151-203 or a cDNA sequence
thereof.
In some embodiments, the polynucleotide encodes a polypeptide that comprises,
consists essentially of or
consists of a fragment of one of SEQ ID NOs: 1-48 and 98-150 or a mature
peptide thereof. For example, the
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polynucleotide may encode a fragment of any one of SEQ ID NOs: 1-48 and 98-150
or a mature peptide thereof, said
fragment comprising at least 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98 or 99 % of the amino acids found in the
original protein.
In some embodiments, polynucleotides of the present disclosure encode a
polypeptide that comprises, consists
essentially of or consists of any one of SEQ ID NOs: 1-48 and 98-150 or a
mature polypeptide thereof with an N-
terminal extension of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
more amino acids), a C-terminal extension of one or more amino acids (e.g., 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, 20 or more amino acids), one or more substitutions (e.g., 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19,20 or more substitutions), one or more insertions (e.g., 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20 or more insertions), and/or one or more deletions (e.g., 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15. 16, 17,
18, 19, 20 or more deletions). The amino acid changes may be of a minor
nature, that is conservative amino acid
substitutions or insertions that do not significantly affect the folding
and/or activity of the protein; small deletions,
typically of 1-30 amino acids; small amino- or carboxyl-terminal extensions,
such as an amino-terminal methionine
residue; a small linker peptide of up to 20-25 residues; or a small extension
that facilitates purification by changing net
charge or another function, such as a poly-histidinc tract, an antigenic
epitope or a binding module.
Polynucleotides of the present disclosure may be imitated by introduction of
nucleotide substitutions that do not
result in a change in the amino acid sequence of the protein, but which
correspond to the codon usage of the host organism
intended for production of the enzyme, or by introduction of nucleotide
substitutions that may give rise to a different amino
acid sequence. For a general description of nucleotide substitution, see,
e.g., Ford et al., 1991, Protein Expression and
Purification 2: 95-107.
In an aspect, the polynucleotide is isolated.
In another aspect, the polynucleotide is purified.
The present disclosure also relates to nucleic acid constructs comprising a
polynucleotide of the present disclosure,
wherein the polynucleotide is operably linked to one or more control sequences
that direct the expression of the coding
sequence in a suitable host cell under conditions compatible with the control
sequences.
The polynucleotide may be manipulated in a variety of ways to provide for
expression of the protein. Manipulation
of the polynucleotide prior to its insertion into a vector may be desirable or
necessary depending on the expression vector.
Techniques for modifying polynucleotides utilizing recombinant DNA methods are
well known in the art.
The control sequence may be a promoter, a polynucleotide that is recognized by
a host cell for expression of a
polynucleotide encoding a protein of the present disclosure. The promoter
contains transcriptional control sequences that
mediate the expression of the protein. The promoter may be any polynucleotide
that shows transcriptional activity in the
host cell including mutant, truncated, and hybrid promoters, and may be
obtained from genes encoding extracellular or
intracellular proteins either homologous or heterologous to the host cell.
Examples of suitable promoters for directing transcription of the
polynucleotide of the present disclosure in a
bacterial host cell are described in Sambrook et al., 1989, Molecular Cloning:
A Laboratory Manual, Cold Spring Harbor
Lab., NY, Davis et al., 2012, supra, and Song et al., 2016, PLOS One 11(7):
00158447.
Examples of suitable promoters for directing transcription of the
polynucleotide of the present disclosure in a
filamentous fungal host cell are promoters obtained from Aspergillus,
Fusarium, Rhizomucor and Trichoderma cells, such
as the promoters described in Mukherjee el al., 2013, "Trichoderma: Biology
and Applications", and by Schmoll and
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Dattenbock, 2016, "Gene Expression Systems in Fungi: Advancements and
Applications", Fungal Biology.
For expression in a yeast host, examples of useful promoters are described by
Smolke et al., 2018, "Synthetic
Biology: Parts, Devices and Applications" (Chapter 6: Constitutive and
Regulated Promoters in Yeast: How to Design and
Make Use of Promoters in S. cerevisiae), and by Schmoll and DattenbOck, 2016,
"Gene Expression Systems in Fungi:
Advancements and Applications", Fungal Biology.
The control sequence may also be a transcription terminator, which is
recognized by a host cell to terminate
transcription. The terminator is operably linked to the 3'-terminus of the
polynucleotide encoding the protein. Any
terminator that is functional in the host cell may be used in the present
disclosure.
Preferred terminators for bacterial host cells may be obtained from the genes
for Bacillus clausii alkaline protease
(aprH), Bacillus lichenifbrmis alpha-amylase (amyL), and Escherichia coli
ribosomal RNA (rrnB).
Preferred terminators for filamentous fungal host cells may be obtained from
Aspergillus or Trichoderma species,
such as obtained from the genes for Aspergillus niger glucoamylase,
Trichoderma reesei beta-glucosidase, Trichoderma
reesei cellobiohydrolase 1, and Trichoderma reesei endoglucanase T, such as
the terminators described in Mukherjee et al.,
2013, "Trichoderma: Biology and Applications", and by Schmoll and Dattenboek,
2016, "Gene Expression Systems in
Fungi: Advancements and Applications", Fungal Biology.
Preferred terminators for yeast host cells may be obtained from the genes for
Saccharomyces cerevisiae enolase,
Saccharomyces cerevisiae cytochrome C (CYC1), and Saccharomyces cerevisiae
glyceraldehyde-3-phosphate
dehydrogenase. Other useful terminators for yeast host cells are described by
Romanos et al., 1992, Yeast 8: 423-488.
The control sequence may also be an mRNA stabilizer region downstream of a
promoter and upstream of the
coding sequence of a gene which increases expression of the gene.
Examples of suitable mRNA stabilizer regions are obtained from a Bacillus
thuringiensis cryIHA gene (WO
94/25612) and a Bacillus subtihs 5P82 gene (Hue et al., 1995, J. Bacteriol.
177: 3465-3471).
Examples of mRNA stabilizer regions for fungal cells are described in Geisberg
et al., 2014, Cell 156(4): 812-
824, and in Morozov et al., 2006, Eukaryotic Cell 5(11): 1838-1846.
The control sequence may also be a leader, a non-translated region of an mRNA
that is important for translation
by the host cell. The leader is operably linked to the 5'-terminus of the
polynucleotide encoding the protein. Any leader
that is functional in the host cell may be used.
Suitable leaders for bacterial host cells are described by Hambraeus et al.,
2000, Microbiology 146(12): 3051-
3059, and by Kaberdin and Blasi, 2006, FEMS Microbiol. Rev. 30(6): 967-979.
Preferred leaders for filamentous fungal host cells may be obtained from the
genes for Aspergillus oryzae TAKA
amylase and Aspergillus nidulans triose phosphate isomerase.
Suitable leaders for yeast host cells may be obtained from the genes for
Saccharomyces cerevisiae enolase
(ENO-1), Saccharomyces cerevisiae 3 -pho sphoglycerate kinasc, Saccharomyces
cerevisiae alpha-factor, and
Saccharomyces cerevisiae alcohol dehydrogenase/glyceraldehyde-3-phosphate
dehydrogenase (ADH2/GAP).
The control sequence may also be a polyadenylation sequence, a sequence
operably linked to the 3' -terminus of
the polynucleotide which, when transcribed, is recognized by the host cell as
a signal to add polyadenosine residues to
transcribed mRNA. Any polyadenylation sequence that is functional in the host
cell may be used.
Preferred polyadenylation sequences for filamentous fungal host cells are
obtained from the genes for Aspergillus
nidulans anthranilate synthase, Aspergillus niger glucoamylase, Aspergillus
niger alpha-glucosidase, Aspergillus oryzae
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TAKA amylase, and Fusarium oxysporum trypsin-like protease.
Useful polyadenylation sequences for yeast host cells are described by Guo and
Sherman, 1995, Mol. Cellular
Biol. 15: 5983-5990.
The control sequence may also be a signal peptide coding region that encodes a
signal peptide linked to the
N-terminus of a protein and directs the protein into the cell's secretory
pathway. The 5'-end of the coding sequence of the
polynucleotide may inherently contain a signal peptide coding sequence
naturally linked in translation reading frame with
the segment of the coding sequence that encodes the protein. Alternatively,
the 5'-end of the coding sequence may contain
a signal peptide coding sequence that is heterologous to the coding sequence.
A heterologous signal peptide coding
sequence may be required where the coding sequence does not naturally contain
a signal peptide coding sequence.
Alternatively, a heterologous signal peptide coding sequence may simply
replace the natural signal peptide coding
sequence to enhance secretion of the protein. Any signal peptide coding
sequence that directs the expressed protein into
the secretory pathway of a host cell may be used.
Effective signal peptide coding sequences for bacterial host cells are the
signal peptide coding sequences obtained
from the genes for Bacillus NCIB 11837 maltogenic amylase. Bacillus
licheniformis subtilisin, Bacillus licheniformis beta-
lactamasc, Bacillus stearothermophilus alpha-amylase, Bacillus
stearothermophilus neutral protcascs (nprT nprS, nprM),
and Bacillus subtilis prsA. Further signal peptides are described by Freud!,
2018, Microbial Cell Factories 17: 52.
Effective signal peptide coding sequences for filamentous fungal host cells
are the signal peptide coding
sequences obtained from the genes for Aspergillus mger neutral amylase,
Aspergillus mger glucoamylase, Aspergillus
oryzae TAKA amylase, Hum/cola insolens cellulase, Humicola insolens
endoglucanase V. Humicola lanuginosa lipase,
and Rhizomucor miehei aspartic proteinase, such as the signal peptide
described by Xu el al., 2018, Biotechnology Letters
40: 949-955
Useful signal peptides for yeast host cells are obtained from the genes for
Saccharomyces cerevisiae alpha-factor
and Saccharomyces cerevisiae invertase. Other useful signal peptide coding
sequences are described by Romano s et al.,
1992, supra.
The control sequence may also be a propeptide coding sequence that encodes a
propeptide positioned at the
N-terminus of a protein. The resultant protein is known as a proenzyme or
proprotein (or a zymogen in some cases). A
proprotein is generally inactive and can be converted to an active protein by
catalytic or autocatalytic cleavage of the
propeptide from the proprotein. The propeptide coding sequence may be obtained
from the genes for Bacillus subtilis
alkaline protease (aprE), Bacillus subtilis neutral protease (nprT),
Myceliophthora thermophila laccase (WO 95/33836),
Rhizonntcor miehei aspartic proteinase, and Saccharomyce,s cerevisiae alpha-
factor.
Where both signal peptide and propeptide sequences are present, the propeptide
sequence is positioned next to
the N-terminus of a protein and the signal peptide sequence is positioned next
to the N-terminus of the propeptide sequence.
Additionally or alternatively, when both signal pcptidc and propcptidc
sequences arc present, the protein may comprise
only a part of the signal peptide sequence and/or only a part of the
propeptide sequence. Alternatively, the final or isolated
protein may comprise a mixture of mature proteins and proteins which comprise,
either partly or in full length, a propeptide
sequence and/or a signal peptide sequence.
It may also be desirable to add regulatory sequences that regulate expression
of the protein relative to the growth
of the host cell. Examples of regulatory sequences are those that cause
expression of the gene to be turned on or off in
response to a chemical or physical stimulus, including the presence of a
regulatory compound. Regulatory sequences in
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prokaryotic systems include the lac, tac, and trp operator systems. In yeast,
the ADH2 system or GAL1 system may be
used. In filamentous fungi, the Aspergillus niger glucoamylase promoter,
Aspergillus oryzae TAKA alpha-amylase
promoter, and A,spergillu,s oryzae glucoamylase promoter, Trichoderma reesei
cellobiohydrolase 1 promoter, and
Trichoderma reesei cellobiohydrolase II promoter may be used. Other examples
of regulatory sequences are those that
allow for gene amplification. In fungal systems, these regulatory sequences
include the dihydrofolate reductase gene that
is amplified in the presence of methotrexate, and the metallothionein genes
that are amplified with heavy metals.
The control sequence may also be a transcription factor, a polynucleotide
encoding a polynucleotide-specific
DNA-binding protein that controls the rate of the transcription of genetic
information from DNA to mRNA by binding to
a specific polynucleotide sequence. The transcription factor may function
alone and/or together with one or more other
proteins or transcription factors in a complex by promoting or blocking the
recruitment of RNA polymerase. Transcription
factors are characterized by comprising at least one DNA-binding domain which
often attaches to a specific DNA sequence
adjacent to the genetic elements which are regulated by the transcription
factor. The transcription factor may regulate the
expression of a protein of interest either directly, i.e., by activating the
transcription of the gene encoding the protein of
interest by binding to its promoter, or indirectly, i.e., by activating the
transcription of a further transcription factor which
regulates the transcription of the gene encoding the protein of interest, such
as by binding to the promoter of the further
transcription factor. Suitable transcription factors for fungal host cells are
described in WO 2017/144177. Suitable
transcription factors for prokaryotic host cells are described in Seshasayee
etal., 2011, Subcellular Biochemistry 52: 7-23,
as well in Balleza etal., 2009, FEMS Microbiol. Rev. 33(1): 133-151.
The present disclosure also relates to recombinant expression vectors
comprising a polynucleotide of the present
disclosure, a promoter, and transcriptional and translational stop signals.
The various nucleotide and control sequences
may be joined together to produce a recombinant expression vector that may
include one or more convenient restriction
sites to allow for insertion or substitution of the polynucleotide encoding
the protein at such sites. Alternatively, the
polynucleotide may be expressed by inserting the polynucleotide or a nucleic
acid construct comprising the polynucleotide
into an appropriate vector for expression. In creating the expression vector,
the coding sequence is located in the vector so
that the coding sequence is operably linked with the appropriate control
sequences for expression.
The recombinant expression vector may be any vector (e.g., a plasmid or virus)
that can be conveniently subjected
to recombinant DNA procedures and can bring about expression of the
polynucleotide. The choice of the vector will
typically depend on the compatibility of the vector with the host cell into
which the vector is to be introduced. The vector
may be a linear or closed circular plasmid.
The vector may be an autonomously replicating vector, i.e., a vector that
exists as an extrachromosomal entity,
the replication of which is independent of chromosomal replication, e.g., a
plasmid, an extrachromosomal element, a
minichromosome, or an artificial chromosome. The vector may contain any means
for assuring self-replication.
Alternatively, the vector may be one that, when introduced into the host cell,
is integrated into the gcnomc and replicated
together with the clu-omosome(s) into which it has been integrated.
Furthermore, a single vector or plasmid or two or more
vectors or plasmids that together contain the total DNA to be introduced into
the genome of the host cell, or a transposon,
may be used.
The vector preferably contains one or more selectable markers that permit easy
selection of transformed,
transfected, transduced, or the like cells. A selectable marker is a gene the
product of which provides for biocide or viral
resistance, resistance to heavy metals, prototrophy to auxotrophs, and the
like.
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The vector preferably contains at least one element that permits integration
of the vector into the host cell's
genome or autonomous replication of the vector in the cell independent of the
genome.
For integration into the host cell genome, the vector may rely on the poly
nucleotide's sequence encoding the
protein or any other element of the vector for integration into the genome by
homologous recombination, such as
homology-directed repair (HDR), or non-homologous recombination, such as non-
homologous end-joining (NHEJ).
For autonomous replicatioa the vector may further comprise an origin of
replication enabling the vector to
replicate autonomously in the host cell in question. The origin of replication
may be any plasmid replicator mediating
autonomous replication that functions in a cell. The term "origin of
replication- or "plasmid replicator- means a
polynucleotide that enables a plasmid or vector to replicate in vivo.
More than one copy of a polynucleotide of the present disclosure may be
inserted into a host cell to increase
production of a protein. For example, 2 or 3 or 4 or 5 or more copies are
inserted into a host cell. An increase in the copy
number of the polynucleotide can be obtained by integrating at least one
additional copy of the sequence into the host cell
genome or by including an amplifiable selectable marker gene with the
polynucleotide where cells containing amplified
copies of the selectable marker gene, and thereby additional copies of the
polynucleotide, can be selected for by cultivating
the cells in the presence of the appropriate selectable agent.
The present disclosure also provides cells expressing one or more proteins of
the present disclosure, including
microorganisms that naturally express one or more enzymes of the present
disclosure and microorganisms that have been
engineered to express one or more enzymes of the present disclosure.
In some embodiments, the cell expresses:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any
one or more of SEQ ID NOs: 1-48 and 98-150;
b) a polypeptide haying about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to a
mature polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 `)/0 sequence identity to any one or more of SEQ ID NOs: 49-97 and 151-203
or the cDNA sequence thereof;
d) a polypeptide derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
e) a polypeptide derived from a mature polypeptide any one of SEQ ID NOs: 1-
48 and 98-150 by
substitution, deletion or insertion of one or more amino acids;
f) a polypcptide derived from the polypeptide of any one of a) through c)
wherein the N- and/or C-
terminal end has been extended by the addition of one or more amino acids;
and/or
a fragment of the polypeptide of any one of a) through f).
In preferred embodiments, the cell expresses a polypeptide that comprises,
consists essentially of or consists of
the amino acid sequence of any one of SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof.
In some embodiments, the cell expresses a polypeptide that comprises, consists
essentially of or consists of a
fragment of one of SEQ ID NO: 1-48 and 98-150 or a mature peptide thereof. For
example, the microorganism may
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express a fragment of any one of SEQ ID NOs: 1-48 and 98-150 or a mature
peptide thereof, said fragment comprising
at least 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 % of the
amino acids found in the original protein.
In some embodiments, the cell expresses a polypeptide that comprises, consists
essentially of or consists of any
one of SEQ ID NOs: 1-48 and 98-150 or a mature polypeptide thereof with an N-
terminal extension of one or more
amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20 or more amino acids), a C-terminal
extension of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19,20 or more
amino acids), one or more substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
substitutions), one or more insertions (e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19,20 or more
insertions), and/or one or more deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
deletions). The amino acid changes may be of a minor nature, that is
conservative amino acid substitutions or insertions
that do not significantly affect the folding and/or activity of the protein;
small deletions, typically of 1-30 amino acids;
small amino- or carboxyl-temiinal extensions, such as an amino-terminal
methionine residue; a small linker peptide of
up to 20-25 residues; or a small extension that facilitates purification by
changing net charge or another function, such as
a poly-histidine tract, an antigenic epitope or a binding module.
In some embodiments, the cell comprises a homologous or heterologous nucleic
acid sequence that is at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the nucleic
acid sequences set forth herein as SEQ ID
Nos: 49-97 and 151-203 or the cDNA sequence thereof.
In preferred embodiments, the cell comprises a polynucleotide encodes a
polypeptide that comprises, consists
essentially of or consists of the amino acid sequence of any one of SEQ ID
NOs: 1-48 and 98-150 or a mature polypeptide
thereof.
In some embodiments, the cell comprises a polynucleotide that comprises,
consists essentially of or consists of a
nucleic acid sequence encoding a polypeptide having about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % sequence identity
to any one or more of SEQ ID NOs: 1-48 and 98-150.
In some embodiments, the cell comprises a polynucleotide that comprises,
consists essentially of or consists of a
nucleic acid sequence encoding a polypeptide having about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77,78. 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % sequence identity
to a mature polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150.
In preferred embodiments, the cell comprises a polynucleotide that comprises,
consists essentially of or consists
of the nucleic acid sequence of any one of SEQ ID NOs: 49-97 and 151-203 or a
cDNA sequence thereof.
In some embodiments, the cell comprises a polynucleotide that encodes a
polypeptide that comprises, consists
essentially of or consists of a fragment of one of SEQ ID NOs: 1-48 and 98-150
or a mature peptide thereof. For
example, the microorganism comprises a polynucleotide that encodes a fragment
of any one of SEQ ID NOs: 1-48 and
98-150 or a mature peptide thereof, said fragment comprising at least 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98 or 99 % of the amino acids found in the original protein.
In some embodiments, the cell comprises a polynucleotide that encodes a
polypeptide that comprises, consists
essentially of or consists of any one of SEQ ID NOs: 1-48 and 98-150 or a
mature polypeptide thereof with an N-
tenninal extension of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
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more amino acids), a C-terminal extension of one or more amino acids (e.g., 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, 20 or more amino acids), one or more substitutions (e.g., 1,
2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20 or more substitutions), one or more insertions (e.g., 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20 or more insertions), and/or one or more deletions (e.g., 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, 20 or more deletions). The amino acid changes may be of a minor
nature, that is conservative amino acid
substitutions or insertions that do not significantly affect the folding
and/or activity of the protein; small deletions,
typically of 1-30 amino acids; small amino- or carbox-yl-terminal extensions,
such as an amino-terminal methionine
residue; a small linker peptide of up to 20-25 residues; or a small extension
that facilitates purification by changing net
charge or another function, such as a poly-histidine tract, an antigenic
epitope or a binding module.
The present disclosure thus extends to recombinant host cells, comprising a
polynucleotide of the present
disclosure operably linked to one or more control sequences that direct the
production of a protein of the present disclosure.
A construct or vector comprising a polynucleotide is introduced into a host
cell so that the construct or vector is
maintained as a chromosomal integrant or as a self-replicating extra-
chromosomal vector as described earlier. The choice
of a host cell will to a large extent depend upon the gene encoding the
protein and its source. The protein can be native or
heterologous to the recombinant host cell. Also, at least one of the one or
more control sequences can be heterologous to
the polynucleotide encoding the protein. The recombinant host cell may
comprise a single copy, or at least two copies, e.g.,
three, four, five, or more copies of the polynucleotide of the present
disclosure.
The host cell may be any microbial cell useful in the recombinant production
of a protein of the present disclosure,
e.g., a prokaryotic cell or a fungal cell.
The prokaryotic host cell may be any Grain-positive or Grain-negative
bacterium. Grain-positive bacteria include,
but are not limited to, Bacillus, Clo,stridium, Enterococcu,s, Geobacillu,s,
Lactobacillus, Lactococcu,s, Oceanob acillu,s,
Staphylococcus, Streptococcus, and Streptomyces. Gram-negative bacteria
include, but are not limited to, Campylobacter,
E. coli, Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neisseria,
Pseudomonas, Sahnonella, and Ureaplasma.
The bacterial host cell may be any Bacillus cell including, but not limited
to, Bacillus alkalophilus, Bacillus
amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii,
Bacillus coagzilans, Bacillus firmus, Bacillus
lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus
pumilus, Bacillus stearothermophilus,
Bacillus subtilis, and Bacillus thuringiensis cells. In an embodiment, the
Bacillus cell is a Bacillus amylohquefaciens,
Bacillus lichendormis and Bacillus subtilis cell.
For purposes of this disclosure, Bacillus classes/genera/species shall be
defined as described in Patel and Gupta,
2020, Mt. J. ,Syst. Evol. Azficrobiol. 70: 406-438.
The bacterial host cell may also be any Streptococcus cell including, but not
limited to, Streptococcus equisimilis,
Streptococcus pyogenes, Streptococcus uberis, and Streptococcus equi subsp.
Zooepidemicus cells.
The bacterial host cell may also be any Streptomyces cell including, but not
limited to, Streptomyces
achromogenes, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces
griseus, and Streptomyces liviclans cells.
Methods for introducing DNA into prokaryotic host cells are well-known in the
art, and any suitable method can
be used including but not limited to protoplast transformation, competent cell
transformation, electroporation, conjugation,
transduction, with DNA introduced as linearized or as circular polynucleotide.
Persons skilled in the art will be readily
able to identify a suitable method for introducing DNA into a given
prokaryotic cell depending, e.g., on the genus. Methods
for introducing DNA into prokaryotic host cells are for example described in
Heinze et at, 2018, BAK:Microbiology 18:56,
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Burke et al., 2001, Proc. Natl. Acad. Sci. USA 98: 6289-6294, Choi et al.,
2006, J. Microbiol. Methods 64: 391-397, and
Donald etal., 2013, 1 Bacteriol. 195(11): 2612-2620.
The host cell may be a fungal cell. "Fungi" as used herein includes the phyla
Ascomycota, Basidiomycota,
Chytridiomycota, and Zygomycota as well as the Oomycota and all mitosporic
fungi (as defined by Hawksworth et al., In,
Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB
International, University Press, Cambridge, UK).
Fungal cells may be transformed by a process involving protoplast-mediated
transformation, Agrobacterium-
mediated transformation, electroporation, biolistic method and shock-wave-
mediated transformation as reviewed by Li et
al., 20 17 , Microbic-11 Cell Factories 16: 168 and procedures described in EP
238023, Yelton et al., 1984, Proc. Natl. Acad.
Sci. USA 81: 1470-1474, Christensen et al., 1988, Bio/Technology 6: 1419-1422,
and Lubertozzi and Keasling, 2009,
Biotechn. Advances 27: 53-75. However, any method known in the art for
introducing DNA into a fungal host cell can be
used, and the DNA can be introduced as linearized or as circular
polynucleotide.
The fungal host cell may be a yeast cell. "Yeast" as used herein includes
ascosporogenous yeast (Endomycetales),
basidiosporogenous yeast, and yeast belonging to the Fungi imperfecti
(Blastomycetes). For purposes of this disclosure,
yeast shall be defined as described in Biology and Activities of Yeast
(Skinner, Passmore, and Davenport, editors, Soc. App.
Bacteriol. Symposium Series No. 9, 1980).
The yeast host cell may be a Candida, Hansen ula, Kluyveromyces, Pichia,
Saccharomyces, S'chizosaccharomyces,
or Yarrowia cell, such as a Kluyveromyces lactis, Saccharomyces
carlsbergensis, Saccharomyces cerevisiae,
Saccharomyces chastaticus, Saccharomyces douglasit, Saccharomyces kluyvert,
Saccharomyces norbensts,
Saccharomyces ovijOrmis, or Yarrowia lipolytica cell. In a preferred
embodiment, the yeast host cell is a Pichia or
Komagataella cell, e.g., a Pichia pastoris cell (Komagalaella phajfii).
The fungal host cell may be a filamentous fungal cell. "Filamentous fungi"
include all filamentous forms of the
subdivision Eumycota and Oomycota (as defined by Hawksworth et al., 1995,
supra). The filamentous fungi are generally
characterized by a mycelial wall composed of chitin, cellulose, glucan,
chitosan, mannan, and other complex
polysaccharides. Vegetative growth is by hyphal elongation and carbon
catabolism is obligatcly aerobic. In contrast,
vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of
a unicellular tliallus and carbon catabolism
may be fermentative.
The filamentous fungal host cell may be an Acremonium, Aspergillus,
Aureobasidium, Bjerkandera,
Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium,
Fusarium, Humicola, Magnaporthe,
Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium,
Phanerochaete, Phlebia, Piromyces,
Pleurotu,s, Schizophyllum, 7alaromyce,s, Thermodscu,s, Thielavia,
Irolypocladium, Trametes, or Trichoderma cell. In a
preferred embodiment, the filamentous fungal host cell is an Aspergillus,
Trichoderma or Fusarium cell. In a further
preferred embodiment, the filamentous fungal host cell is anAspergillus niger,
Aspergillus oryzae,Trichoderma reesei, or
Fusarium venenatum cell.
For example, the filamentous fungal host cell may be an Aspergillus awamori,
Aspergillus foetidus, Aspergillus
fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger,
Aspergillus oryzae, Bjerkandera adusta,
Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens,
Ceriporiopsis pannocinta, Ceriporiopsis
rivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora, Chrysosporium
mops, Chrysosporium keratinophilum,
Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola,
Chrysosporium queenslandicum,
Chrysosporium tropicum, Chrysosporium zonatum, Coprinms cinereus, Coriolus
hirsunts, Fusarium bactridioides,
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Fusarium cereal/s. Fusarium crookwellense, Fusarium culmorum, Fusarium
graminearum, Fusarium graminum,
Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium
reticulatum, Fusarium roseum, Fusarium
sambucinum, Fusarium sarcochroum, Fusarium ,sporotrichioides, Fusarium
sulphureum, Eusarium torulosum, Fusarium
trichothecioides, Fusarium venenatum, Hum/cola insolens, Hum/cola lanuginosa,
Mucor miehei, Alyceliophthora
therm ophila, Neurospora crassa, purpurogenum, Phanerochaete
chtysosporium, Phlebia radiata, Pleurotus
eryngii, Talaromyces emersonii, Thielavia terrestris, Trametes villosa,
Trametes versicolor, Trichoderma harzianum,
Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, or
Trichoderma viride cell.
In an aspect, the host cell is isolated.
In another aspect, the host cell is purified.
The present disclosure also provides plants and plant parts expressing one or
more proteins of the present
disclosure, including plants that naturally express one or more enzymes of the
present disclosure and plants that have
been engineered to express one or more enzymes of the present disclosure.
in some embodiments, the plant or plant part expresses:
a) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to any
one or more of SEQ ID NOs: 1-48 and 98-150;
b) a polypeptide having about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % sequence identity to a
mature polypeptide of any one or more of SEQ ID NOs: 1-48 and 98-150;
c) a polypeptide encoded by a polynucleotide having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % sequence identity to any one or more of SEQ ID NOs: 49-97 and 151-203 or
the cDNA sequence thereof;
d) a polypeptide derived from any one of SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or
insertion of one or more amino acids;
e) a polypeptide derived from a mature polypeptide any one of SEQ ID NOs: 1-
48 and 98-150 by
substitution, deletion or insertion of one or more amino acids;
a polypeptide derived from the polypeptide of any one of a) through e) wherein
the N- and/or C-
terminal end has been extended by the addition of one or more amino acids;
and/or
a fragment of the polypeptide of any one of a) through f).
In preferred embodiments, the plant or plant part expresses a polypeptide that
comprises, consists essentially of
or consists of the amino acid sequence of any one of SEQ ID NOs: 1-48 and 98-
150 or a mature polypeptide thereof.
In some embodiments, the plant or plant part expresses a polypeptide that
comprises, consists essentially of or
consists of a fragment of one of SEQ ID NOs: 1-48 and 98-150 or a mature
peptide thereof. For example, the
microorganism may express a fragment of any one of SEQ ID NOs: 1-48 and 98-150
or a mature peptide thereof, said
fragment comprising at least 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
97, 98 or 99 % of the amino acids found in the
original protein.
In some embodiments, the plant or plant part expresses a polypeptide that
comprises, consists essentially of or
consists of any one of SEQ ID NOs: 1-48 and 98-150 or a mature polypeptide
thereof with an N-terminal extension of
one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,
15, 16, 17, 18, 19, 20 or more amino acids), a C-
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terminal extension of one or more amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8,
9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
more amino acids), one or more substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or
more substitutions), one or more insertions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
insertions), and/or one or more deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more
deletions). The amino acid changes may be of a minor nature, that is
conservative amino acid substitutions or insertions
that do not significantly affect the folding and/or activity of the protein:
small deletions, typically of 1-30 amino acids;
small amino- or carboxyl-terminal extensions, such as an amino-terminal
methionine residue; a small linker peptide of
up to 20-25 residues; or a small extension that facilitates purification by
changing net charge or another function, such as
a poly-histidine tract, an antigenic epitope or a binding module.
In some embodiments, the plant or plant part comprises a homologous or
heterologous nucleic acid sequence
that is at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the nucleic acid sequences set forth
herein as SEQ ID Nos: 49-97 and 151-203 or the cDNA sequence thereof.
In preferred embodiments, the plant or plant part comprises a polynucleotide
encodes a polypeptide that comprises,
consists essentially of or consists of the amino acid sequence of any one of
SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof.
In some embodiments, the plant or plant part comprises a polynucleotide that
comprises, consists essentially of
or consists of a nucleic acid sequence encoding a polypeptide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,
89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 'A
sequence identity to any one or more of SEQ ID NOs: 1-48 and 98-150.
In some embodiments, the plant or plant part comprises a polynucleotide that
comprises, consists essentially of
or consists of a nucleic acid sequence encoding a polypeptide having about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69,
70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,
89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to a mature polypeptide of any one or more of SEQ ID NOs: 1-
48 and 98-150.
In preferred embodiments, the plant or plant part comprises a polynucleotide
that comprises, consists essentially
of or consists of the nucleic acid sequence of any one of SEQ ID NOs: 49-97
and 151-203 or a cDNA sequence thereof.
In some embodiments, the plant or plant part comprises a polynucleotide that
encodes a polypeptide that
comprises, consists essentially of or consists of a fragment of one of SEQ ID
NOs: 1-48 and 98-150 or a mature peptide
thereof. For example, the microorganism comprises a polynucleotide that
encodes a fragment of any one of SEQ ID
NOs: 1-48 and 98-150 or a mature peptide thereof, said fragment comprising at
least 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98 or 99 % of the amino acids found in the original protein.
In some embodiments, the plant or plant part comprises a polynucleotide that
encodes a polypeptide that
comprises, consists essentially of or consists of any one of SEQ ID NOs: 1-48
and 98-150 or a mature polypeptide
thereof with an N-terminal extension of one or more amino acids (e.g., 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20 or more amino acids), a C-terminal extension of one or more
amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids), one or more
substitutions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19, 20 or more substitutions), one or more
insertions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19,20 or more insertions), and/or one or more
deletions (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20 or more deletions). The amino acid changes may
be of a minor nature, that is conservative
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amino acid substitutions or insertions that do not significantly affect the
folding and/or activity of the protein; small
deletions, typically of 1-30 amino acids; small amino- or carboxyl-terminal
extensions, such as an amino-terminal
methionine residue; a small linker peptide of up to 20-25 residues; or a small
extension that facilitates purification by
changing net charge or another function, such as a poly-histidine tract, an
antigenic epitope or a binding module.
The present disclosure also extends to plants, e.g., a transgenic plant, plant
part, or plant cell, comprising a
polynucleotide of the present disclosure so as to express and produce a
protein of the present disclosure in recoverable
quantities. The protein may be recovered fmm the plant or plant part.
Alternatively, the plant or plant part containing the
protein may be used as such for improving the quality of a food or feed, e.g.,
improving nutritional value, palatability, and
rheological properties, or to destroy an antinutritive factor.
The transgenic plant can be dicotyledonous (a dicot) or monocotyledonous (a
monocot). Examples of monocot
plants are grasses, such as meadow grass (blue grass, Poa), forage grass such
as Festuca, Lolium, temperate grass, such as
Agrostis, and cereals, e.g., wheat, oats, rye, barley, rice, sorghum, and
maize (corn).
Examples of dicot plants are tobacco, legumes, such as lupins, potato, sugar
beet, pea, bean and soybean, and
cruciferous plants (family Brassicaceae), such as cauliflower, rape seed, and
the closely related model organism
Arabidopsis thaliana.
Examples of plant parts are stem, callus, leaves, root, fruits, seeds, and
tubers as well as the individual tissues
comprising these parts, e.g., epidermis, mesophyll, parenchyme, vascular
tissues, meristems. Specific plant cell
compartments, such as chloroplasts, apoplasts, mitochondria, vacuoles,
peroxisomes and cytoplasm are also considered to
be a plant part. Furthermore, any plant cell, whatever the tissue origin, is
considered to be a plant part. Likewise, plant
parts such as specific tissues and cells isolated to facilitate the
utilization of the invention are also considered plant parts,
e.g., embryos, endospenns, aleurone and seed coats.
Also included within the scope of the present disclosure arc the progeny of
such plants, plant parts, and plant cells.
The transgenic plant or plant cell expressing the protein may be constructed
in accordance with methods known
in the art. In short, the plant or plant cell is constructed by incorporating
one or more expression constructs encoding the
protein into the plant host genome or chloroplast genome and propagating the
resulting modified plant or plant cell into a
transgenic plant or plant cell. In an embodiment, a plant cell does not belong
to plant varieties.
The expression construct is conveniently a nucleic acid construct that
comprises a polynucleotide encoding a
protein, wherein the polynucleotide is operably linked with appropriate
regulatory sequences required for expression of
the polynucleotide in the plant or plant part of choice. Furthermore, the
expression construct may comprise a selectable
marker useful for identifying plant cells into which the expression construct
has been integrated and DNA sequences
necessary for introduction of the construct into the plant in question (the
latter depends on the DNA introduction method
to be used).
Thc choicc of regulatory sequences, such as promoter and terminator sequences
and optionally signal or transit
sequences, is determined, for example, on the basis of when, where, and how
the protein is desired to be expressed (Sticklen,
2008, Nature Reviews 9: 433-443). For instance, the expression of the gene
encoding a protein may be constitutive or
inducible, or may be developmental, stage or tissue specific, and the gene
product may be targeted to a specific tissue or
plant part such as seeds or leaves. Regulatory sequences are, for example,
described by Tague et al., 1988, Plant Physiology
86: 506.
For constitutive expression, the 35S-CaMV, the maize ubiquitin 1, or the rice
actin 1 promoter may be used
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(Franck et at., 1980, Cell 21: 285-294; Christensen et al., 1992, Plant Mol.
Biol. 18: 675-689; Zhang et al., 1991, Plant
Cell 3: 1155-1165). Organ-specific promoters may be, for example, a promoter
from storage sink tissues such as seeds,
potato tubers, and fruits (Edwards and Coruzzi, 1990, Ann. Rev. Genet. 24: 275-
303), or from metabolic sink tissues such
as meristems (Ito et at., 1994, Plant Alol. Biol. 24: 863-878), a seed
specific promoter such as the glutelin, prolamin,
globulin, or albumin promoter from rice (Wu et al., 1998, Plant Cell Physiol.
39: 885-889), a Vicia ,faba promoter from
the legumin B4 and the unknown seed protein gene from Vicia faba (Conrad et
al., 1998, J. Plant Physiol. 152: 708-711),
a promoter from a seed oil body protein (Chen et al., 1998, Plant Cell
Physiol. 39: 935-941), the storage pmtein napA
promoter from Brass/ca napus, or any other seed specific promoter known in the
art, e.g., as described in WO 91/14772.
Furthermore, the promoter may be a leaf specific promoter such as the rbcs
promoter from rice or tomato (Kyozuka et al.,
1993, Plant Physiol. 102: 991-1000), the chlorella virus adenine
methyltransferase gene promoter (Mitra and Higgins,
1994, Plant Mol. Biol. 26: 85-93), the aldP gene promoter from rice (Kagaya et
al., 1995, Mol. Gen. Genet. 248: 668-674),
or a wound inducible promoter such as the potato pin2 promoter (Xu et al.,
1993, Plant Mol. Biol. 22: 573-588). Likewise,
the promoter may be induced by abiotic treatments such as temperature,
drought, or alterations in salinity or induced by
exogenously applied substances that activate the promoter, e.g., ethanol,
oestrogens, plant hormones such as ethylene,
abscisic acid, and gibbercllic acid, and heavy metals.
A promoter enhancer element may also be used to achieve higher expression of a
protein in the plant. For instance,
the promoter enhancer element may be an intron that is placed between the
promoter and the polynucleotide encoding a
protein. For instance, Xu et al., 1993, supra, disclose the use of the first
intron of the rice actin 1 gene to enhance expression.
The selectable marker gene and any other parts of the expression construct may
be chosen from those available
in the art.
The nucleic acid construct is incorporated into the plant genome according to
conventional techniques known in
the art, including Agrobacterium-mediated transformation, virus-mediated
transformation, microinjection, particle
bombardment, biolistic transformation, and electroporation (Gasser et at.,
1990, Science 244: 1293; Potrykus, 1990,
Bio/Technologv 8: 535; Shimamoto et al., 1989, Nature 338: 274).
Agrobacterium tumefaciens-mediated gene transfer is a method for generating
transgenic dicots (for a review, see
Hooykas and Schilperoort, 1992, Plant Mot Biol. 19: 15-38) and for
transforming monocots, although other transformation
methods may be used for these plants. A method for generating transgenic
monocots is particle bombardment (microscopic
gold or tungsten particles coated with the transforming DNA) of embryonic
calli or developing embryos (Christou, 1992,
Plant 1 2: 275-281; Shimamoto, 1994, Curr. Op/n. Biotechnol. 5: 158-162; Vasil
et al., 1992, Bio/Technology 10: 667-
674). An alternative method for transformation of monocots is based on
protoplast transformation as described by
Omirulleh et al., 1993, Plant Mol. Biol. 21: 415-428. Additional
transformation methods include those described in U.S.
Patent Nos. 6,395,966 and 7,151,204 (both of which are herein incorporated by
reference in their entirety).
Following transformation, thc transformants having incorporated thc expression
construct arc selected and
regenerated into whole plants according to methods well known in the art.
Often the transformation procedure is designed
for the selective elimination of selection genes either during regeneration or
in the following generations by using, for
example, co-transformation with two separate T-DNA constructs or site-specific
excision of the selection gene by a specific
recombinase.
In addition to direct transformation of a particular plant genotype with a
construct of the present disclosure,
transgenic plants may be made by crossing a plant having the construct to a
second plant lacking the construct. For example,
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a construct encoding a protein can be introduced into a particular plant
variety by crossing, without the need for ever
directly transforming a plant of that given variety. Therefore, the present
disclosure encompasses not only a plant directly
regenerated from cells which have been transformed in accordance with the
present disclosure, but also the progeny of
such plants. As used herein, progeny may refer to the offspring of any
generation of a parent plant prepared in accordance
with the present disclosure. Such progeny may include a DNA construct prepared
in accordance with the present disclosure.
Crossing results in the introduction of a transgene into a plant line by cross
pollinating a starting line with a donor plant
line. Non-limiting examples of such steps are described in U.S. Patent No.
7,151,204.
Plants may be generated through a process of backcross conversion. For
example, plants include plants referred
to as a backcross converted genotype, line, inbred, or hybrid.
Genetic markers may be used to assist in the introgression of one or more
transgenes of the disclosure from one
genetic background into another. Marker assisted selection offers advantages
relative to conventional breeding in that it
can be used to avoid errors caused by phenotypic variations. Further, genetic
markers may provide data regarding the
relative degree of elite germplasm in the individual progeny of a particular
cross. For example, when a plant with a desired
trait which otherwise has a non-agronomically desirable genetic background is
crossed to an elite parent, genetic markers
may be used to select progeny which not only possess the trait of interest,
but also have a relatively large proportion of the
desired germplasm. In this way, the number of generations required to
introgress one or more traits into a particular genetic
background is minimized.
The present disclosure also relates to methods of producing a protein of the
present disclosure comprising (a)
cultivating a transgenic plant or a plant cell comprising a polynucleotide
encoding the protein under conditions conducive
for production of the protein; and (b) recovering the protein.
The present disclosure also extends to methods of producing a mutant of a
parent cell, which comprises disrupting
or deleting a polynucleotide, or a portion thereof, encoding a protein of the
present disclosure, which results in the mutant
cell producing less of the protein than the parent cell when cultivated under
the same conditions.
The mutant cell may be constructed by reducing or eliminating expression of
the polynucleotide using methods
well known in the art, for example, one or more nucleotide insertions, one or
more gene disruptions, one or more nucleotide
replacements, or one or more nucleotide deletions.
The polynucleotide to be modified or inactivated may be, for example, the
coding region or a part thereof essential
for activity, or a regulatory or control element required for expression of
the coding region, e.g.. a functional part of a
promoter sequence, and/or a regulatory or control element required for the
transcription or translation of the polynucleotide.
Other control sequences for possible modification include, but are not limited
to, a leader, polyadenylation sequence,
propeptide sequence, signal peptide sequence, transcription terminator, and
transcriptional activator.
Modification or inactivation of the polynucleotide may be performed by
subjecting the parent cell to mutagenesis
and selecting for mutant cells in which expression of the polynucleotide has
bccn reduced or eliminated. Thc mutagenesis,
which may be specific or random, may be performed, for example, by use of a
suitable physical or chemical mutagenizing
agent, by use of a suitable oligonucleotide, or by subjecting the DNA sequence
to PCR generated mutagenesis. Furthermore,
the mutagenesis may be performed by use of any combination of these
mutagenizing agents.
Examples of a physical or chemical mutagenizing agent include ultraviolet (UV)
irradiation, hydroxylamine,
N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 0-methyl hydro,xylamine, nitrous
acid, ethyl methane sulphonate (EMS),
sodium bisulphite, formic acid, and nucleotide analogues (see J. L. Bose,
Springer Protocols 2016, Methods in _Molecular
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Biology, The Genetic Manipulation of Staphylococci).
Additionally or alternatively, nucleotides may be inserted or removed so as to
result in the introduction of a stop
codon, the removal of the start codon, or a change in the open reading frame.
Such modification or inactivation may be
accomplished by site-directed mutagenesis or PCR generated mutagenesis in
accordance with methods known in the art,
or by targeted gene editing using one or more nucleases, e.g., zinc-finger
nucleases or CRISPR-associated nucleases.
Additionally or alternatively, the modification or inactivation may be
achieved by gene silencing, genetic repression,
genetic activation, and/or post-translational mutagenesis, e.g., by methods
employing non-coding RNA, RNAi, siRNA,
miRNA, ribozymes, catalytically inactive nucleases, CRISPRi, nucleotide
methylation, and/or histone acetylation. The
modification may be transient and/or reversible, irreversible and/or stable,
or the modification may be dependent on
chemical inducers or dependent on cultivation conditions, such as the
cultivation temperature.
The modification may be performed in vivo, i.e., directly on the cell
expressing the polynucleotide to be modified,
or the modification be performed in vitro.
An example of a convenient way to eliminate or reduce expression of a
polynucleotide is based on techniques of
gene replacement, gene deletion, or gene disruption. For example, in the gene
disruption method, a nucleic acid sequence
corresponding to the endogenous polynucleotide is mutagcnized in vitro to
produce a defective nucleic acid sequence that
is then transformed into the parent cell to produce a defective gene. By
homologous recombination, the defective nucleic
acid sequence replaces the endogenous polynucleotide. It may be desirable that
the defective polynucleotide also encodes
a marker that may be used for selection of transformants in which the
polynucleotide has been modified or destroyed. In
an aspect, the polynucleotide is disrupted with a selectable marker such as
those described herein.
The present disclosure further relates to a mutant cell of a parent cell that
comprises a disruption or deletion of a
polynucleotide encoding the protein or a control sequence thereof or a
silenced gene encoding the protein, which results
in the mutant cell producing less of the protein or no protein compared to the
parent cell.
The protein-deficient mutant cells are useful as host cells for expression of
native and heterologous proteins.
Therefore, the present disclosure further relates to methods of producing a
native or heterologous protein, comprising (a)
cultivating the mutant cell under conditions conducive for production of the
protein; and (b) recovering the protein. The
term "heterologous proteins" means proteins that are not native to the host
cell, e.g., a variant of a native protein. The host
cell may comprise more than one copy of a polynucleotide encoding the native
or heterologous protein.
The methods of the present disclosure for producing an essentially [enzymel-
free product are of interest in the
production of proteins, e.g., proteins such as enzymes. The [enzymel-deficient
cells may also be used to express
heterologous proteins of pharmaceutical interest such as hormones, growth
factors, receptors, and the like.
In some embodiments, the present disclosure relates to a protein product
essentially free from [enzyme] activity
that is produced by a method of the present disclosure.
Compositions of the present disclosure arc useful for preventing, treating,
suppressing, eliminating and/or
reducing the severity of infestations/infections of/by myriad pests,
including, but not limited to, horticultural pests, such
as acarids, bacteria, fungi, gastropods, insects, nematodes, oomycetes,
protozoa and weeds.
Compositions of the present disclosure (i.e., proteins, formulations,
polynucleotides and organisms of the
present disclosure) may be used to prevent, treat, suppress, eliminate and/or
reduce the severity of acarid infestations by,
for example, reducing attraction of an acarid to a surface, inhibiting entry
of an acarid into a material, inhibiting
inhabitation by an acarid, inhibiting feeding of an acarid, inhibiting the
growth of an acarid, inhibiting the reproduction
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and/or proliferation of an acarid, degrading on one or more structural
components of an acarid (e.g. procuticle
components, such as chitin, and epicuticle components, such as lipoproteins,
hydrocarbons, polyphenols and esters of
fatty acids and alcohols), killing an acarid, and/or reducing one or more
symptoms of an acarid infestation. In some
embodiments, such inhibition is complete or substantially complete, such that
the acarid fails to
inhabit/feed/grow/reproduce/proliferate at a rate effective to initiate and/or
sustain an appreciable infestation. For
example, spraying a plant with an enzyme of the present disclosure may reduce
the attractiveness of the plant to an
acarid, reduce an acarid's desire/ability to inhabit the plant, inhibit an
acarid's desire/ability to feed on the plant,
inhibiting the growth of an acarid after it inhabits/feeds on the plant,
inhibit the reproduction and/or proliferation of an
acarid on/in the plant, degrade one or more structural components of an acarid
(e.g. one or more chitins, one or more
lipoproteins and/or one or more long chain hydrocarbons) on/in the plant,
and/or kill an acarid on/in the plant, thereby
reducing one or more symptoms of infestation and/or enhancing one or more
characteristics of growth and/or yield in the
plant, as compared to an untreated control plant.
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of infestations of/by myriad acarids, including, but not limited to,
herbivorous acarids, such as broad mites, bulb
mites, cyclamen mites, earth mites, criophyid mites, Lewis mites, russet
mites, rust mites, and spider mites. In some
embodiments, enzymes of the present disclosure are used to prevent and/or
treat an infestation of a plant or plant part by
one or more Auculops, (e.g., A. lycopersici), Calacarus (e.g., C. f lagelli
seta), Eutetranychus (e.g., E. lewesi), Halotydeus
(e.g., H. destructor), Polyphagotarsonemus (e.g., P. latus), Rhizoglyphus,
Tarsonemus (e.g., T. palhdus), and/or
Tetranychus (e.g., T. cinnabarinus, T. evansi, T. ludeni, T. urticae).
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of bacterial infestations/infections by, for example, inhibiting
adhesion of a bacterium to a surface, inhibiting
entry of a bacterium into a material, inhibiting habitation by a bacterium,
inhibiting the growth of a bacterium, inhibiting
the reproduction and/or proliferation of a bacterium, degrading on one or more
structural components of a bacterium
(e.g. cell wall components, such as peptidoglycans and lipopolysaccharides),
killing a bacterium, and/or reducing one or
more symptoms of a bacterial infestation/infection. In some embodiments, such
inhibition is complete or substantially
complete, such that the bacterium fails to
inhabit/feed/grow/reproduce/proliferate at a rate effective to initiate and/or
sustain an appreciable infestation/infection. For example, spraying a plant
with an enzyme of the present disclosure may
inhibit a bacterium's ability to adhere to the surface of a plant, inhibit a
bacterium's ability to enter into the plant, inhibit a
bacterium's ability to inhabit the plant, inhibit growth of a bacterium on/in
the plant, inhibit the reproduction and/or
proliferation of a bacterium on/in the plant, degrade one or more structural
components of a bacterium (e.g. one or more
peptidoglycans and/or one or more lipopolysaccharides) on/in the plant, and/or
kill a bacterium, thereby reducing one or
more symptoms of infestation and/or enhancing one or more characteristics of
growth and/or yield in the plant, as
compared to an untreated control plant.
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of infestations/infections of/by myriad phytopathogenic bacteria,
including, but not limited to, phytopathogenic
Erwiniaceae and Xanthomonadales. In some embodiments, enzymes of the present
disclosure are used to prevent, treat,
suppress, eliminate and/or reduce the severity of an infestation/infection of
a plant or plant part by one or more
Agrobacterium (e.g., .4. rhizogenes, A. tutnefaciens, A. vitis), Burkholderia
(e.g., B. gladioli), Clostridium, Dickeyet (e.g.,
D. dadantii, D. solani),Erwinia (e.g., E amylovora, F. aphidicola, F.
carolovora, E. chrysanthemi, E papayne, F.
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persicina, E. psidii, E. pyrijbliae, E. rhapontici, E. tracheiphila),
Pectobacterium (e.g., P. atrosepticum, P.
carotovorum), Pseudomonas (e.g., P. agarici, P. amygdah, P. avellanae,P.
cannabina, P. caricapapayae, P. cichorii, P.
coronajaciens, P. co,stantinii, P. ficuserectae, P. fitscovaginae, P.
hellanthin, P. mellae, P. ,savastanoi, P. ,syringae, P.
tolaasii, P. tomato, P. turbinellae, P. viridiflava), Ralstonia (e.g., R.
solanacearum), Xanthomonas (e.g., X alfalfae, X
ampelina, X arbori cola, X axonopodia, X boreopolis, X badrii, X bromi, X
campestris, X cassavae, X citri, X
cucrurbitae, X. cyanopsidis, X cynarae, Xeuvesicatoria , X. frageriae,
Xgardneri, X holcicola, X hortorum, X
hyacinthi, X maliensis, X malvacearum, X tnanihotis, X tnelonis, X oryzae, X
papavericola, X perforans, X phaseoh,
X pisi, X populi, X sacchari, X the/cola, X translucens, X vas/cola, X
vesicatoria), and/orXy/ella (e.g., X fastidiosa).
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of fungal infestations/infections by, for example, inhibiting
adhesion of a fungus to a surface, inhibiting entry of
a fungus into a material, inhibiting habitation by a fungus, inhibiting the
growth of a fungus, inhibiting the reproduction
and/or proliferation of a fungus, degrading on one or more structural
components of a fungus (e.g. cell wall components,
such as chitins, glucans and mannans, and cell membrane components, such as
ergosterols), killing a fungus, and/or
reducing one or more symptoms of a fungal infestation/infection. In some
embodiments, such inhibition is complete or
substantially complete, such that the fungus fails to
inhabit/feed/grow/reproduce/proliferate at a rate effective to initiate
and/or sustain an appreciable infestation/infection. For example, spraying a
plant with an enzyme of the present
disclosure may inhibit a fungus' ability to adhere to the surface of a plant,
inhibit a fungus' ability to enter into the plant,
inhibit a fungus' ability to inhabit the plant, inhibit growth of a fungus
on/in the plant, inhibit the reproduction and/or
proliferation of a fungus on/in the plant, degrade one or more structural
components of a fungus (e.g. one or more
chitins, one or more glucans and/or one or more mannans) on/in the plant,
and/or kill a fungus, thereby reducing one or
more symptoms of infestation and/or enhancing one or more characteristics of
growth and/or yield in the plant, as
compared to an untreated control plant.
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of infestations/infections of/by myriad phytopathogcnic fungi,
including, but not limited to, phytopathogcnic
Ascomycetes, Basidiomycetes, Chytridiomycota, Deueromycota, Peronosporomycota,
Plasmocliophoromycota and
Zygomycota, such as blasts, blights, bunts, galls, mildews, molds, rots,
rusts, scabs, smuts and wilts. In some
embodiments, enzymes of the present disclosure are used to prevent, treat,
suppress, eliminate and/or reduce the severity
of an infestation/infection of a plant or plant part by one or more Aecidium
(e.g., A. aechmantherae, A. amarylhdis, A.
breyniae, A. campanulastri, A. cannabis, A. cantensis, A. capsicum, A.
foeniculi, A. narcissi), Alternaria (e.g., A.
alternata, A. alternantherae, A. arachidi,s, A. arbore,scen,s, A. arbliSii, A.
blumeae, A. brassicae, A. brassicicola, A.
burns/i. A. carotiincultae, A. carthami, A. celosiae, A. cinerariae, A. citri,
A. conjuncta, A. cucumerina, A. dauci, A.
dianthi, A. dianthicola, A. eichhorniae, A. euphorbiicola, A. gaisen, A.
helianthi, A. helianthi cola, A. hungarica, A.
infector/a, A. japonica, A. leucanthemi, A. hnicola, A. longipes, A. mall, A.
molesta, A. padivickii, A. pa/lax, A.
perpunctulata, A. patrosehni, A. porn, A. queri cola, A. radicina, A. raphani,
A. saponariae, A. A. senecionis, A.
smyrnii, A. solani, A. sonchi, A. tenuissima, A. triticina, A. ventricosa, A.
zinniae), Ascochyta (e.g., A. asparagina, A.
bohemica, A., caricae, A. doronici, A. fabae, A. gossypii, A. graminea, A.
horde/, A. humuli, A. medicaginicola, A.
pinodes, A. pisi, A. prasadii, A. rabiei, A. rhei, A. sorghi, A. sorghinia, A.
spinaciae, A. tarda, A. tritici, A. viciae, A.
vindobonensis), Ascospora (e.g., A. ruborum),Aspergillus (e.g., A.
actileattis, A. candidus, A. clavatus, A. fisher/anus, A.
jlavus, A. fitmigatus, A. niger, A. parasiticus, A. restricius, sojae, A.
solani), Asteroma (e.g., A. caryae),
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Austropuccinia (e.g., A. psidii), Bipolaris (e.g., B. cactivora, B. cookei, B.
incurvata, B. sacchari, B. sorghi cola, B. zeae),
Blumeria (e.g., B. graminis), Boeremia (e.g., B. lycopersici),Botrytis (e.g.,
B. al/ii, B. anthophila, B. cinerea, B.
caricola, B. citrina, B. elliptica, B. Jaime, B. jablop,si,s, B. galanthina,
B. gladioli, B. gossypina, B. hormini, B. hyacinthi,
B. isabellina, B. latebri cola, B. liliorum, B. limacidae, B. luteobrunnect,
B. lutescens, B. mali, B. monilioides, B.
narcissicola, B. necans, B. paeoniae, B. peronosporoides, B. pistiae, B.
platensis, B. pruinosa, B. pseudocinerea, B.
pyramidalis, B. rivoltae, B. rosea, B. rub escens, B. rudiculoides, B.
sekimotoi, B. septospora, B. setuligera, B. sinoallii,
B. sonatina, B. splendida, B. squamosa, B. taxi, B. terrestris, B.
tracheiphila, B. trifolii, B. tulipae, B. viciae-hirsutae, B.
yucte), Calonectria (e.g., C. ilicicola, C. inc/us/ate, C. kyotensis, C.
pteridis, C. pyrochroa, C. quinqueseptata),
Camarotella (e.g., C. acrocomiae, C. costaricensis), Candida (e.g., C.
albicans), Capnodium (e.g., C. theae),
Cephalosporium (e.g., C'. gramineum), Ceratocystis (e.g., C. Jimbriata),
Ceratobasidium (e.g., (J'. cereale),
Cercoseptoria (e.g., C. ocellata), Cercospora (e.g., C. angreci, C. apii, C.
apiicolct, C. arachidicola, C. asparctgi, C.
airofiliformis, C. beticola, C. bolleana, C. brachypits, C. brassicola, C
brunkii, C. canescens, C. cannabis, C.
cantuariensis, C. capsici, C. caribaea, C. carotae, C. circumscissa, C.
citrulline, C. clemensiae, C. coffeicola, C. coryli,
C. corylina, C. eleusine, C. fragariae, C. fuchsiae, Cjusca, Cjusimaculans, C.
gerberae, C. halstedii, C. handelii, C.
hayi, C. hydrangeae, C. kaki, C. kikuchii, C. lentis, C. liquidambraris, C.
longipes, C. longissima, C. malloti, C.
mamaonis, C. mangiferae, C. medicaginis, C. melongenae, C. minima, C. minuta,
C. musae, C. nicotianae, C.
odontoglossi, C. oryzae, C. papayae, C. penniseti, C. personata, C. pictropi,
C. pisa-sativae, C. platanicola, C. puderii,
C. pulcherrima, C. rhaptchcola, C. rostcola, C. rubrotincta, C. sopna, C.
so/ant, C. solani-tuberost, C. sorght, C. theae,
C. tuberculans, C. vexans, C. vicosae, C. zeae-maydis, C. zebrina, C. zonata),
Cercosporella (e.g., C. rubi),
Choanephora (e.g., C. cucurbilarum), Cladosporium (e.g., C. arthropodii, C.
brassicae, C. brass/cola, C. chrysanthemi,
C. cari, C. cladasporioide,s, C. cucumerinum, C. fulvum, C. gossyplicola, C.
herbarum, C. hydrangeae, C. leguminicola,
C. musae, C. oncobae, C. orchid/s, C. pisi, C. rhododendri, C. salinae, C.
spharospermum, C. sorghi, C. syringae, C.
syringicola, C. yuccae, C. zeae), Claviceps (e.g., C. nfricana,C..fusiformis,
C. paspali, C. purpurea, C. sorghi, C.
zizaniae), Clitocybe (e.g., C. parasitica), Coccidioides (e.g., C. immitus),
Cochliobolus (e.g., C. carbonum, C.
cymbopogonis, C. hawallensis, C. heterostrophus, C. lzinatus, C. miyabeanus,
C. ravenelii, C. sativus, C. setariae, C.
spicifer, C. stenospilus, C. tuberculatus, C. victoriae), Coleasporium (e.g.,
C helianthi, C. ipomoene, C marline, C.
pacificum, C. tussilaginis), Colletotrichum (e.g., C. acutatum, C. agaves, C.
arachidis, C. boninense, C. brasihense, C.
brass/co/a, C. brevisporum, C. cacao, C. capsici, C. caudatum, C. cereale, C.
citri, C. citricola, C. coccodes, C.
coffeanum, C. crassipes, C. curcumae, C. dematium, C. derridis, C.
destructivum, C. form/ac, C. fragariae, C. fructi, C.
jruticola, C jructivorum, C gloeasporioldes, C glycines, (1 gram inicola, C.
gossypii, C. hanaul, C higginsianum, C
jacksonii, C. kahawae, C. lentis, C. limonicola, C. lindemuthianum, C. lini,
C. lupin', C. mangenotii, C. melonis, C.
miscanthi, C. musae, C. nicholsonii, C. nigrum, C. orb/cu/are, C. orchidis, C.
paspali, C. pisi, C. pis/cola, C. radicis, C.
roseum, C. serranegrense, C. sojae, C. spinaceae, C. sub lineolum, C. sub
lineola, C. tab acum, C. trichellum, C. trifolii,
C. truncatum, C. zoysiae),Coniella, Coniothecium (e.g., C.
chomatosporum),Coniothyrium (e.g., C. henriquesii, C.
rosarum, C. wernsdorffiae), Coprinopsis (e.g., C. p.sychromorbida), Cordana
(e.g., C. johnstonii, C. musae), Corticum
(e.g., C. theae), Cryphonectria (e.g., C. parasitica), Cyhndrocarpon (e.g., C.
ianthothele, C. magnusianum, C. musae),
Cylindrocladiella (e.g., C. camel//ac, C. parva), Cylindrocladium (e.g., C.
lanceolatum, C. peruvianum),
Cylindrosporium (e.g., C. cannabinum, C. jzzglandis, C. rubi), Cvmadothea
(e.g., C. trifolii), Cvtospora (e.g, C.
palmarum, C. persona/a, C sacchari, C ATICC111 1 IS, C. terebinthi),
Cyto.sporina (e.g., C. ludibunda),Diaporthe (e.g., D.
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arctii, D. asparagi, D. capsici, 1). citri, D. coffeae, D. dulcamarae, D.
eres, D. helianthi, 1). lagunensis, D. lokoyae, D.
melonis, D. musae, D. orthoceras, D. perniciosa, D. phaseolorum, D. rudis, D.
tanakae, D. viticola), Diplodia (e.g., D.
gossypina), Dre,vchlera (e.g., D. avenacea, 1). campanulata, 1). dematioidea,
1). gigantea, 1). g1vcine,v, D. gram inea, 1).
hawaiiensis, D. musae, D. poae, D. teres, D. ivirreganensis), Eremothcium
(formerly Nematospora) (e.g., E. gossypii),
Erysiphe (e.g., E. betae, E. cichoracearum, E. communis, E. cruciferarum, E.
llexuosa, E. heraclei, E. necator, E. pisi, E.
polygoni, E. robiniae, E. syringae), Exserohilum (e.g., E. oryzicola, E.
oryzinum), Fusarium (e.g., F. affine, F.
arthrosporioides, F. avenaceum, F. circinatum, F. crookwellense, F. culmorum,
F. fujikuroi, F. graminearutn, F.
incarnatum, F. langsethiae, F. mangtferae, F. merismoides, F. moniliforme, F.
oxysporum, F. pallidoroseum, F. poae, F.
proltferatum, F. redo/ens. F. roseum, F. sacchari, F. solani, F.
sporotrichioides, F. sterilihyphosum, F. subglutinans, F.
sulphureum, F tricinctum, F. verticillioides, F. virgulifOrme), Gaeumannomyces
(e.g., G. graminis), Geotrichum (e.g.,
G. candidum), Gibberella (e.g., G. fujikuroi, G. pulicaris, G. stilboides, G.
tricincta, G. xylarioides, G. zeae), Gilbertella
(e.g., G. pervicaria), Glomerella (e.g., G. eingulata), Gymno.sporangium
(e.g., G. juniperi-virginianae),
Helminthosporium (e.g., If. oryzae, If. solani), Hemileia (e.g., IL
coffeicola, vastatrix), Leptosphaeria (e.g., L. acuta,
L. asparagi, L. cannabina, L. coffaeicida, L. coniothyrium, L. glweriae, L.
gossypii, L. grisea, L. longispora, L.
maculans, L. maydis, L. musae, L. oryzicola, L. oryzina, L. pint, L.
platanicola, L. pratensis, L. raphani, L. saccharicola,
L. solani, L. solanicola, L. trtfblii, L. viciae, L. woroninii, L. zeae, L.
zeae-maydis), Macrophomina (e.g., M phaseolina),
Magnaporthe (e.g., M. grise a, M oryzae), Melamspora (e.g., NI lint),
11/fonilinia (e.g., M. fructicola), Mucor (e.g., 111.
',informs), Mycosphaerella (e.g., M. fulensis, M. grammicola, Al tassiana, NI.
zeae-maydis), Neofabraea (e.g., N.
malicorticus), Ophiostoma (e.g., 0. novo-ulmi, 0. ulmi), Paracoccidioides
(e.g., P. braziliensis), Penicillium (e.g., P.
digitalum, P. expansum, P. italicum, P. rugulosum, P. verrucosum), Phakopsora
(e.g., P. gossypii, P. meibomiae, P.
pachyrhizi), Phialophora (e.g., P. gregata), Phoma (e.g., P. glycinicola),
Phomop,vis (e.g., P. asparagi, P. coffeae, P.
logicolla, P. mangiferae, P. obscurans, P. perseae, P. purnorum, P. sojae, P.
sclerotioides, P. tan akae, P. theae, P.
vitcola), Phymatotrichopsis (e.g., P. omnivora), Physalospora (e.g., P.
obtusa), Phytomyxea, Pneumocystis (e.g., P.
carinii), Podosphaera (e.g., P. oxyacanthae), Pseudocercosporella, Puccinia
(e.g. , P. asparagi, P. cacahata, P.
graminis, P. kuehnii, P. melanocephala, P. porn, P. punctiforrnis, P.
recondita, P. schedonnardii, P. sessilis, P. sorghi,
P. striiformis, P. tritici, P. triticina), Pyrenophora (e.g., P. tritici-
repentis), Pyricularia, Rhizoctonia (e.g., R. solani),
Rhizopus (e.g., R. nigri cans, R. stolonifer), Rhynchosporium, Sclerotinia
(e.g., S. borealis, S. bulborum, S. homoeocarpa,
S. libertiana, S. minor, S. ricini, S. sclerotiorum, S. spermophila, S.
trilbliorum), Sclerotium (e.g., S. rollsii),
Scopulariopsis (e.g., S. brevicaulis), Septoria (e.g., S. apiicola, S.
aciculosa, S. arnpelina, S. avenae, S. azalea, S.
bataticola, S1 campanulae, S1 caryae, 51 cari, 51 cucurbitacearum, S1 cytisi,
S1 &anal", 51 eumu,vae, Jragariae, 51
fragariaecola, S. glycines, S. helianthin, S. humuli, S. hydrangea, S.
lactucae, S. lycopersici, S. malagutii, S. menthae, S.
musiva, Si ostryae, Si passerinii, Si pisi, S. pistaciae, S platantfolia, Si
rhododendri, Si secais, S'. selenophomoides),
Sporisorium (e.g., S. scitamineum), Synch ytrium (e.g., S. endobioticum),
Taphrina (e.g., T, deformans), Thielaviopsis
(e.g., T. basicola, T. ceremica), Tilletia (e.g., T. barclaycma, T. caries, T.
contra versa, T. foetida, T. indica, T. laevis, T.
tritici), Uncinula, Uromyces (e.g., U melanocephala), Ustilago (e.g., U.
esculenta, U. maydis, U. nuda, U scitaminea,
tritici, U virens), Venturia,Verticillium (e.g., V. alfalfae, V dahliae, V.
isaacii, V longisporum, V. theobromae, V.
zaregamsianum) and/or Zymoseptoria (e.g., Z. tritici). Additional examples of
fungi that may be targeted using proteins,
formulations, polynucleotides and organisms of the present disclosure may be
found in Bradley, Managing Diseases, in
ILLINOIS AGRONOMY HANDBOOK (2008).
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Compositions of the present disclosure may be used to prevent and/or treat
gastropod infestations by, for
example, reducing attraction of a gastropod to a surface, inhibiting
inhabitation by a gastropod, inhibiting feeding of a
gastropod, inhibiting the growth of a gastropod, inhibiting the reproduction
and/or proliferation of a gastropod,
degrading on one or more structural components of a gastropod (e.g.
___________________ ), killing a gastropod, and/or reducing
one or more symptoms of a gastropod infestation. In some embodiments, such
inhibition is complete or substantially
complete, such that the gastropod fails to
inhabit/feed/grow/reproduce/proliferate at a rate effective to initiate and/or
sustain an appreciable infestation. For example, spraying a plant with an
enzyme of the present disclosure may reduce
the attractiveness of the plant to a gastropod, reduce a gastropod's
desire/ability to inhabit the plant, inhibit a gastropod's
desire/ability to feed on the plant, inhibiting the growth of a gastropod
after it inhabits/feeds on the plant, inhibit the
reproduction and/or proliferation of a gastropod on/in the plant, degrade one
or more structural components of a
gastropod (e.g.
_______________________________________________________________________ )
on/in the plant, and/or kill a gastropod, thereby reducing one or more
symptoms of infestation
and/or enhancing one or more characteristics of growth and/or yield in the
plant, as compared to an untreated control
plant.
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of infestations of/by myriad gastropods, including, but not limited
to, slugs and snails.
Compositions of the present disclosure may be used to prevent and/or treat
insect infestations by, for example,
reducing attraction of an insect to a surface, inhibiting entry of an insect
into a material, inhibiting inhabitation by an
insect, inhibiting feeding of an insect, inhibiting the growth of an insect,
inhibiting the reproduction and/or proliferation
of an insect, degrading on one or more structural components of an insect
(e.g. procuticle components, such as chitin,
and epicuticle components, such as lipoproteins, hydrocarbons, polyphenols and
esters of fatty acids and alcohols),
killing an insect, and/or reducing one or more symptoms of an insect
infestation. In some embodiments, such inhibition
is complete or substantially complete, such that the insect fails to
inhabit/feed/grow/reproduce/proliferate at a rate
effective to initiate and/or sustain an appreciable infestation. For example,
spraying a plant with an enzyme of the present
disclosure may reduce the attractiveness of the plant to an insect, reduce an
insect's desire/ability to inhabit the plant,
inhibit an insect's desire/ability to feed on the plant, inhibiting the growth
of an insect after it inhabits/feeds on the plant,
inhibit the reproduction and/or proliferation of an insect on/in the plant,
degrade one or more structural components of an
insect (e.g. one or more chitins, one or more lipoproteins and/or one or more
long chain hydrocarbons) on/in the plant,
and/or kill an insect, thereby reducing one or more symptoms of infestation
and/or enhancing one or more characteristics
of growth and/or yield in the plant, as compared to an untreated control
plant.
Compositions of the present disclosure may be used to prevent and/or treat
infestations of myriad insects,
including, but not limited to, Coleopteran Dermaptera, Diptera, Hemiptera,
Homoptera, Hymenoptera, Lepidoptera,
Orthoptera and Thysanoptera, such as aphids, armyworms, beetles, bollworms,
bugs, caterpillars, cutworms, flies, moths
and thrips. In somc embodiments, enzymes of the present disclosure arc used to
prevent, treat, suppress, eliminate and/or
reduce the severity of an infestation of by one or more Acalymma,
Acanthaoscelides (e.g., A. obtectus,), Anasa (e.g., A.
tristis), Anastrepha (e.g., A. ludens), Anoplophora (e.g., A. glabripennis),
Anthonomus (e.g., A. eugenii), Acyrthosiphon
(e.g., A. pisum), Bactrocera (e.g. B. dosalis), Bemisia (e.g., B.
argentifolii, B. tabaci), Brevicoryne (e.g., B. brassicae),
Bruchidius (e.g., B. atrolineatus), Bruchus (e.g., B. atomarius, B. dentipes,
B. lentis, B. pisorum and/or B. rufipes),
Callosobruchus (e.g., C. chinensis, C. maculatus, C. rhodesianus, C.
subinnotatus, C. theobromae), Caryedon (e.g., C.
yerralits), Cassadinae, Ceratilis (e.g., C capita/a), Chrysomelinde,
Circulifer (e.g., C it-menus); Criocerinae,
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C'ryptocephalinae, C'typtolestes (e.g., C. ferrugineus, C pusillis, C
puss/I/o/des), Cylas (e.g., C formicarius), Delia
(e.g., D. antiqua),Diabrotica,Diaphania (e.g., D. nitidalis), Diaphorina
(e.g., D. citri), Donaciinae, Ephestia (e.g, E.
cautella, E. elutella, E., keuhniella), Epilachna (e.g., E. varivestri,$),
Epiphyas (e.g., E. po,sivittana), Eumolpinae,
Galerucinae, Helicoverpa (e.g., H. zea), Heteroligus (e.g., H. me/es), Iobesia
(e.g., I. botrana), Lamprosomatinae,
Lasioderma (e.g., L. serricorne), Leptinotarsa (e.g., L. decemlineata),
Leptoglossus, Liriomyza (e.g., L. trifolii),
Manducca, Melittia (e.g., M. cucurbitae),Alyzus (e.g., M persicae), Nezara
(e.g., N. viridula), Orzaephilus (e.g., 0.
merator, 0. surinamensis), Ostrinia (e.g., 0. imbilalis), Phthorimaea (e.g.,
P. operculella), Piers (e.g., P. rapae), Plodia
(e.g., P. interpunctella), Plutella (e.g., P. xylostella), Pop/ilia (e.g., P.
japonica), Prostephanus (e.g., P. truncates), Ps/la,
Rhizopertha (e.g., R. dominica), Rhopalosiphum (e.g., R. maidis), Sagrinae,
Solenopsis (e.g., S. Invicta), Spilopyrinae,
Sitophilus (e.g., S. granaries, S. oryzae and/or S. zeamais), Sitotroga (e.g.,
S. cerealella), Spodoptera (e.g., S.
filigiperda), Stegobium (e.g., S. paniceum), Synetinae, Tenebrio (e.g., T.
ma/ens and/or T. molitor), Thrips (e.g., T.
labaci), Trialeurodes (e.g., T. vaporariorum), Tribolium (e.g., T. castaneum
and/or T. confitsum), Trichoplusia (e.g., T.
ni), Trogoderma (e.g., T. granarium) and/or Trogossitidae (e.g., T.
mauritanicus). Additional species of insects that may
be targeted using enzymes, formulations, polynucleotides and organisms of the
present disclosure may be found in
CAPINERA, HANDBOOK OF VEGETABLE PESTS (2001) and Steffcy and Gray, Managing
Insect Pests, in ILLINOIS
AGRONOMY HANDBOOK (2008).
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of nematode infestations/infections by, for example, inhibiting
adhesion of a nematode to a surface, inhibiting
entry of a nematode into a material, inhibiting habitation by a nematode,
inhibiting the growth of a nematode, inhibiting
the reproduction and/or proliferation of a nematode, degrading on one or more
structural components of a nematode (e.g.
cuticle components, such as collagens, cuticlins, glycoproteins and lipids),
killing a nematode, and/or reducing one or
more symptoms of a nematode infestation/infection. In some embodiments, such
inhibition is complete or substantially
complete, such that the nematode fails to
inhabit/feed/grow/reproduce/proliferate at a rate effective to initiate and/or
sustain an appreciable infestation/infection. For example, spraying a plant
with an enzyme of the present disclosure may
inhibit a nematode's ability to adhere to the surface of a plant, inhibit a
nematode's ability to enter into the plant, inhibit a
nematode's ability to inhabit the plant, inhibit growth of a nematode on/in
the plant, inhibit the reproduction and/or
proliferation of a nematode on/in the plant, degrade one or more structural
components of a nematode (e.g., one or more
collagens, one or more cuticlins, one or more glycoproteins and/or one or more
lipids) on/in the plant, and/or kill a
nematode, thereby reducing one or more symptoms of infestation and/or
enhancing one or more characteristics of growth
and/or yield in the plant, as compared to an untreated control plant.
Compositions of the present disclosure may be used to prevent, treat,
suppress, eliminate and/or reduce the
severity of infestations/infections of/by myriad phytopathogenic nematodes,
including, but not limited to, root-knot
nematodes, cyst nematodes, burrowing nematodes, root lesion nematodes, wilt
nematodes and rcniform nematodes. In
some embodiments, enzymes of the present disclosure are used to prevent,
treat, suppress, eliminate and/or reduce the
severity of an infestation/infection of a plant or plant part by one or more
Meloidogyne (e.g., M. arenaria, M.
graminicola, M. hap/a, Al. incognita, Al. javanica), Globodera (e.g., G.
pallida, G. rostochiensis), Heterodera (e.g., H.
avenae, H. filipjevi, H. glycines, H. schachtii), Pratylenchus (e.g., P.
penetrans, P. thornei, P. neglectus, P. zae, P.
vulnus, P. coffeae), Radopholus (e.g., R. similis), Ditylenchus (e.g., D.
dipsaci, D. angustus, D. desctructor, D. africanus,
D. myceliophagu.s, D. gigas), Bursaphelenchus (e.g., B. xylophilu.s, B.
mucronalus),Rotylenchus (e.g., R. reniformis, R.
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parvus), Xiphineina (e.g., X. index, X. italiae, X. viuttenezi, N.
diversicaudatun), and/or Nacobbus (e.g., N. aberrans).
Additional examples of nematodes that may be targeted by enzymes,
formulations, polynucleotides and organisms of the
present disclosure may be found in CAPINERA, HANDBOOK OF VEGETABLE PESTS
(2001) and Niblack, Nematodes, in
ILLINOIS AGRONOMY HANDBOOK (2008)
Compositions of the present disclosure may be used to prevent and/or treat
oomycete infestations/infections by,
for example, inhibiting adhesion of an oomycete to a surface, inhibiting entry
of an oomycete into a material, inhibiting
habitation by an oomycete, inhibiting the growth of an oomycete, inhibiting
the reproduction and/or proliferation of an
oomycete, degrading on one or more structural components of an oomycete (e.g.
cell wall components, such as
celluloses and other beta-glucans), killing an oomycete, and/or reducing one
or more symptoms of an oomycete
infestation/infection. In some embodiments, such inhibition is complete or
substantially complete, such that the
oomycete fails to inhabit/feed/grow/reproduce/proliferate at a rate effective
to initiate and/or sustain an appreciable
infestation/infection. For example, spraying a plant with an enzyme of the
present disclosure may inhibit an oomycete's
ability to adhere to the surface of a plant, inhibit an oomycete's ability to
enter into the plant, inhibit an oomycete's ability
to inhabit the plant, inhibit growth of an oomycete on/in the plant, inhibit
the reproduction and/or proliferation of an
oomycete on/in the plant, degrade one or more structural components of an
oomycete (e.g. one or more beta-glucans)
on/in the plant, and/or kill an oomycete, thereby reducing one or more
symptoms of infestation and/or enhancing one or
more characteristics of growth and/or yield in the plant, as compared to an
untreated control plant.
Compositions of the present disclosure may be used to prevent and/or treat
infestations/infections of myriad
phytopathogenic oomycetes, including, but not limited to, phytopathogenic
Albuginaceae, Peronosporaceae and
Pythiaceae, such as blights, mildews, molds, root rots and rusts In some
embodiments, proteins of the present disclosure
are used to prevent, treat, suppress, eliminate and/or reduce the severity of
an infestation/infection of a plant or plant part
by one or more Achlya, Albugo (e.g., A. candida),Aphanomyces (e.g., A.
cochlioides, A. euteiches, A. invadans, A. iridis,
A. raphani), Bremia (e.g., B. lactucae), Hyaloperonospora (e.g., H.
arabidopsidis), Peronospora (e.g., P. belbahrii, P.
destructor, P. effusa, P. farinose, P. ftilva, P. lotorum, P. manshurica, P.
parasitica, P. potentillae, P. rubi, P. schachtii,
P. sparsa, P. tabacina, P. trifolii, P. viciae), Phytophthora (e.g., P.
agathidicia, P. boehmeriae, P. cactorzun, P.
cambivora, P. capsica, P. cinnamomi, P. citricola, P. citrophthora, P.
cryptogea, P. drechsleri, P. erythroseptica, P.
fragariae, P. heveae, P. infestans, P. kernoviae, P. lateralis, P. megakaryam
P. megasperma, P. nicotianae, P.
palmivora, P. parasitica, P. ramorum, P. sojae, P. syringae), Plasm opara
(e.g., P. halstedii, P. viticola),
Pseudeoperonospora (e.g., P. cubensis, P. humuli), Pseudosclerospora (e.g., P.
philippinesis, P. sacchari, P. sorghi),
Pythium (e.g., P. acanthicum, P. aphanidermatum, P. ari,slo,sporum, P.
arrhenomanes, P. but/en, P. chondricold, P.
citrinum, P. cucurbitacearum, P. debaryanum, P. delicense, P. emineosum, P.
graminicola, P. heterothallicum, P.
hypogynum, P. insidiosum, P. irregulare, P. iwayamae, P. middletonii, P.
myriotylum, P. okanoganense, P. oopapillum,
P. paddicum, P. paroecandrum, P. perniciosum, P. porphyrae, P. rostratum, P.
scleroteichum, P. spinosum, P.
splendens, P. sulcatum, P. tardicrescens, P. tracheiphihtm, P. ultimum, P.
viohie, P. volutum),Saprolegnia (e.g., S.
parasitica), Sclerophthora (e.g., S. macrospora, S. rayssiae) and/or
Sclerospora (e.g., S. graminicola). Additional
examples of fungi that may be targeted using proteins, formulations,
polynucleotides and organisms of the present
disclosure may be found in Bradley, Managing Diseases, in ILLINOIS AGRONOMY
HANDBOOK (2008).
Compositions of the present disclosure may be particularly useful for
preventing, treating, suppressing,
eliminating and/or reducing the severity of infestations/infections of/by
phytopathogenic fungi and oomycetes, such as
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Albugo (e.g., A. candida, A. occidentalis), Alternaria (e.g. A. alternata, A.
alternantherae, A. arachidis, A. arborescens,
A. arbusti, A. blumeae, A. brassicae, A. brassicicola, A. burnsii. A.
carotiincultae, A. carthami, A. celosiae, A.
cinerariae, A. cari, A. conjuncta, A. cucumerina, A. dauci, A. dianthi, A.
dianthicola, A. eichhorniae, A. euphorblicola,
A. gaisen, A. helianthi, A. helianthi cola, A. hungarica, A. infectoria, A.
japonica, A. leucanthemi, A. linicola, A. longipes,
A. mall, A. molesta, A. padwickii, A. panax, A. perpunctulata, A. patroselini,
A. porn, A. quericola, A. radicina, A.
raphani, A. saponariae, A. selini, A. senecionis, A. smyrnii, A. solani, A.
sonchi, A. tenuissima, A. triticina, A. ventricosa,
A. zinniae), Blutneria (e.g., B. graminis), Botrytis (e.g., B. aclada, B.
allii, B. cinerea, B. elliptica, B. fabae, B.
squamosa), Ceratocystis (e.g., C. fimbriata), Colletotrichum, Diplodia (e.g.,
D. gossypina), Erwinia (e.g., E. atnylovora,
E. aphidi cola, E. carotovora, E. chrysanthemi, E. papayae, E. persicina, E.
psi dii, E. pyrifoliae, E. rhapontici, E.
tracheiphila), Fusarium (e.g., 1< graminearum, F. oxysporum,11. solani, F.
virgulifbrme), Geotrichum (e.g., G.
candidum), Gibberella (e.g., G. finikuroi, G. pulicaris, G. zeae), Gilbertella
(e.g., G. persicaria), Glomerella (e.g., G.
cingula la), Hyaloperono.spora (e.g., H. arabalop.sidis), Macrophomina (e.g.,
Xi pha.seolina), Magnaporthe (e.g., M.
grisea, Af. oryzae), Alelampsora (e.g., Al lini), Alonilinia (e.g., Al
jructicola), Alitcor (e.g., MI piriformis),
Mycosphaerella (e.g., Al. graminicola),Neolabraea (e.g., N. malicorticus),
Penicillium (e.g., P. digitatum, P. expansum,
P. italicum, P. rugulosum, P. verrucosum), Phakopsora (e.g., P. pachyrhizi),
Physalospora (e.g., P. obtusa),
Phytophthora (e.g., P. capsici, P. cinnamomi, P. infestans, P. parasitica, P.
ramorum, P. sojae), Plasmopara (e.g., P.
viticola), Pseudoperonospora (e.g., P. cubensis), Puccinia (e.g. , P.
asparagi, P. cacahata, P. graminis, P. kuehnii, P.
melanocephala, P. porn, P. punctiformis, P. recondita, P. schedonnardn, P.
sessihs, P. sorght, P. strnformis, P. tritici,
P. triticina), Pythium (e.g., P. butleri, P. ultimum), Rhizoctonia (e.g., R.
solani), Rhizopus (e.g., R. nigricans, R.
stolonijer), Sclerolinia (e.g., S. borealis, S. bulborum, S. homoeocarpa, S.
libertiana, S. minor, S. ricini, S. sclerotiorum,
S. sperm ophila, S. trifollorum),Septoria (e.g., S. cucurbitacearum, S.
glycines, S. lyco,spersici), Usti/ago (e.g., U.
esculenta, U maydis, U nuda), Zymoseptoria (e.g., Z. tritici).
Blast infestations/infections, such as those mediated by Alagnaporthe (e.g.,
Al. grisea, Al. oryzae), may be
prevented, treated, suppressed and/or eliminated with myriad compositions of
the present disclosure, including, but not
limited to, proteins exhibiting one or more activities belonging to EC 1.1.3
(e.g., 1.1.3.4, 1.1.3.25), 1.11.1 (e.g.,
1.11.1.6), 3.2.1 (e.g., 3.2.1.6) and/or 3.4.21 (and corresponding
formulations, polynucleotides and organisms). In some
embodiments, such infestations/infections may be prevented, treated,
suppressed and/or eliminated using one or more of
SEQ ID NOs: 1, 2, 5, 6, 10, 11, 14, 21, 99, 101, 116, 126, and enzymatically
active fragments/mutants/variants thereof.
In some embodiments, such infestations/infections may be prevented, treated,
suppressed and/or eliminated using a
combination of enzymes, such as one or more cellulases, one or more
hemicellulases and one or more xylanases; one or
more peptidases and one or more proteases; ; one or more cellulases, one or
more glucosidases and one or more
xylanases.
Blight infestations/infections, such as those mediated by Altemaria (e.g.. A.
alternata, A. carotiincultae, A.
pan ax, A. petroselini, A. solani, A. triticina), Colletotri chum, Fusarium
(e.g., F. graminearum), Gibberella (e.g., G.
zeae), Phytophthora (e.g., P. capsici , P. infestans, P. ramorum), may be
prevented, treated, suppressed and/or eliminated
with myriad compositions of the present disclosure, including, but not limited
to, proteins exhibiting one or more
activities belonging to EC 1.1.3 (e.g., 1.1.3.4, 1.1.3.25), 1.10.3 (e.g.,
1.10.3.2), 1.11.1 (e.g., 1.11.1.6), 3.1.1 (e.g.,
3.1.1.5), 3.2.1 (e.g., 3.2.1.1, 3.2.1.3, 3.2.1.4, 3.2.1.5, 3.2.1.6, 3.2.1.8,
3.2.1.11, 3.2.1.15, 3.2.1.39, 3.2.1.41, 3.2.1.58,
3.2.1.75, 3.2.1.78, 3.2.1.111), 3.4.11 (e.g., 3.4.11.1), 3.4.21 (e.g.,
3.4.21.19, 3.4.21.62) and/or 3.4.24 (e.g., 3.4.24.28)
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(and corresponding formulations, polynucleotides and organisms). In some
embodiments, such infestations/infections
may be prevented, treated, suppressed and/or eliminated using one or more of
SEQ ID NOs: 1, 6, 9, 10, 11, 12, 13, 14,
15, 16, 17, 19, 20, 21, 22, 24, 25, 29, 33, 42, 43, 44, 45, 46, 48, 99, 100,
101, 105, 109, 110, 114, 116, 117, 125, 126,
132, 149, and enzymatically active fragments/mutants/variants thereof. In some
embodiments, such
infestations/infections may be prevented, treated, suppressed and/or
eliminated using a combination of enzymes, such as
two or more cellulases; two or more pectinases; two or more glucanases; two or
more peptidases; two or more
pectinases; one or more cellulases, one or more hemicellulases and one or more
xylanases; one or more glucanses and
one or more xylanases; one or more amylases, one or more glucanases and one or
more xylanases.
Blotch infestations/infections, such as those mediated by Alycosphaerella
(e.g., Al. gramtnicola) and
Zymoseptoria (e.g., Z. tritici) may be prevented, treated, suppressed and/or
eliminated with myriad compositions of the
present disclosure, including, but not limited to, proteins exhibiting one or
more activities belonging to EC 1.1.3 (e.g.,
1.1.3.4, 1.1.3.25), 1.10.3 (e.g., 1.10.3.2), 1.11.1 (e.g., 1.11.1.6), 3.1.1
(e.g., 3.1.1.5), 3.2.1 (e.g., 3.2.1.1, 3.2.1.3, 3.2.1.6,
3.2.1.8, 3.2.1.11, 3.2.1.15, 3.2.1.39, 3.2.1.41, 3.2.1.58, 3.2.1.75, 3.2.1.78,
3.2.1.101, 3.2.1.109, 3.2.1.110, 3.2.1.112),
3.4.11 (e.g., 3.4.11.1), 3.4.21 (e.g., 3.4.21.62) and/or 3.4.24 (3.4.24.28)
(and corresponding formulations,
polynucleotides and organisms). In some embodiments, such
infestations/infections may be prevented, treated,
suppressed and/or eliminated using one or more of SEQ ID NOs: 1, 6, 9, 10, 11,
13, 14, 15, 17, 21, 22, 24, 25, 29, 42, 44,
45, 48, 101, 103, 114, 149, and enzymatically active
fragments/mutants/variants thereof. In some embodiments, such
infestations/infections may be prevented, treated, suppressed and/or
eliminated using a combination of enzymes, such as
one or more cellulases, one or more lyases and one or more pectinases; as one
or more cellulases, one or more
hemicellulases and one or more xylanases; two or more cellulases; two or more
glucanases; two or more pectinases; one
or more glucanses and one or more xylanases; one or more peptidases and one or
more proteases.
Downy mildew infestations/infections, such as those mediated by Bremia (e.g.,
B. lactucae), Hyaloperonospora
(e.g., H. arabidopsidis, H. parasitica), Peronospora (e.g., P. belbahrii, P.
destructor, P. elfitsa, P. .farinose, P. .fulva, P.
lotorum, P. manshurica, P. parasitica, P. potentillae, P. rubi, P. schachtii,
P. sparsa, P. tabacina. P. trifolii, P. viciae),
Peronosclerospora, Plasm opara (e.g., P. halstedit, P. viticola) and
Pseudoperonospora (e.g., P. cubensis, P. humult),
may be prevented, treated, suppressed and/or eliminated with myriad
compositions of the present disclosure, including,
but not limited to, proteins exhibiting one or more activities belonging to EC
3.2.1. (e.g., 3.2.1.8, 3.2.1.78) and/or 3.4.21
(e.g., 3.4.21.19, 3.4.21.62) (and corresponding formulations, polynucleotides
and organisms). In some embodiments,
such infestations/infections may be prevented, treated, suppressed and/or
eliminated using one or more of SEQ ID NOs:
22, 42, 43, 44, 45, and enzymatically active fragments/mutants /variants
thereof.
Povvdery, mildew infestations/infections, such as those mediated by Bhtmeria
(e.g., B. graminis), Erysiphe (e.g.,
E. cichoracearum, E. necator), Golovinomyces, Leveillula (e.g., L. taurica),
Alicrosphaera (e.g., Al. dttfusa), Oidium,
Phyllactinia, Podosphaera (e.g., P. aphanis, P. leucotricha, P. pannosa, P.
xanthii), Sphaerotheca and Uncinula, may be
prevented, treated, suppressed and/or eliminated with myriad compositions of
the present disclosure, including, but not
limited to, proteins exhibiting one or more activities belonging to EC 1.1.3
(e.g., 1.1.3.4), 3.2.1 (e.g., 3.2.1.8, 3.2.15)
and/or 3.4.21 (and corresponding formulations, polynucleotides and organisms).
In some embodiments, such
infestations/infections may be prevented, treated, suppressed and/or
eliminated using one or more of SEQ ID NOs: 1, 10,
22, 42, and enzymatically active fragments/mutants /variants thereof. In some
embodiments, such infestations/infections
may be prevented, treated, suppressed and/or eliminated using a combination of
enzymes, such as two or more
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pectinases.
Mold infestations/infections, such as those mediated by Botrytis (e.g., B.
cinerea, B. elliptica), Penicilhum (e.g.,
P. digitalum), Phylophthora (e.g., P. cap,sici, P. cinnamomi, P. ram orum, P.
sojae), may be prevented, treated,
suppressed and/or eliminated with myriad compositions of the present
disclosure, including, but not limited to, proteins
exhibiting one or more activities belonging to EC 1.1.3 (e.g., 1.1.3.4,
1.1.3.25), 1.10.3 (e.g., 1.10.3.2), 1.11.1 (e.g.,
1.11.1.6, 1.11.1.7), 3.2.1 (e.g., 3.2.1.1, 3.2.1.3, 3.2.1.4, 3.2.1.5, 3.2.1.6,
3.2.1.8, 3.2.1.11, 3.2.1.15, 3.2.1.17, 3.2.1.39,
3.2.1.41, 3.2.1.58, 3.2.1.75, 3.2.1.109, 3.2.1.110, 3.2.1.111, 3.2.1.112),
3.4.11 (e.g., 3.4.11.1), 3.4.21 (e.g., 3.4.21.62)
and/or 3.4.24 (e.g., 3.4.24.28) (and corresponding formulations,
polynucleotides and organisms). In some embodiments,
such infestations/infections may be prevented, treated, suppressed and/or
eliminated using one or more of SEQ ID NOs:
1, 6, 9, 10, 11, 12, 13, 14, 16, 17, 20, 21, 22, 24, 25, 29, 33, 41, 42, 44,
45, 48, 99, 101, 102, 111, 116, 119, 126, 129,
149, and enzymatically active fragments/mutants /variants thereof. In some
embodiments, such infestations/infections
may be prevented, treated, suppressed and/or eliminated using a combination of
enzymes, such as two or more
celluloses; two or more pectinases; one or more celluloses, one or more
hemicellulases and one or more xylanases; one or
more peptidases and one or more proteases; one or more amylases, one or more
glucanases and one or more
bacillolysins; ; one or more celluloses, one or more hemicellulases and one or
more xylanases; two or more peptidases;
one or more glucanses and one or more xylanases; one or more peptidases and
one or more proteases; one or more
celluloses, one or more glucosidases and one or more xylanases; one or more
celluloses, one or more furanosidases and
one or more xylanases.
Crown/fruit/root/stem rot infestations/infections, such as those mediated by
Colletotri chum, Fusarium (e.g., F.
solani, F. virguliforme), Phylophlhora (e.g., P. capsica, P. cinnamomi, P.
nicolianae, P. parasilica, P. sojae), Pythium
(e.g., P. graminicola, P. ul timum), Saprolegnia (e.g., S. parasitica), may be
prevented, treated, suppressed and/or
eliminated with myriad compositions of the present disclosure, including, but
not limited to, proteins exhibiting one or
more activities belonging to EC 1.1.3 (e.g., 1.1.3.4, 1.1.3.25), 1.11.1 (e.g.,
1.11.1.6), 3.2.1 (e.g., 3.2.1.6, 3.2.1.8, 3.2.1.11,
3.2.1.39, 3.2.1.58, 3.2.1.75, 3.2.1.109, 3.2.1.110, 3.2.1.111) and/or 3.4.21
(e.g., 3.4.21.19) (and corresponding
formulations, polynucleotides and organisms). In some embodiments, such
infestations/infections may be prevented,
treated, suppressed and/or eliminated using one or more of SEQ ID NOs: 1, 6,
9, 10, 11, 12, 13, 14, 15, 16, 17, 20, 21,
22, 24, 25, 29, 41, 42, 43, 44, 45, 48, and enzymatically active
fragments/mutants/variants thereof. In some
embodiments, such infestations/infections may be prevented, treated,
suppressed and/or eliminated using a combination
of enzymes, such as two or more celluloses; two or more pectinases; two or
more glucanases; two or more peptidases;
one or more amylases, one or more glucanases and one or more bacillolysins;
one or more celluloses, one or more lyases
and one or more pectinases; one or more celluloses, one or more hemicellulases
and one or more xylanases; one or more
glucanses and one or more xylanases; one or more peptidases and one or more
proteases.
Rust infestations/infections, such as thosc mediated by Albugo (e.g., A.
candida, A. occidentahs). Hemileia
(e.g., II. coffeicolu, II. vustutrix),Melumsporu (e.g., Phukopsoru (e.g.,
P. meibomiue, P. puchyrhizi), Pucciniu
(e.g., P. asparagi, P. cacahata, P. graminis, P. kuehnii, P. melanocephala, P.
porn, P. punctiformis, P. recondita, P.
schedonnardii, P. sessilis, P. song/it, P. striiformis, P. tritici, P.
triticina) and Urotnyces (e.g., U. appendiculatus), may
be prevented and/or treated with myriad compositions of the present
disclosure, including, but not limited to, proteins
exhibiting one or more activities belonging to EC 1.1.3 (e.g., 1.1.3.4),
1.10.3 (e.g., 1.10.3.2), 3.1.1 (e.g., 3.1.1.5), 3.2.1
(e.g., 3.2.1.1, 3.2.1.3, 3.2.1.4, 3.2.1.5, 3.2.1.6, 3.2.1.8, 3.2.1.15,
3.2.1.41, 3.2.1.58, 3.2.1.78), 3.4.11 (e.g., 3.4.11.1),
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3.4.21 (e.g., 3.4.21.19, 3.4.21.62) and/or 3.4.24 (e.g., 3.4.24.28) (and
corresponding formulations, polynucleotides and
organisms). In some embodiments, such infestations/infections may be
prevented, treated, suppressed and/or eliminated
using one or more of SEQ ID NOs: 1, 9, 13, 15, 16, 17, 19, 20, 23, 24, 29, 33,
42, 43, 44, 45, 46, 48, and enzymatically
active fragments/mutants /variants thereof. In some embodiments, such
infestations/infections may be prevented, treated,
suppressed and/or eliminated using a combination of enzymes, such as two or
more glucanases; one or more peptidases
and one or more proteases; and one or more amylases, one or more glucanases
and one or more bacillolysins
Wilt infestations/infections, such as those mediated by Fusarium (e.g., F.
oxysporum,), Phytophthora (e.g., P.
capsici, P. infestans, P. ramorum), may be prevented, treated, suppressed
and/or eliminated with myriad compositions of
the present disclosure, including, but not limited to, proteins exhibiting one
or more activities belonging to EC 1.1.3
(e.g., 1.1.3.4, 1.1.3.25), 1.10.3 (e.g., 1.10.3.2), 1.11.1 (e.g., 1.11.1.6),
3.1.1 (e.g., 3.1.1.5), 3.2.1 (e.g., 3.2.1.1, 3.2.1.3,
3.2.1.4, 3.2.1.5, 3.2.1.6, 3.2.1.8, 3.2.1.11, 3.2.1.15, 3.2.1.39, 3.2.1.41,
3.2.1.58, 3.2.1.75, 3.2.1.78, 3.2.1.111), 3.4.11
(e.g., 3.4.11.1), 3.4.21 (e.g., 3.4.21.19, 3.4.21.62) and/or 3.4.24 (e.g.,
3.4.24.28) (and coliesponding formulations,
polynucleotides and organisms). in some embodiments, such
infestations/infections may be prevented, treated,
suppressed and/or eliminated using one or more of SEQ ID NOs: 1, 6, 9, 10, 11,
12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 24,
25, 33, 42, 43, 44, 45, 46. 48, 99, 101, 114, 116, 125, 126, and enzymatically
active fragments/mutants/variants thereof.
In some embodiments, such infestations/infections may be prevented, treated,
suppressed and/or eliminated using a
combination of enzymes, such as two or more cellulases; two or more
pectinases; two or more glucanases; two or more
peptidases; two or more pectinases; one or more cellulases, one or more
hemicellulases and one or more xylanases; one
or more glucanses and one or more xylanases; one or more amylases, one or more
glucanases and one or more xylanases.
Proteins exhibiting activity belonging to EC 1.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusarium (e.g.,
F. graminearum, F. oxysporum, F. virgultforme), Alagnaporthe (e.g., Al. gri
sea, Al. oryzae), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamorni, P. infestans, P.
parasitica, P. ramorum, P. sojae) and
Zymoseptoria (e.g., Z. tritici). Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 1-8 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.1.3 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusarium (e.g.,
F. graminearum, F. oxysporum, F. virguliforme), Magnaporthe (e.g., M. grisea,
M. oryzae), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P. infestans, P.
paws itica, P. rumor um , P. softie) and
Zyinoseptoria (e.g., Z. tritici). Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 1-8 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
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Proteins exhibiting belonging to EC 1.1.3.4 (and corresponding formulations,
polynucleotides and organisms)
may be particularly useful for preventing, treating, suppressing, eliminating
and/or reducing the severity of
infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusarium (e.g.,
F. graminearum, F. oxysporum, F. virguliforme), Magnaporthe (e.g., M. grisea,
M. oryzae), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P. infestans, P.
parasitica, P. ramorum, P. so/ac) and
Zyinoseptoria (e.g., Z. tritici). Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 1-5 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.1.3.25 (now included in EC
1.1.99.18) (and corresponding
formulations, polynucleotides and organisms) may be particularly useful for
preventing, treating, suppressing,
eliminating and/or reducing the severity of infestations/infections of a plant
or plant part by Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguhforme), Magnaporthe
(e.g., AL grisea, AL oryzae),
Phytophthora (e.g., P. capsici, P. cinnamomi, P. ihfestans, P. parasitica, P.
ramorum, P. sojae) and Zymoseptoria (e.g.,
Z. tritici). Proteins comprising, consisting essentially of or consisting of
an amino acid sequence that is about/at least GO,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79,
80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs:
6-8 (and corresponding formulations, polynucleotides and organisms) may be
especially useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 1.10 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearurn, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici).
Proteins comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 9 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 1.10.3 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating anchor
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearurn, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici).
Proteins comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 9 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 1.10.3.2 (and
corresponding formulations,
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polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
gram inearum, F oxysporum, F virgulybrme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. trarci).
Proteins comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 9 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to EC 1.11 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxy.sporum, F virguliforme), Magnaporthe (e.g., XI grisea, 11/1. oryzae),
Phylophlhora (e.g., P. capsici, P cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria (e.g., 7
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94. 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 10-12 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.11.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botryas (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxysporum, F. virguhforme), Magnaporthe (e.g., M. grisea, lvi oryzae),
Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 10-12 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.11.1.6 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxysporum, F. virguhforme), Alagnaporthe (e.g., Al. grisea, M oryzae),
Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorurn, P. sojae) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 10-11 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.11.1.7 (and corresponding
formulations, polynucleotides and
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organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea). Proteins comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 12 (and
corresponding formulations, polynucleotides
and organisms) may be especially useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
such infestations/infections.
Proteins exhibiting activity belonging to EC 1.14 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., E graminearum,
oxysporutn, F. virguliforme), Magnaporthe (e.g., M. grisea, M oryzae),
Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasi lien, P. ramorum, P. .sojae) and Zymoseptoria (e.g., 7 In
tici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 99 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.14.99 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., E graminearum, E
oxysporum, E virguliforme), Magnaporthe (e.g., M. grisea, !vi oryzae),
Phytophihora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 99 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 1.14.99.56 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botryas (e.g., B.
cinerea), Fusarium (e.g., E graminearum,
oxysporum, F virguliforme), Magnaporthe (e.g., M. grisea, M oryzae),
Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. so/ac) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 99 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
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infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Eusarium (e.g., E graminearum, E
oxysporum, E virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 13 and 100-106 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to EC 3.1.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Eusarium (e.g., E graminearum,
oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 13, 100-103 and 105 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating.
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to EC 3.1.1.3 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., E graminearum, E
oxysporum, E virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 100-103 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to EC 3.1.1.5 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., E graminearum, E
oxysporum, E virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. fritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 13 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity' of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.1.1.11 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum,
oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
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69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 105 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusariutn (e.g.,
F. graminearum, F. oxysporum, F. virguliforme),11/lagnaporthe (e.g., M.
grisea, M. oryzae), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P. infestans, P.
parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. tritici).
Proteins comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ TD NOs: 14-41 and 109-
143 (and corresponding formulations,
polynucleotides and organisms) may be especially useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Blumerta (e.g., B.
gramints), Botrytis (e.g., B. cinerea), Fusartum (e.g.,
F. graminearum, F. oxysporum, F. virgulijOrme),Magnaporthe (e.g., M grisea, M.
oryzae), Phakopsora (e.g., P.
pachyrhizi), Phylophlhora (e.g., P. capsici, P. cinnamomi, P. infeslans, P.
parasilica, P. ramorum, P. sojae),
Pseudoperono,spora (e.g., P. cuben,sis) and Zymo,septoria (e.g., Z. triad).
Proteins comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 14-41 and 109-
143 (and corresponding formulations,
polynucleotides and organisms) may be especially useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.1 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxy,sporum, F virgulijOrme), Phakop,sora (e.g., P. pachyrhizi), Phytophthora
(e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 14-16 and 109-111 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.3 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytts (e.g., B.
cinerea), Fusarium (e.g., F. graminearum,
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oxysporum, E virguliforme), Phakopsora (e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 17-18 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.4 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxysporum, F virgulifbrme) and Phakopsora (e.g., P. pachyrhizi). Proteins
comprising, consisting essentially of or
consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ TD NOs: 19 and 114-
116 (and corresponding formulations,
polynucleotides and organisms) may be especially useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.6 (and corresponding
fommlations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxysporum, F. virgultfOrme), Magnaporthe (e.g., M. grisea, M oryzae),
Phakopsora (e.g., P. pachyrhizi) and
Zymoseptoria (e.g., Z. trilici). Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 20 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.7 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea) and Fusarium (e.g., F. graminearum, F.
oxysporum, F. virgulifOrme). Proteins comprising, consisting essentially of or
consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 117 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to EC 3.2.1.8 (and corresponding
formulations, polynucleotides and
organisms) may be particularly useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusarium (e.g.,
F. graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P.
cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising, consisting essentially of or
consisting of an amino acid sequence that is about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,
78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
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91, 92, 93, 94, 95, 96, 97. 98, 99 or 100 % identical to one or more of the
amino acid sequences set forth herein as SEQ
ID NOs: 21-24 and 119 (and corresponding formulations, polynucleotides and
organisms) may be especially useful for
preventing, treating, suppressing, eliminating and/or reducing the severity of
such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.11 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusariutn (e.g., F.
graminearum, F. ox-ysporum, F. virguliforme) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 25 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.15 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Blumeria (e.g., B. graminis), Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F. oxysporum, F. virgidifartne),
Phakopsora (e.g., P. pachyrhizi) and
Zymoseptoria (e.g., Z. tritici). Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94. 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 125 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.17 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 26 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.21 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Alagnaporthe (e.g., Al. gri sea, Al. oryzae),
Penicillium (e.g., P. digitatum, P. expansum, P. italicum, P. rugulosum, P.
verrucosum), and Phytophthora (e.g., P.
cupsici, P. cinnamomi, P. infestuns, P. parasitica, P. rumorum, P. softie).
Proteins comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 126 (and
corresponding formulations,
polynucleotides and organisms) may be especially useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of such infestations/infections.
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Proteins exhibiting activity belonging to belonging to EC 3.2.1.39 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botryns (e.g., B. cinerea), Fu,sarium (e.g.,
graminearum, F. oxysporum, F. virguliforme) and Zymoseptoria (e.g., Z.
tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 27-28 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.41 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botryns (e.g., B. cinerea), Fu.sarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici).
Proteins comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 29
(and corresponding formulations, polynucleotides and organisms) may be
especially useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.55 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Penicilhum (e.g., P. dig//alum, P. expan,s-um, P.
italicum, P. rugulosum, P. verrucosum). Proteins comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 129 (and corresponding formulations,
polynucleotides and organisms) may
be especially useful for preventing, treating, suppressing, eliminating and/or
reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.58 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botryns (e.g., B. cinerea), Fu,sarium (e.g.,
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. &Mei).
Proteins exhibiting activity belonging to belonging to EC 3.2.1.59 (and
corresponding formulations,
polynucleotides and organisms) may bc particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Zymoseptoria (e.g., Z. tritici). Proteins
comprising, consisting essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,
85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % identical to one or more of the amino acid sequences set forth
herein as SEQ ID NOs: 30 (and
corresponding formulations, polynucleotides and organisms) may be especially
useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
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Proteins exhibiting activity belonging to belonging to EC 3.2.1.73 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by 1-
qtsarium (e.g., F. gram inearum, P1 oxysporum,
F. virguliforme). Proteins comprising, consisting essentially of or consisting
of an amino acid sequence that is about/at
least 60, --- 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the
amino acid sequences set forth herein as SEQ
ID NOs: 132 (and corresponding formulations, polynucleotides and organisms)
may be especially useful for preventing,
treating, suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.75 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerect), Fusarium (e.g., F.
gram inearum, E oxysporum, F. virgultforme) and Zymoseptoria (e.g., 7 tri
lid). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 31-32 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.78 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxy,sporum, F. virgultforme), Pseudoperono,spora (e.g., P.
c:uben,si,$) and Zymo,septoria (e.g., Z. tritici).
Proteins comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 33
(and corresponding formulations, polynucleotides and organisms) may be
especially useful for preventing, treating,
suppressing, eliminating and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.101 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Fusarium (e.g., F. graminearum, F. oxysporum,
virgultlbrme) and Zymoseptoria (e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences sct forth hcrcin as SEQ ID NOs: 34 (and corresponding
formulations, polynucleotides and organisms)
may be especially useful for preventing, treating, suppressing, eliminating
and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.109 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerect), Fusariutn (e.g., F.
gram inearum, E orysporum, F. virgultforme) and Zymasepturia (e.g., Z. tri
lid). Proteins comprising, consisting
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essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 35-40 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.2.1.133 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusariutn (e.g., F.
graminearum, F. oxysporum, F. virguhforme) and Zymoseptoria (e.g., Z.
tritict). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 41 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4 (and
corresponding formulations, polynucleotides
and organisms) may be particularly useful for preventing, treating,
suppressing, eliminating and/or reducing the severity
of infestations/infections of a plant or plant part by Blumeria (e.g., B.
graminis), Botrytis (e.g., B. cinerea), Fusarium
(e.g., F. gramtnearum, F. oxysporum, F. virguhforme), Magnaporthe (e.g., M.
grtsea, M. oryzae), Phakopsora (e.g., P.
pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P. infestans, P.
parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. trilici).
Proteins comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 42-48 (and
corresponding formulations,
polynucleotides and organisms) may be especially useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4.11 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. fritici).
Proteins exhibiting activity belonging to belonging to EC 3.4.11.1 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici).
Proteins exhibiting activity belonging to belonging to EC 3.4.21 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Blumeria (e.g., B. graminis), Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F. oxysporum, F. virguhforme),
Magnaporthe (e.g., Al grisea, M. oryzae),
Phakopsora (e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P.
cinnamotni, P. infestans, P. parasitica, P. mmorutn,
P. so/an), P.seudoperono.spora (e.g., P. cub en.si.$) and Zymoseptoria (e.g.,
Z. triad). Proteins comprising, consisting
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essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 42-47 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4.21.19 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Fusarium (e.g., F. graminearuin, F. oxysporum,
F. virguliforme), Phakopsora (e.g., P. pachyrhizi), Phytophthora (e.g., P.
capsici, P. cinnamomi, P. infestans, P.
parasitica, P. ramorum, P. sojae) and Pseudoperonospora (e.g., P. cub ensis).
Proteins comprising, consisting essentially
of or consisting of an amino acid sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ TD NOs: 43 (and
corresponding formulations, polynucleotides
and organisms) may be especially useful for preventing, treating, suppressing,
eliminating and/or reducing the severity of
such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4.21.62 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgulijOrme), Phakopsora (e.g., P. pachyrhizi),
Phytophthora (e.g., P. capsici, P.
cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cub ensis) and Zymoseptoria
(e.g., Z. triad). Proteins comprising, consisting essentially of or consisting
of an amino acid sequence that is about/at
least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,
78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the
amino acid sequences set forth herein as SEQ
ID NOs: 44-47 (and corresponding formulations, polynucleotides and organisms)
may be especially useful for
preventing, treating, suppressing, eliminating and/or reducing the severity of
such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4.24 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi),
Phytophthora (e.g., P. capsici, P.
cinnamomi, P. infe,stans, P. para,sitica, P. ramorum, P. ,sojae) and
Zymoseptoria (e.g., Z. triad). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 48 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 3.4.24.28 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
gram inearum , F. oxysporum, F. virguliforme), Phakopsora (e.g., P.
pachyrhizi), Phyloph thorn (e.g., P. capsici, P.
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cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria
(e.g., Z. tritici). Proteins comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 48 (and corresponding
formulations, polynucleotides and organisms) may be especially useful for
preventing, treating, suppressing, eliminating
and/or reducing the severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 4.2 (and
corresponding formulations, polynucleotides
and organisms) may be particularly useful for preventing, treating,
suppressing, eliminating and/or reducing the severity
of infestations/infections of a plant or plant part by Botrytis (e.g., B.
cinerea), Fusarium (e.g., F. graminearum, F.
oxysporum, F virguldbrme), Penicillium (e.g., P. digitatum, P. expansum, P.
italicum, P. rugulosum, P. verrucosum),
and Zymoseptoria (e.g., Z. tritici). Proteins comprising, consisting
essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 149 (and corresponding formulations, polynucleotides and
organisms) may be especially useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of such infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 4.2.2 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgulifOrme), Penicillium (e.g., P. digitatum,
P. expansum, P. italicum, P. rugulosum,
P. verrucosum), and Zymoseptoria (e.g., Z. Iritici). Proteins comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID NOs: 149 (and corresponding
formulations, polynucleotides and organisms)
may be especially useful for preventing, treating, suppressing, eliminating
and/or reducing the severity of such
infestations/infections.
Proteins exhibiting activity belonging to belonging to EC 4.2.2.2 (and
corresponding formulations,
polynucleotides and organisms) may be particularly useful for preventing,
treating, suppressing, eliminating and/or
reducing the severity of infestations/infections of a plant or plant part by
Botrytis (e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Penicillium (e.g., P. digitatum,
P. expansum, P. italicum, P. rugulosum,
P. verruco,sum), and Zymoseptoria (e.g., Z. Irately Proteins comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID NOs: 149 (and corresponding
formulations, polynucleotides and organisms)
may be especially useful for preventing, treating, suppressing, eliminating
and/or reducing the severity of such
infestations/infections.
It is to be understood that compositions of the present disclosure needn't be
toxic to be effective. As noted
above, proteins of the present disclosure may exert their effects through
various non-lethal means, such as reducing the
attraction of a pest to a treated surface, inhibiting feeding, etc. Moreover,
proteins and compositions of the present
disclosure-even those capable of exerting a toxic effect-may be used in non-
lethal doses to enhance the efficacy of
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and/or expand the target pest range of various chemical pesticides and
biological pesticides.
The present disclosure extends to methods of using compositions of the present
disclosure (e.g., proteins,
formulations, polynucleotides and organisms of the present disclosure) for
preventing, treating, suppressing, eliminating
and/or reducing the severity of infestations/infections of/by horticultural
pests, such as phytopathogenic acarids, bacteria,
fungi, gastropod, insects, nematodes, oomycetes protozoa and weeds.
In some embodiments, a protein exhibiting activity belonging to EC 1.1 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. graminis), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguhforme), Alagnaporthe
(e.g., Al. grisea, Al. oryzae), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae) and
Zymoseptoria (e.g., Z. tritici). For example, an oxidoreductase, optionally a
protein comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100% identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 1-8, (or a
corresponding formulation, polynucleotide,
or organism) may be applied to a plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of
such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.1.3 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. graminis), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguliforme), Magnaporlhe
(e.g., M. gri sea, Al. oryzae), Phakopsora
(e.g., P. pachyrhizi), Phylophlhora (e.g., P. capsici, P. cinnamomi, P.
infe,stan,s, P. para,sitica, P. ramorum, P. ,so j ae) and
Zymoseptoria (e.g., Z. tritici). For example, a oxidoreductase (with oxygen as
an acceptor), optionally a protein
comprising, consisting essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,
85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % identical to one or more of the amino acid sequences set forth
herein as SEQ ID NOs: 1-8 (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting belonging to EC 1.1.3.4 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. gramini,$), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguliforme), Magnaporthe
(e.g., M. gri sea, Al. oryzae), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. so/ac) and
Zymoseptoria (e.g., Z. tritici). For example, a glucose oxidasc, optionally a
protcin comprising, consisting essentially of
or consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 1-5, (or a
corresponding formulation, polynucleotide,
or organism) may be applied to a plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of
such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.1.3.25
(now included in EC 1.1.99.18)
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(or a corresponding formulation, polynucleotide, or organism) is used to
prevent, treat, suppress, eliminate and/or reduce
the severity of an infestation/infection of a plant or plant part of/by one or
more Botrytis (e.g., B. cinerea), Fusarium
(e.g., F. gram inearum, 1,1 oxysporum , P. virgulybrme), Magnaporthe (e.g., M.
gri,sea, Al. oryzae), Phyloph hora (e.g., P.
capsici, P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and
Zymoseptoria (e.g., Z. tritici). For example,
a cellobiose dehydrogenase, optionally a protein comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 6-8, (or a corresponding
formulation, polynucleotide, or organism) may be
applied to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 1.10 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a oxidase, optionally a protein comprising, consisting essentially of
or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 9, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 1.10.3 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a oxidase, optionally a protein comprising, consisting essentially of
or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 9, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 1.10.3.2 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a laccasc, optionally a protcin comprising, consisting essentially of
or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 9, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.11 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
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infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., PI
graminearum, E oxysporum, F. virguhforme), Magnaporthe (e.g., M. grisea, M.
oryzae), Phytophthora (e.g., P. capsici,
P. cinnamomi, P. infrstan,s, P. parasitica, P. ramorum, P. so/ac) and
Zymoseptoria (e.g., Z. traici). For example, a
peroxidase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 10-12, (or a corresponding formulation, polynucleotide,
or organism) may be applied to a plant
or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.11.1 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum, F oxysporum, F. virguhjorme), Magnaporthe (e.g., M. gri,sea, M.
oryzae), Phyloph thorn (e.g., P. capsici,
P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and
Zyrnoseptoria (e.g., Z. tritici). For example, a
peroxidase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86. 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 10-12, (or a corresponding formulation, polynucleotide,
or organism) may be applied to a plant
or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.11.1.6
(or a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgzdiforme), Magnaporthe (e.g., M. grisea, M.
oryzae), Phytophthora (e.g., P. capsici,
P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. so/ac) and
Zymoseptoria (e.g., Z. tritici). For example, a
catalase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 10-11, (or a corresponding formulation, polynucleotide,
or organism) may be applied to a plant
or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.11.1.7
(or a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea). For example, a peroxidase,
optionally a protein comprising, consisting essentially of or consisting of an
amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 12, (or a corresponding formulation, polynucleotide, or organism) may be
applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.14 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrylis
(e.g., B. cinerea), FLISOritIM (e.g., F.
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graminearum, F oxysporum, F virguliforme), Magnaporthe (e.g., Al. grisea. M.
oryzae), Phytophthora (e.g., P. capsici,
P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and
Zymoseptoria (e.g., Z. tritici). For example, a
oxygenase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 99, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.14.99 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), liusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Magnaporthe (e.g., M. gri sea, Al.
oryzae), Phytophthora (e.g., P. capsici,
cinnamomi , P infestans, P parasitica, P. ramorum, F softie) and Zymoseptori a
(e.g., 7 tri lici). For example, a
monooxygenase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that
is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94. 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 99, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 1.14.99.56
(or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Bolt:vas
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum , F. oxy.sporum, F. virgultfiffme), Magnaporthe (e.g., Al gri,sea,
Al. oryzae), Phytophthora (e.g., P. cap,sici,
P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and
Zymoseptoria (e.g., Z. tritici). For example, a lytic
cellulose monooxygenase, optionally a protein comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 99, (or a corresponding formulation,
polynucleotide, or organism) may be
applied to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.1 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Bottyas
(e.g., B. cinerea), Fusartum (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a hydrolasc, optionally a protein comprising, consisting essentially
of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, '76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 13, and 100-106 (or a corresponding formulation,
polynucleotide, or organism) may be applied
to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.1.1 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
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infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., PI
graminearum, E oxysporum, F. virgultforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a carboxylic ester hydrolase, optionally a protein comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID NOs: 13, 100-103 and 105, (or a
corresponding formulation, polynucleotide,
or organism) may be applied to a plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of
such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.1.1.3 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum , F oxysporum, F. virguhlorme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseporia (e.g., Z. trilici). For
example, a triacylglycerol lipase, optionally a protein comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 100-103, (or a corresponding
formulation, polynucleotide, or organism) may
be applied to a plant or plant part to prevent, treat, suppress, eliminate
and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.1.1.5 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a lysophospholipase, optionally a protein comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 13, (or a corresponding formulation,
polynucleotide, or organism) may be
applied to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.1.1.11
(or a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a pectinesterase, optionally a protein comprising, consisting
essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,
71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83,
84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%
identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 105, (or a corresponding
formulation, polynucleotide, or organism) may be
applied to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2 (or a
corresponding formulation,
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polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. graminis),Botrytis (e.g., B. cinerea),
Fusarium (e.g., PI gram inearum , P1 oxysporum , P1 virgulybrme), Magnaporihe
(e.g., M. gri,sea , M. oryzae), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. tritici). For
example, a glycosylase, optionally a
protein comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 14-41 and
109-143, (or a corresponding formulation, polynucleotide, or organism) may be
applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2.1 (or a
corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. grarninis), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguliforme), Magnaporthe
(e.g., AL grisea, AL oryzae), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P. CinnaMOMi, P.
infestans, P. parasitica. P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. tritici). For
example, a glyeosidase, optionally a
protein comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 'A identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 14-41 and
109-143, (or a corresponding formulation, polynucleotide, or organism) may be
applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2.1.1 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi),
Phytophthora (e.g., P. capsici, P.
cinnamomi, P. infestans, P. parasinca, P. ramorum, P. sojae) and Zymoseptoria
(e.g., 7 tritici). For example, an alpha-
amylase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86. 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 14-16 and 109-111, (or a corresponding formulation,
polynucleotide, or organism) may be
applied to a plant or plant part to prevent, treat, suppress, eliminate and/or
reducing the severity of such
infestations/infections.
In somc embodiments, a protein exhibiting activity belonging to EC 3.2.1.3 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a glucan 1,4-alpha-glucosidase, optionally a protein comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
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acid sequences set forth herein as SEQ ID NOs: 17-18, (or a corresponding
formulation, polynucleotide, or organism)
may be applied to a plant or plant part to prevent, treat, suppress, eliminate
and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2.1.4 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. ox-ysporum, F. virguhforme) and Phakopsora (e.g., P.
pachyrhizi). For example, a cellulase, optionally
a protein comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62,
63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or more of the amino acid
sequences set forth herein as SEQ ID NOs: 19 and
114-116, (or a corresponding formulation, polynucleotide, or organism) may be
applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
in some embodiments, a protein exhibiting activity belonging to EC 3.2.1.6 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgulilbrme),Magnaporthe (e.g., Al. gri sea, M.
oryzae), Phakopsora (e.g., P.
pachyrhizi) and Zymoseptoria (e.g., Z. tritici). For example, an endo-1,3(4)-
beta-glucanase, optionally a protein
comprising, consisting essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72. 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,
85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % identical to one or more of the amino acid sequences set forth
herein as SEQ ID NOs: 20, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2.1.7 (or
a corresponding formulation,
polynucleotide, or organism) is used to prevent, treat, suppress, eliminate
and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea) and Fusarium (e.g., F.
gram inearum, F oxysporum, F virguhforme). For example, a inulinase,
optionally a protein comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78. 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 117, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to EC 3.2.1.8 (or
a corresponding formulation,
polynucleotide, or organism) is uscd to prevent, treat, supprcss, eliminate
and/or reduce thc severity of an
infestation/infection of a plant or plant part of/by one or more Blumeriu
(e.g., B. graminis), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporutn, F. virguliforme), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g.,
P. capsici, P. cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and
Zymoseptoria (e.g., Z. tritici). For example, a endo-1,4-beta-xylanase,
optionally a protein comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
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identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 21-24 and 119, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.11 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. ox-ysporum, F. virguliforme) and Zymoseptoria (e.g., Z.
tritici). For example, a dextranase, optionally a
protein comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 25, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
in some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.15 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g.. B. grarninis), Bottytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virgulilbrtne), Phakopsora
(e.g., P. pachyrhizi) and Zymoseptoria
(e.g., Z. tritici). For example, a endo-polygalacturonase (pectinase),
optionally a protein comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78. 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 'A
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 125, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.17 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea). For example, a lysozyme,
optionally a protein comprising, consisting essentially of or consisting of an
amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98. 99 or 100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 26, (or a corresponding formulation, polynucleotide, or organism) may be
applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.21 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Magnaporthe
(e.g., M. grisea, Al. oryzae), Penicillium
(e.g., P. digitutum, P. exptinsutn, P. Willem, P. rugulosum, P. verrueosum),
and Phytophthoru (e.g., P. eapsiej, P.
cinnamomi, P. injestans, P. parasitica, P. rainorutn, P. sojae). For example,
a beta-glucosidase, optionally a protein
comprising, consisting essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64,65,
66, 67, 68, 69, 70, 71, 72. 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,
85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % identical to one or more of the amino acid sequences set forth
herein as SEQ ID NOs: 126, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
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suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.39 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusartum (e.g., F.
graminearum, F. oxysporum, F. virgultforme) and Zymoseptoria (e.g., Z.
tritici). For example, a glucan endo-1,3-beta-D-
glucosidase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 27-28, (or a corresponding formulation, polynucleotide,
or organism) may be applied to a plant
or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.41 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Bottytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, a pullulanasc, optionally a protein comprising, consisting
essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 29, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.55 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Penicillium
(e.g., P. digitatum, P. expansum, P. italicum,
P. rugulosum, P. verrucosum). For example, an alpha-L-arabinofuranosidase,
optionally a protein comprising, consisting
essentially of or consisting of an amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
identical to one or more of the amino acid sequences set forth herein as SEQ
ID NOs: 129, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.58 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Bottytis
(e.g., B. cinerea), Fusartum (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritich. For
example, a glucan 1,3-beta-glucosidase (or a corresponding formulation,
polynucleotide, or organism) may bc applied to
a plant or plant part to prevent, treat, suppress, eliminate and/or reducing
the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.59 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Zymoseptoria
(e.g., Z. tritiei). For example, a glucan endo-
1,3-alpha-glucosidase, optionally a protein comprising, consisting essentially
of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
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87, 88, 89, 90, 91, 92, 93. 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 30, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.73 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Fusarium
(e.g., F. graminearum, F. oxysporum, F.
virguliforme). For example, a licheninase, optionally a pmtein comprising,
consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID NOs: 132, (or a corresponding
formulation, polynucleotide, or organism) may
be applied to a plant or plant part to prevent, treat, suppress, eliminate
and/or reducing the severity of such
infestations/infections.
in some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.75 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgulilbrme) and Zymoseptoria (e.g., Z.
tritici). For example, a glucan endo-1,6-beta-
glucosidase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 31-32, (or a corresponding formulation, polynucleotide,
or organism) may be applied to a plant
or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.78 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Pseudoperonospora (e.g., P.
cubensis) and Zymoseptoria (e.g., Z. tritiO.
For example, a mannan endo-1,4-beta-mannosidase, optionally a protein
comprising, consisting essentially of or
consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 33, (or a
corresponding formulation, polynucleotide,
or organism) may be applied to a plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of
such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.101 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Fusarium
(e.g., F. graminearum, F. oxysporum, F.
virguliforme) and Zymoseptoria (e.g., Z. tritici). Pro For example, a protein
teins comprising, consisting essentially of or
consisting of an amino acid sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,
95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth herein as SEQ ID NOs: 34 (or a
corresponding formulation, polynucleotide,
or organism) may be applied to a plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of
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such infestations/infections. For example, a mannan endo-1,6-alpha-mannosidase
(or a corresponding formulation,
polynucleotide, or organism) may be applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing
the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.109 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgtdiforme) and Zymoseptoria (e.g., Z.
tritici). For example, a endogalactosaminidase,
optionally a protein comprising, consisting essentially of or consisting of an
amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100% identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 35-40, (or a corresponding formulation, polynucleotide, or organism) may
be applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
in some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.2.1.133 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgulilbrme) and Zymoseptoria (e.g., Z.
tritici). For example, a glucan 1,4-alpha-
maltohydrase, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,
76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 41, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. gran-lints), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguliforme), Magnaporthe
(e.g., M. grisea, M. oryzae), Phak-opsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. cap sici, P. cinnamomi, P.
infestans, P parasitica, P ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. tritici). For
example, a peptidases, optionally a protein
comprising, consisting essentially of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65,
66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84,
85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % identical to one or more of the amino acid sequences set forth
herein as SEQ ID NOs: 42-48, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.11 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi)
and Zymoseptoria (e.g., Z. tritici). For
example, an aminopeptidase (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.11.1 (or a corresponding
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formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum, F oxy,sporum, F. virgultibrme), Phakop,sora (e.g., P.
pachyrhizi) and Zymo,septoria (e.g., Z. triad). For
example, a leucyl aminopeptidase (or a corresponding formulation,
polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat, suppress, eliminate and/or reducing the
severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.21 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Blumeria
(e.g., B. graminis), Botrytis (e.g., B. cinerea),
Fusarium (e.g., F. graminearum, F. oxysporum, F. virguhforme), Alagnaporthe
(e.g., Al. grisea, Al. oryzae), Phakopsora
(e.g., P. pachyrhizi), Phytophthora (e.g., P. capsici, P. cinnamomi, P.
infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria (e.g., Z. tritici). For
example, a serine endopeptidase,
optionally a protein comprising, consisting essentially of or consisting of an
amino acid sequence that is about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,
79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,
92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID
NOs: 42-47, (or a corresponding formulation, polynucleotide, or organism) may
be applied to a plant or plant part to
prevent, treat, suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.21.19 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Fusarium
(e.g., F. graminearum, F. oxysporum, F.
virgtdiforme), Phakopsora (e.g., P. pachyrhizi), Phylophlhora (e.g., P.
capsici, P. cinnamomi, P. infestans, P. parasitica,
P. ramorum, P. ,sojae) and Ps-eudoperono,spora (e.g., P. cubensis). For
example, a glutamyl endopeptidase, optionally a
protein comprising, consisting essentially of or consisting of an amino acid
sequence that is about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,
96, 97, 98, 99 or 100 % identical to one or more of the amino acid sequences
set forth herein as SEQ ID NOs: 43, (or a
corresponding formulation, polynucleotide, or organism) may be applied to a
plant or plant part to prevent, treat,
suppress, eliminate and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.21.62 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Bohytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum, F oxy,sporum, F. virgultibrme), Phakop,sora (e.g., P.
pachyrhizi), Phylophthora (e.g., P. capsici, P.
cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae),
Pseudoperonospora (e.g., P. cubensis) and Zymoseptoria
(e.g., Z. tritici). For example, a subtilisin, optionally a protein
comprising, consisting essentially of or consisting of an
amino acid sequence that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,
81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino
acid sequences set forth herein as SEQ ID NOs: 44-47, (or a corresponding
formulation, polynucleotide, or organism)
may be applied to a plant or plant part to prevent, treat, suppress, eliminate
and/or reducing the severity of such
infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.24 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
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infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., PI
graminearum, E oxysporum, F. virguhforme), Phakopsora (e.g., P. pachyrhizi),
Phytophthora (e.g., P. capsici, P.
cinnamomi, P. inje,stans, P. parasarca, P. ramorum, P. so/ac) and Zymoseptoria
(e.g., Z. triad). For example, a
metalloendopeptidase, optionally a protein comprising, consisting essentially
of or consisting of an amino acid sequence
that is about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one
or more of the amino acid sequences set forth
herein as SEQ ID NOs: 48, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 3.4.24.28 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum , F. oxysporum, F. virgulijorme), Phakop.sora (e.g., P.
pachyrhizi), Phyloph thorn (e.g., P. capsici , P.
cinnamomi, P. infestans, P. parasitica, P. ramorum, P. sojae) and Zymoseptoria
(e.g., Z. tritici). For example, a
bacillolycin, optionally a protein comprising, consisting essentially of or
consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, '70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86. 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 % identical to one or
more of the amino acid sequences set forth
herein as SEQ ID NOs: 48, (or a corresponding formulation, polynucleotide, or
organism) may be applied to a plant or
plant part to prevent, treat, suppress, eliminate and/or reducing the severity
of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 4.2 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virgzdiforme), Penicillium (e.g., P. digitatum,
P. expansum, P. itahcum, P. rugzdosum,
P. verrucosum), and Zymoseptoria (e.g., Z. tritici). For example, a carbon-
oxygen lyase, optionally a protein comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 149, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 4.2.2 (or a corresponding
formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
graminearum, F. oxysporum, F. virguliforme), Penicillium (e.g., P. digitatum,
P. expansum, P. italicum, P. rugulosum,
P. verrucosum), and Zymoseptoria (e.g., Z. tritici). For example, a
polysaccharide lyasc, optionally a protein comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 149, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
In some embodiments, a protein exhibiting activity belonging to belonging to
EC 4.2.2.2 (or a corresponding
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formulation, polynucleotide, or organism) is used to prevent, treat, suppress,
eliminate and/or reduce the severity of an
infestation/infection of a plant or plant part of/by one or more Botrytis
(e.g., B. cinerea), Fusarium (e.g., F.
gram inearum, F oxy.sporum, F. virguldbrme), Penici I hum (e.g., P. digitalum,
P. expansum, P. it alicum , P. rugulo,yum,
P. verrucosum), and Zymoseptoria (e.g., Z. tritici). For example, a pectate
lyase, optionally a protein comprising,
consisting essentially of or consisting of an amino acid sequence that is
about/at least 60, 61, 62, 63, 64, 65, 66, 67, 68,
69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87,
88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or
100 % identical to one or more of the amino acid sequences set forth herein as
SEQ ID NOs: 149, (or a corresponding
formulation, polynucleotide, or organism) may be applied to a plant or plant
part to prevent, treat, suppress, eliminate
and/or reducing the severity of such infestations/infections.
Compositions of the present disclosure may be applied to any plant type,
including, but not limited to, row
crops and vegetables. In some embodiments, compositions of the present
disclosure are formulated for the treatment of
one or more plants selected from the families Amaranthaceae (e.g., chard,
spinach, sugar beet, quinoa), Asteraceae (e.g.,
artichoke, asters, chamomile, chicory, chrysanthemums, dahlias, daisies,
echinacea, goldenrod, guayule, lettuce,
marigolds, safflower, sunflowers, zinnias), Brassicaceae (e.g., arugula,
broccoli, bok choy, Brussels sprouts, cabbage,
cauliflower, canola, collard greens, daikon, garden cress, horseradish, kale,
mustard, radish, rapeseed, rutabaga, turnip,
wasabi, watercress, Arabidopsis thaliana), Caricaceae (e.g., papaya),
Cucurbitaceae (e.g., cantaloupe, cucumber,
honeydew, melon, pumpkin, squash (e.g., acorn squash, butternut squash, summer
squash), watermelon, zucchini),
Fabaceae (e.g., alfalfa, beans, carob, clover, guar, lentils, mesquite, peas,
peanuts, soybeans, tamarind, tragacanth, vetch),
Malvaceae (e.g., cacao, cotton, durian, hibiscus, kenaf, kola, okra), Poaceae
(e.g., bamboo, barley, corn, fonio, lawn
grass (e.g., Bahia grass, Bennudagrass, bluegrass, Buffalograss, Centipede
grass, Fescue, or Zoysia), millet, oats,
ornamental grasses, rice, rye, sorghum, sugar cane, triticale, wheat and other
cereal crops, Polygonaceae (e.g.,
buckwheat), Rosaccac (e.g., almonds, apples, apricots, blackberry, blueberry,
cherries, peaches, plums, quinces,
raspberries, roses, strawberries), Solanaceae (e.g., bell peppers, chili
peppers, eggplant, petunia, potato, tobacco, tomato)
and Vitaccac (e.g., grape).
Non-limiting examples of plants that may be treated with compositions of the
present disclosure include plants
sold under the ACCELERON , AGRIPRO1-3, AGRISURE , AGROESTE , AGVENTURE ,
ALFOREXTM,
ASGROW , AQUAMAX , BOLLGARD JJTM, BOLLGARDTM 3, BREVANTTm, CHANELTM,
CONFIDORTM,
COR 'EVA AGRISCIENCETM. CORVUSTM. CREDENZ , CROPSTARTm, DAIRYLANDTM. DEKALB ,
DELTAPINETm, DERUIILRTM, DROUGHTGARD , ENLIST E3CD, ENOGENO, FIBERMAX ,
GAUCHOTM,
GENU1TY , GOLDENHARVEST , HOEGEMEYERTm, 1NTACTA RR2 PRO'TM, INVIGORV, LIBERTY
LINK ,
NEXGROW1?), NUTECH SEED , OPTIMUM , PHYTOGEN , PIONEER , QROME ,
RIB COMPLETE ,
ROUNDUP READY , ROUNDUP READY 2 YIELD , ROUNDUP READY 2 XTEND , SEMETES
AGROCERESTM, SEMINISTm, SMARTSTAXO, STONEVILLE , SYNGENTAO, TRUFLEXTm, VT
DOUBLE
PRO , VT TRIPLE PRO , YIELDGARD , YIELDGARD VT ROOTWORM/RR210, YIELDGARD VT
TRIPLE
and/or XTENDFLEXTm tradenames.
Compositions of the present disclosure may be applied to any part/portion of a
plant. In some embodiments, the
compositions are applied to plant propagation materials (e.g., cuttings,
rhizomes, seeds and tubers). In some
embodiments, the compositions are applied to the roots of a plant. In some
embodiments, the compositions are applied to
the foliage of a plant. In some embodiments, the compositions are applied to
both the roots and the foliage of a plant. In
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some embodiments, the compositions are applied to plant propagation materials
and to the plants that grow from said
plant propagation materials.
Compositions of the present disclosure may be applied to any plant growth
medium, including, but not limited
to, soil.
Compositions of the present disclosure may be applied to plants, plant parts
and/or plant growth media in any
suitable manner, including, but not limited to, on-seed application, in-furrow
application and foliar application.
Compositions of the present disclosure may be applied using any suitable
method(s), including, but not limited
to, coating, dripping, dusting, encapsulating, fogging, immersing, spraying,
and soaking. Batch systems, in which
predetermined batch sizes of material and composition are delivered into a
mixer, may be employed. Continuous
treatment systems, which are calibrated to apply composition at a predefined
rate in proportion to a continuous flow of
material, may also be employed. In some embodiments, compositions of the
present disclosure are applied using a boom
sprayer or an orchard sprayer.
in some embodiments, compositions of the present disclosure are applied
directly to plant propagation material
(e.g., seeds). According to some embodiments, plant propagation materials are
soaked in a composition of the present
disclosure for at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.25,
1.5, 1.75, 2, 3, 4, 5, 6, 9, 12, 15, 18, 21, 24, 36, 48
hours. According to some embodiments, plant propagation materials are coated
with the compositions. Plant propagation
materials may be coated with one or more additional layers (e.g., one or more
protective layers that serve to enhance the
stability and/or activity of an enzyme/organism of the present disclosure
and/or one or more sequestration layers
comprising substances that may reduce the stability and/or activity of an
enzyme/organism of the present disclosure if
included in the same layer as the enzyme/organism of the present disclosure).
In some embodiments, the coating
comprises, consists essentially of, or consists of a composition of the
present disclosure and a drying powder.
In some embodiments, compositions of the present disclosure are applied
directly to a plant growth medium
(e.g., a soil). According to some embodiments, the compositions are applied in
the vicinity of a plant propagation
material (e.g., a seed). According to some embodiments, the compositions arc
applied to the root zone of a plant
According to some embodiments, the compositions are applied using a drip
irrigation system.
In some embodiments, compositions of the present disclosure are applied
directly to plants. According to some
embodiments, the compositions are fogged, misted, sprayed and/or sprinkled
onto the plant(s) to be treated (e.g., foliar
sprays).
In some embodiments, compositions of the present disclosure are applied to
harvested plants and/or plant parts.
Individual components of the compositions (e.g., proteins of the present
disclosure and chemical pesticides)
may be separately or together. For example, in some embodiments, compositions
of the present disclosure may be
incorporated into integrated pest management strategies (e.g., a formulation
comprising one or more proteins of the
prcscnt disclosure may bc applied to an orchard/vincyard as part of an
integrated pest management strategy that includes
separate applications of 2, 3, 4, 5 or more distinct pesticides in a rotation
designed to reduce/prevent chemical pesticide-
induced phytotoxicity and/or pest resistance).
In some embodiments, compositions of the present disclosure are freeze- spray-
or spray-freeze-dried and then
applied to plants/plant parts. For examples, in some embodiments, a
formulation comprising and enzyme/organism of the
present disclosure and one or more stabilizing components (e.g., one or more
maltodextrins having a DEV of about 15 to
about 20) is freeze- spray- or spray-freeze-dried, mixed with a drying powder
(e.g., a drying powder comprising calcium
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stearate, attapulgite clay, montmorillonite clay, graphite. magnesium
stearate, silica (e.g., fumed silica, hydrophobically-
coated silica and/or precipitated silica) and/or talc), then coated on seed
that was been pre-treated with one or more
adhesives (e.g., an adhesive composition comprising one or more maltodextrins,
one or more mono-, di- or
oligosaccharides, one or more peptones, etc.), one or more pesticides and/or
one or more plant signal molecules (e.g.,
one or more LC0s).
Compositions of the present disclosure may be applied to plants, plant parts
and/or plant growth media in any
suitable amount(s)/concentration(s).
Compositions of the present disclosure may be used to prevent and/or treat
infestations/infections of/by
horticultural pests at any time, including, prior to planting, at the time of
planting, after planting, prior to germination,
after germination, prior to seedling emergence, at the time of seedling
emergence, after seedling emergence, prior to the
vegetative stage, during the vegetative stage, after the vegetative stage,
prior to the reproductive stage, during the
reproductive stage, after the reproductive stage, prior to flowering, at the
time of flowering, after flowering, prior to
fruiting, at the time of fruiting, after fruiting, prior to ripening, at the
time of ripening, after ripening, prior to harvest, at
the time of harvest, and after harvesting. Indeed, compositions of the present
disclosure may be used to extend the shelf-
life of harvested products by preventing, treating, suppressing, eliminating
and/or reducing the severity of
infestations/infections of/by acarids, bacteria, fungi, insects, oomycetes and
protozoa for many weeks/months post-
harvest.
Compositions of the present disclosure may be applied to plants, plant parts
and/or plant growth media at any
time, including, but not limited to, prior to planting, at the time of
planting, after planting, prior to germination, at the
time of germination, after germination, prior to seedling emergence, at the
time of seedling emergence, after seedling
emergence, prior to the vegetative stage, during the vegetative stage, after
the vegetative stage, prior to the reproductive
stage, during the reproductive stage, after the reproductive stage, prior to
flowering, at the time of flowering, after
flowering, prior to fruiting, at the time of fruiting, after fruiting, prior
to ripening, at the time of ripening, after ripening,
prior to harvest, at the time of harvest, and after harvesting.
In some embodiments, compositions of the present disclosure are applied to
plant propagation materials (e.g.,
seeds) about/at least 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 28, 32, 36, 40, 44, 48,
52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104 weeks prior to
planting.
In some embodiments, compositions of the present disclosure are applied to
plant propagation materials (e.g.,
seeds) at the time of planting.
In some embodiments, compositions of the present disclosure are applied to
plant propagation materials (e.g.,
seeds) after planting but before germination.
In some embodiments, compositions of the present disclosure are applied to
plants following emergence.
In some embodiments, compositions of the present disclosure arc applied to a
plant or plant part pre-harvest
(i.e., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more days
before the plant or plant part is (to be) harvested).
In some embodiments, compositions of the present disclosure are applied to a
plant or plant part post-harvest
(i.e., after the plant or plant part has been harvested).
In some embodiments, compositions of the present disclosure are applied to a
processed plant product.
In some embodiments, compositions of the present disclosure are applied to a
harvest plant or plant part at a
processing/shipping facility.
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In some embodiments, compositions of the present disclosure are applied to a
processed plant product.
The present disclosure thus extends to plants and plant parts that have been
treated with a composition of the
present disclosure (e.g., plant propagation materials coated with a
formulation comprising one or more enzymes of the
present disclosure, plants sprayed with a formulation comprising one or more
enzymes of the present disclosure, harvested
plant parts coated with a formulation comprising one or more enzymes of the
present disclosure), to plants grown from
plant propagation materials that were treated with a composition of the
present disclosure, to plant parts harvested from
plants that have been treated with a composition of the present disclosure, to
plant parts harvested from plants grown from
plant propagation materials that were treated with a composition of the
present disclosure, to crops comprising a plurality
of plants that were treated with a composition of the present disclosure, to
crops comprising a plurality of plants grown
from plant propagation materials that were treated with a composition of the
present disclosure, to crops treated with a
composition of the present disclosure, to processed products derived from
plants that were treated with a composition of
the present disclosure, to processed products derived from plants grown from
plant parts that were treated with a
composition of the present disclosure, and to processed products treated with
a composition of the present disclosure.
As noted above, proteins of the present disclosure may be formulated into
compositions comprising a variety of
components, such as adhesives (stickers), chemical actives, dispersants
(spreaders). drying agents, emulsifiers, microbes,
nutrients, pest attractants and feeding stimulants, pH control components,
postharvest treatments, rain fasteners,
rhealogical agents, safeners, stabilizers, UV protectants and wetting agents.
It is to be understood that compositions and
methods of the present disclosure may likewise be used in combination with
such components as separate and distinct
compositions (as part of an integrated pest management strategy. for example).
It is to be understood that compositions and methods of the present disclosure
are not limited to horticultural
uses. The same activities that make enzymes, fommlations, nucleic acids and
organisms of the present disclosure useful
for preventing, treating, suppressing, eliminating and/or reducing the
severity of infestations/infections of/by
horticultural pests likewise render them useful for preventing, treating,
suppressing, eliminating and/or reducing the
severity of infestations/infections of/by bacteria, fungi, insects and
oomycetcs in and on various media, such as food
storage containers, animal bedding/feed, clothing, hard surfaces, medical
instruments, etc. Thus, it is to be understood
that compositions and methods of the present disclosure may be modified for
use in any other industry or endeavor in
which such prevention, treatment, suppression, elimination and/or reduction in
disease severity may be of benefit.
The following is a non-exhaustive listing of concepts and embodiments
encompassed by the present disclosure:
Use of a protein or formulation of the present disclosure for any one, two,
three, four, five, six, seven, eight, nine, ten or
more of the following:
1) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a horticultural pest
2) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by an acarid
3) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a bacterium
4) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a fungus
5) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a gastropod
6) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by an insect
7) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a nematode
8) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by an oomycete
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9) preventing/treating/suppressing/eliminating an infestation/infection of
a plant or plant part by a protozoan
10) reducing disease severity in plants affected by one or more
horticultural pests
11) reducing disease severity in plants affected by a phytopathogenic
acarid
12) reducing disease severity in plants affected by a phytopathogenic
bacteria
13) reducing disease severity in plants affected by a phytopathogenic
fungus
14) reducing disease severity in plants affected by a phytopathogenic
gastropod
15) reducing disease severity in plants affected by a phytopathogenic
insect
16) reducing disease severity in plants affected by a phytopathogenic
nematode
17) reducing disease severity in plants affected by a phytopathogenic
oomycete
18) reducing disease severity in plants affected by a phytopathogenic
protozoa
19) treating a surface/substance that is susceptible to
infestation/infection by a horticultural pest
20) treating a surface/substance that is susceptible to
infestation/infection by an acarid
21) treating a surface/substance that is susceptible to
infestation/infection by a bacterium
22) treating a surface/substance that is susceptible to
infestation/infection by a fungus
23) treating a surface/substance that is susceptible to
infestation/infection by a gastropod
24) treating a surface/substance that is susceptible to
infestation/infection by an insect
25) treating a surface/substance that is susceptible to
infestation/infection by a nematode
26) treating a surface/substance that is susceptible to
infestation/infection by an oomycete
27) treating a surface/substance that is susceptible to
infestation/infection by a protozoan
28) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by a horticultural pest
29) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by an acarid
30) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by a bacterium
31) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by a fungus
32) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth systcm, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by a gastropod
33) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by an insect
34) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
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container, a postharvest treatment chamber or a postharvest shipping
container, by a nematode
35) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by an oomycete
36) preventing/treating/suppressing/eliminating an infestation/infection of
a container, such as a seed container, a
planting pot, a hydroponic growth system, an experimental growth chamber, a
greenhouse, a postharvest storage
container, a postharvest treatment chamber or a postharvest shipping
container, by a protozoan
37) treating (e.g., coating, dipping, drenching, fogging, misting, soaking,
spraying) a plant or plant part
38) treating (e.g., drenching, fogging, irrigating, misting, spraying) a
plant growth medium
39) treating a planting/irrigation/fertilization/harvesting/packing/storage
apparatus
40) treating (e.g., coating, dipping, drenching, fogging, misting, soaking,
spraying) a storage container prior
to/concurrently with/subsequent to introduction of a plant or plant part into
said storage container
41) treating (e.g., coating, dipping, drenching, fogging, misting, soaking,
spraying) a harvested plant or plant part
42) prolonging the shelf-life of a harvested plant or plant part
43) delaying the ripening of a harvested plant or plant part
44) hastening the ripening of a harvested plant or plant part
45) improving the efficacy of a chemical pesticide
46) improving the efficacy of a chemical acaricide
47) improving the efficacy of a chemical bactericide
48) improving the efficacy of a chemical fungicide
49) improving the efficacy of a chemical gastropodicide
50) improving the efficacy of a chemical herbicide
51) improving the efficacy of a chemical insecticide
52) improving the efficacy of a chemical nematicide
53) improving the efficacy of a chemical oomyceticide
54) improving the efficacy of a chemical protozoacide
55) improving the efficacy of a biological pesticide
56) improving the efficacy of a biological acaricide
57) improving the efficacy of a biological bactericide
58) improving the efficacy of a biological fungicide
59) improving the efficacy of a biological gastropodicide
60) improving the efficacy of a biological herbicide
61) improving thc cfficacy of a biological insecticide
62) improving the efficacy of a biological nematicide
63) improving the efficacy of a biological oomyceticide
64) improving the efficacy of a biological protozoacide
65) improving the efficacy of a preharvest treatment
66) improving the efficacy of a postharvest treatment
67) expanding the target spectrum of a chemical pesticide
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68) expanding the target spectrum of a chemical acaricide
69) expanding the target spectrum of a chemical bactericide
70) expanding the target spectrum of a chemical fungicide
71) expanding the target spectrum of a chemical gastropodicide
72) expanding the target spectrum of a chemical herbicide
73) expanding the target spectrum of a chemical insecticide
74) expanding the target spectrum of a chemical nematicide
75) expanding the target spectrum of a chemical oomyceticide
76) expanding the target spectrum of a chemical protozoacide
77) expanding the target spectrum of a biological pesticide
78) expanding the target spectrum of a biological acaricide
79) expanding the target spectrum of a biological bactericide
80) expanding the target spectnum of a biological fungicide
81) expanding the target spectrum of a biological gastropodicide
82) expanding the target spectrum of a biological herbicide
83) expanding the target spectrum of a biological insecticide
84) expanding the target spectrum of a biological nematicide
85) expanding the target spectrum of a biological oomyceticide
86) expanding the target spectrum of a biological protozoacide
87) expanding the target spectrum of a preharvest treatment
88) expanding the target spectrum of a postharvest treatment
89) reducing chemical pesticide-induced pest resistance and/or
phytotoxicity
90) reducing biological pesticide-induced pest resistance and/or
phytotoxicity
91) inclusion as part of an integrated pest management strategy.
Any of the foregoing uses in which any one, two, three, four, five, six,
seven, eight, nine, ten or more of the following is
true:
1) the protein is an enzyme
2) the protein is an enzyme belonging to EC 1
3) the protein is an enzyme belonging to EC 1.1
4) the protein is an enzyme belonging to EC 1.1.3
5) the protein is an enzyme belonging to EC 1.1.3.4
6) the protein is an enzyme belonging to EC 1.1.3.25
7) the protein is an enzyme belonging to EC 1.10
8) the protein is an enzyme belonging to EC 1.10.3
9) the protein is an enzyme belonging to EC 1.10.3.2
10) the protein is an enzyme belonging to EC 1.11
11) the protein is an enzyme belonging to EC 1.11.1
12) the protein is an enzyme belonging to EC 1.11.1.6
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13) the protein is an enzyme belonging to EC 1.11.1.7
14) the protein is an enzyme belonging to EC 1.14
15) the protein is an enzyme belonging to EC 1.14.99
16) the protein is an enzyme belonging to EC 1.14.99.56
17) the protein is an enzyme belonging to EC 3
18) the protein is an enzyme belonging to EC 3.1
19) the protein is an enzyme belonging to EC 3.1.1
20) the protein is an enzyme belonging to EC 3.1.1.3
21) the protein is an enzyme belonging to EC 3.1.1.5
22) the protein is an enzyme belonging to EC 3.1.1.11
23) the protein is an enzyme belonging to EC 3.1.1.32
24) the protein is an enzyme belonging to EC 3.1.4
25) the protein is an enzyme belonging to EC 3.1.4.3
26) the protein is an enzyme belonging to EC 3.1.4.11
27) the protein is an enzyme belonging to EC 3.2
28) the protein is an enzyme belonging to EC 3.2.1
29) the protein is an enzyme belonging to EC 3.2.1.1
30) the protein is an enzyme belonging to EC 3.2.1.3
31) the protein is an enzyme belonging to EC 3.2.1.4
32) the protein is an enzyme belonging to EC 3.2.1.5
33) the protein is an enzyme belonging to EC 3.2.1.6
34) the protein is an enzyme belonging to EC 3.2.1.7
35) the protein is an enzyme belonging to EC 3.2.1.8
36) the protein is an enzyme belonging to EC 3.2.1.11
37) the protein is an enzyme belonging to EC 3.2.1.14
38) the protein is an enzyme belonging to EC 3.2.1.15
39) the protein is an enzyme belonging to EC 3.2.1.17
40) the protein is an enzyme belonging to EC 3.2.1.21
41) the protein is an enzyme belonging to EC 3.2.1.37
42) the protein is an enzyme belonging to EC 3.2.1.39
43) the protein is an enzyme belonging to EC 3.2.1.41
44) the protein is an enzyme belonging to EC 3.2.1.55
45) the protein is an enzyme belonging to EC 3.2.1.58
46) the protein is an enzyme belonging to EC 3.2.1.59
47) the protein is an enzyme belonging to EC 3.2.1.73
48) the protein is an enzyme belonging to EC 3.2.1.75
49) the protein is an enzyme belonging to EC 3.2.1.78
50) the protein is an enzyme belonging to EC 3.2.1.91
51) the protein is an enzyme belonging to EC 3.2.1.101
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52) the protein is an enzyme belonging to EC 3.2.1.109
53) the protein is an enzyme belonging to EC 3.2.1.132
54) the protein is an enzyme belonging to EC 3.2.1.133
55) the protein is an enzyme belonging to EC 3.2.1.176
56) the protein is an enzyme belonging to EC 3.4
57) the protein is an enzyme belonging to EC 3.4.11
58) the protein is an enzyme belonging to EC 3.4.11.1
59) the protein is an enzyme belonging to EC 3.4.21
60) the protein is an enzyme belonging to EC 3.4.21.19
61) the protein is an enzyme belonging to EC 3.4.21.62
62) the protein is an enzyme belonging to EC 3.4.24
63) the protein is an enzyme belonging to EC 3.4.24.28
64) the protein is an enzyme belonging to EC 3.5
65) the protein is an enzyme belonging to EC 3.5.1
66) the protein is an enzyme belonging to EC 3.5.1.1
67) the protein is an enzyme belonging to EC 4
68) the protein is an enzyme belonging to EC 4.2
69) the protein is an enzyme belonging to EC 4.2.2
70) the protein is an enzyme belonging to EC 4.2.2.2
71) the protein comprises one or more polypeptides having about/at least
60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, '71,
72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,
91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 %
sequence identity to one or more of SEQ ID NOs: 1-48 and 98-150 or a mature
polypeptide thereof
72) the protein comprises one or more polypeptides encoded by a
polynucleotide having about/at least 60, 61, 62, 63,
64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % sequence identity to any one or more of SEQ ID
NOs: 49-97 and 151-203 or the
cDNA sequence thereof
73) the protein comprises one or more polypeptides derived from any one of
SEQ ID NOs: 1-48 and 98-150 by
substitution, deletion or insertion of one or more amino acids
74) the protein comprises one or more polypeptides derived from a mature
polypeptide of any one of SEQ ID NOs: 1-
48 and 98-150 by substitution, deletion or insertion of one or more amino
acids
75) the protein comprises one or more polypeptides derived from any one of
71) through 74) wherein the N- and/or C-
terminal end has been extended by the addition of one or more amino acids
76) the protein comprises a fragment of any one of 71) through 75)
77) the protein is an enzymatically active fragment/mutant/variant of any
one of SEQ ID NOs: 1-48 and 98-150 or a
mature polypeptide thereof
78) the formulation comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more proteins
of the present disclosure or enzymatically
active fragments/mutations/variants thereof
79) the formulation comprises one or more adhesives
80) the formulation comprises one or more pest attractants
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81) the formulation comprises one or more pest feeding stimulants
82) the formulation comprises one or more chemical pesticides
83) the formulation comprises one or more chemical acaricides
84) the formulation comprises one or more chemical bactericides
85) the formulation comprises one or more chemical fungicides
86) the formulation comprises one or more chemical gastropodicides
87) the formulation comprises one or more chemical herbicides
88) the formulation comprises one or more chemical insecticides
89) the formulation comprises one or more chemical nematicides
90) the formulation comprises one or more chemical oomyceticides
91) the formulation comprises one or more chemical protozoacides
92) the formulation comprises one or more biological pesticides
93) the formulation comprises one or more biological acaricides
94) the formulation comprises one or more biological bactericides
95) the formulation comprises one or more biological fungicides
96) the formulation comprises one or more biological gastropoclicides
97) the formulation comprises one or more biological herbicides
98) the formulation comprises one or more biological insecticides
99) the formulation comprises one or more biological nematicides
100) the formulation comprises one or more biological oomyceticides
101) the formulation comprises one or more biological protozoacides
102) the formulation comprises one or more dispersants
103) the formulation comprises one or more polyols
104) the formulation comprises glycerol
105) the formulation comprises sorbitol
106) the formulation comprises one or more pH control components
107) the formulation comprises an acetate buffer
108) the formulation comprises a sodium acetate buffer
109) the formulation comprises a carbonate buffer
110) the formulation comprises a sodium carbonate buffer
111) the formulation comprises a citrate buffer
112) the formulation comprises a sodium citrate buffer
113) thc formulation compriscs a phosphate buffer
114) the formulation comprises a potassium phosphate buffer
115) the formulation has a pH of about 3 to about 7
116) the formulation has a pH of about 3 to about 6
117) the formulation has a pH of about 3 to about 5
118) the formulation has a pH of about 4 to about 5
119) the formulation has a pH of less than 5
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120) the formulation has a pH of more than 5
121) the formulation has a pH of about 5 to about 6
122) the formulation has a pH of about 5 to about 7.5
123) the formulation has a pH of less than 6
124) the formulation has a pH of more than 6
125) the formulation has a pH of about 6 to about 7
126) the formulation has a pH of less than 6.5
127) the formulation has a pH of more than 6.5
128) the formulation has a pH of less than 7
129) the formulation has a pH of more than 7
130) the formulation has a pH of about 6 to about 7.5
131) the formulation has a pH of more than 7.5
132) the formulation has a pH of about 7.5 to about 10
133) the formulation has a pH of about 8 to about 10
134) the formulation has a pH of about 8.5 to about 9.5
135) the formulation comprises one or more rain fasteners
136) the formulation comprises one or more organo-modified siloxanes
137) the formulation comprises one or more organo-modified trisiloxanes
138) the formulation comprises one or more polyether trisiloxanes
139) the formulation comprises one or more organo-modified poly siloxanes
140) the formulation comprises one or more polyether polysiloxanes
141) the formulation comprises one or more preservatives
142) the formulation comprises potassium benzoate
143) the formulation comprises potassium sorbatc
144) the formulation comprises sodium benzoate
145) the formulation comprises sodium sorbate
146) the formulation comprises one or more postharvest treatments
147) the formulation comprises one or more essential oils
148) the formulation comprises one or more ethylene biosynthesis inhibitors
149) the formulation comprises one or more cyclopropenes
150) the formulation comprises one or more waxes
151) the protein or formulation is applied in a pesticidally effective amount
152) the protein or formulation is applied in an acaricidally effective amount
153) the protein or formulation is applied in a bactericidally effective
amount
154) the protein or formulation is applied in a fungicidally effective amount
155) the protein or formulation is applied in a gastropodicidally effective
amount
156) the protein or formulation is applied in an herbicidally effective amount
157) the protein or formulation is applied in an insecticidally effective
amount
158) the protein or formulation is applied in a nematicidally effective amount
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159) the protein or formulation is applied in an oomyceticidally effective
amount
160) the protein or formulation is applied in a protozoacidallv effective
amount
161) the protein or formulation is applied in a pesticidally ineffective
amount (e.g., as an adjuvant)
162) the protein or formulation is applied in an acaricidally ineffective
amount (e.g., as an adjuvant)
163) the protein or formulation is applied in a bactericidally ineffective
amount (e.g., as an adjuvant)
164) the protein or formulation is applied in a fungicidally ineffective
amount (e.g., as an adjuvant)
165) the protein or formulation is applied in a gastropodicidally ineffective
amount (e.g., as an adjuvant)
166) the protein or formulation is applied in a herbicidially ineffective
amount (e.g., as an adjuvant)
167) the protein or formulation is applied in an insecticidally ineffective
amount (e.g., as an adjuvant)
168) the protein or formulation is applied in a nematicidally ineffective
amount (e.g., as an adjuvant)
169) the protein or formulation is applied in an oomyceticidally ineffective
amount (e.g., as an adjuvant)
170) the protein or formulation is applied in a protozoacidally ineffective
amount (e.g., as an adjuvant)
171) the protein or formulation is used in combination with one or more
adhesives (e.g., in a tank mix)
172) the protein or formulation is used in combination with one or more pest
attractants (e.g., in a tank mix)
173) the protein or formulation is used in combination with one or more pest
feeding stimulants (e.g., in a tank mix)
174) the protein or formulation is used in combination with one or more
chemical pesticides (e.g., in a tank mix)
175) the protein or formulation is used in combination with one or more
chemical acaricides (e.g., in a tank mix)
176) the protein or formulation is used in combination with one or more
chemical bactericides (e.g., in a tank mix)
177) the protein or formulation is used in combination with one or more
chemical fungicides (e.g., in a tank mix)
178) the protein or formulation is used in combination with one or more
chemical gastropodicides (e.g., in a tank mix)
179) the protein or formulation is used in combination with one or more
chemical herbicides (e.g., in a tank mix)
180) the protein or formulation is used in combination with one or more
chemical insecticides (e.g., in a tank mix)
181) the protein or formulation is used in combination with one or more
chemical nematicides (e.g., in a tank mix)
182) the protein or formulation is used in combination with one or more
chemical oomyceticides (e.g., in a tank mix)
183) the protein or formulation is used in combination with one or more
chemical protozoacides (e.g., in a tank mix)
184) the protein or formulation is used in combination with one or more
microbial pesticides (e.g., in a tank mix)
185) the protein or formulation is used in combination with one or more
microbial acaricides (e.g., in a tank mix)
186) the protein or formulation is used in combination with one or more
microbial bactericides (e.g., in a tank mix)
187) the protein or formulation is used in combination with one or more
microbial fungicides (e.g., in a tank mix)
188) the protein or formulation is used in combination with one or more
microbial gastropodicides (e.g., in a tank mix)
189) the protein or formulation is used in combination with one or more
microbial herbicides (e.g., in a tank mix)
190) the protein or formulation is used in combination with one or more
microbial insecticides (e.g., in a tank mix)
191) thc protein or formulation is used in combination with one or more
microbial ncmaticidcs (e.g., in a tank mix)
192) the protein or formulation is used in combination with one or more
microbial oomyceticides (e.g., in a tank mix)
193) the protein or formulation is used in combination with one or more
microbial protozoacides (e.g., in a tank mix)
194) the protein or formulation is used in combination with one or more
diazotrophs (e.g., in a tank mix)
195) the protein or formulation is used in combination with one or more
phosphate-solubilizing microorganisms (e.g.,
in a tank mix)
196) the protein or formulation is used in combination with one or more plant
growth regulators (e.g., in a tank mix)
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197) the protein or formulation is used in combination with one or more rain
fasteners (e.g., in a tank mix)
198) the protein or formulation is used in combination with one or more organo-
modified siloxanes (e.g., in a tank mix)
199) the protein or formulation is used in combination with one or more organo-
modified trisiloxanes (e.g., in a tank
mix)
200) the protein or formulation is used in combination with one or more
polyether trisiloxanes (e.g., in a tank mix)
201) the protein or formulation is used in combination with one or more organo-
modified polysiloxanes (e.g., in a tank
mix)
202) the protein or formulation is used in combination with one or more
polyether polysiloxanes (e.g., in a tank mix)
203) the protein or formulation is used in combination with one or more
preservatives (e.g., in a tank mix)
204) the protein or formulation is used in combination with one or more
preharvest treatments (e.g., in a tank mix)
205) the protein or formulation is used in combination with one or more
postharvest treatments (e.g., in a tank mix)
206) the protein or formulation is used in combination with one or more
essential oils (e.g., in a tank mix)
207) the protein or formulation is used in combination with one or more
ethylene biosynthesis inhibitors (e.g., in a tank
mix)
208) the protein or formulation is used in combination with one or more
cyclopropenes (e.g., in a tank mix)
209) the protein or formulation is used in combination with one or more waxes
(e.g., in a tank mix).
A transgenic microorganism, plant or plant part that:
1) comprises one or more polynucleotides encoding a polypeptide
having about/at least 60, 61, 62, 63, 64, 65, 66, 67,
68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86,
87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,
98, 99 or 100 % sequence identity to one or more of SEQ ID NOs: 1-48 and 98-
150 or a mature polypeptide
thereof
2) comprises one or more polynucleotides having about/at least 60,
61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,
74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,
93, 94, 95, 96, 97, 98, 99 or 100 % sequence
identity to any one or more of SEQ ID NOs: 49-97 and 151-203 or the cDNA
sequence thereof
3) expresses one or more polypeptides having about/at least 60, 61,
62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,
75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,
94, 95, 96, 97, 98, 99 or 100 % sequence
identity to one or more of SEQ ID NOs: 1-48 and 98-150 or a mature polypeptide
thereof
4) comprises one or more polynucleotides encoding a polypeptide
derived from any one of SEQ ID NOs: 1-48 and
98-150 by substitution, deletion or insertion of one or more amino acids
5) expresses one or more polypeptides derived from any one of SEQ ID
NOs: 1-48 and 98-150 by substitution,
deletion or insertion of one or more amino acids
6) comprises one or morc polynucleotides encoding a polypeptide
derived from a mature polypeptide of any one of
SEQ ID NOs: 1-48 and 98-150 by substitution, deletion or insertion of one or
more amino acids
7) expresses one or more polypeptides derived from a mature
polypeptide of any one of SEQ ID NOs: 1-48 and
98-150 by substitution, deletion or insertion of one or more amino acids
8) comprises one or more polynucleotides encoding a polypeptide
derived from any one of any one of 71) through
74) above wherein the N- and/or C-terminal end has been extended by the
addition of one or more amino acids
9) expresses one or more polypeptides derived from any one of 71)
through 74) above wherein the N- and/or C-
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terminal end has been extended by the addition of one or more amino acids
10) comprises one or more polynucleotides encoding an enzymatically active
fragment of any one of 71) through 77)
above
11) expresses an enzymatically active fragment of any one of 71) through
77) above.
Use of a rain fastener for any one, two, three, four, five, six, seven, eight,
nine, ten or more of the following:
1) improving the adhesion/spreading/rainfastness of a protein on a
smface/substance that is susceptible to
infestation/infection by a horticultural pest
2) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by an acarid
3) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by a bacterium
4) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by a fungus
5) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by a gastropod
6) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by an insect
7) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by a nematode
8) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by an oomycete
9) improving the adhesion/spreading/rainfastness of a protein on a
surface/substance that is susceptible to
infestation/infection by a protozoan
10) improving the adhesion/spreading/rainfastness of a protein on a
container, such as a seed container, a planting pot,
a hydroponic growth system, an experimental growth chamber, a greenhouse, a
postharvest storage container, a
postharvest treatment chamber or a postharvest shipping container
11) improving the adhesion/spreading/rainfastness of a protein on a plant
or plant part
12) improving the adhesion/spreading/rainfastness of a protein on a
harvested plant or plant part
13) improving the pesticidal efficacy of a protein or formulation of the
present disclosure
14) improving the acaricidal efficacy of a protein or formulation of the
present disclosure
15) improving the bactericidal efficacy of a protein or formulation of the
present disclosure
16) improving the fungicidal efficacy of a protein or formulation of the
present disclosure
17) improving the gastropodicidal efficacy of a protein or formulation of
the present disclosure
18) improving the herbicidal efficacy of a protein or formulation of the
present disclosure
19) improving the insecticidal efficacy of a protein or formulation of the
present disclosure
20) improving the nematicidal efficacy of a protein or formulation of the
present disclosure
21) improving the oomyceticidal efficacy of a protein or formulation of the
present disclosure
22) improving the protozoacidal efficacy of a protein or formulation of the
present disclosure.
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Any of the foregoing uses in which any one, two, three, four, five, six,
seven, eight, nine, ten or more of the following is
true:
1) the rain fastener comprises one or more organo-modified siloxanes
2) the rain fastener comprises one or more organo-modified trisiloxanes
3) the rain fastener comprises one or more polyether trisiloxanes
4) the rain fastener comprises one or more trisiloxane ethoxylates
5) the rain fastener comprises one or more trisiloxane polyethoxylates
6) the rain fastener comprises one or more organo-modified polysiloxanes
7) the rain fastener comprises one or more polyether polysiloxanes
8) the rain fastener comprises one or more polysiloxane ethoxylates
9) the rain fastener comprises one or more poly siloxane poly ethoxy lates
10) the rain fastener comprises one or more organo-modified siloxanes
selected from the group consisting of
trisiloxanes and polysiloxanes described by the general Formula I:
R.13SiOIR.12SiOlAIR1R2SiO]ssa,3
wherein
A is 0-200, preferably A is 0-1, more preferably A is 0;
B is 0-200, preferably B is 0.5-2; more preferably B is 1;
A+B >0;
represents identical or different from each other hydrocarbon substituents of
1-10 carbons or hydrogen,
preferably methyl, ethyl, propyl and/or phenyl substituents, more preferably
methyl substituents; and
R2 represents identical or different from each other polyether substituents of
the general Formula II:
-RIOICH2CH201c[CH2CH(CH3)01D[CHR4CHR401ER5
wherein
R3 represents identical or different from each other hydrocarbon moieties of 1-
8 carbons, which optionally is
interrupted by oxygen atoms, preferably linear hydrocarbons of 2-4 carbons,
more preferably -CH2-CH2-CH2-;
10 represents identical or different from each other hydrocarbon substituents
of 1-12 carbons or hydrogen,
preferably methyl, ethyl, phenyl and/or hydrogen substituents;
R5 represents identical or different from each other hydrocarbon substituents
of 1-16 carbons, which optionally
contains urethane, carbonyl or carboxylic acid functionality, or hydrogen;
preferably methyl or hydrogen; more
preferably hydrogen;
C is 0-60, preferably C is 1-15;
D is 0-60, preferably D is 0-10;
E is 0-20, preferably E is 0-10, more preferably E is 0; and
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C+D+E >0
11) the rain fastener comprises Oxirane, 2-methyl-, polymer with oxirane,
mono [341,3,3,3-tetramethy1-1-
[(trimethylsilyl)oxy J-1-disiloxanyl]propyl] ether
12) the rain fastener comprises 3-(2-methoxyethoxy)propyl-methyl-
bis(trimethylsilyloxy)silane
13) the rain fastener comprises polyalkyleneoxide silane and
isotridecylalcohol ethoxylate
14) the rain fastener comprises 3-
(polyoxyethylene)propylheptamethyltrisiloxane
15) the rain fastener comprises siloxane polyalkyleneoxide and fatty
alcohol (C10-12) ethoxylate propox-ylate
16) the rain fastener comprises Oxirane, 2-methyl-, polymer with oxirane,
mono [3-[1,3,3,3-tetramethy1-1-
Rtrimethylsilypoxy1-1-disiloxanyllpropyll ether.
EXAMPLES
The following examples are provided to illustrate certain embodiments and are
not to be construed as limiting
the inventive concepts described in the present disclosure.
Examples 1-3: Organo-modified Siloxanes Improve Enzyme Rainfastness on Plant
Surfaces
Commercial adjuvants tested:
BREAK-THRU S 301: Oxirane, 2-methyl-, polymer with oxirane, mono [3-[1,3,3,3-
tetramethy1-1-[(trimethylsily1)oxyl-
1-disiloxanyllpropyll ether (Evonik Operations Gmbh, Essen, Germany)
SILWET' L-77: 3-(2-methoxyethoxy)propyl-methyl-bis(trimethylsilyloxy)silane
(Momentive Inc, Waterford, NY,
USA)
SILWET HS-312: Polyalkylencoxide silanc and isotridecylalcohol ethoxylate
(Momentive Inc, Waterford, NY, USA)
SILWET 408: 3-(Polyoxyethylene)propylheptamethyltrisiloxane (Momentive Inc,
Waterford, NY, USA)
SILWET STIK 2: Siloxanc polyalkylencoxide and fatty alcohol (C10-12)
ethoxylate propoxylatc (Momentive Inc,
Waterford, NY, USA)
SILWET 719: Oxirane, 2-methyl-, polymer with oxirane, mono p-[1,3,3,3-
tetramethy1-1-[(trimethylsilypoxy1-1-
disiloxanyllpropyll ether (Momentive Inc, Waterford, NY, USA)
Tween L-1010: Polyoxyethylene (10) polyoxypropylene (10) sorbitan monolaurate
(Croda International Plc, East
Yorkshire, United Kingdom)
Tweerri3) 24: Polyoxyethlene (16) sorbitan monolaurate (Croda International
Plc, East Yorkshire, United Kingdom)
ADSEETm 900: fatty alcohol ethoxylate (CIO) (Nouryon Surface Chemistry AB,
Stenungsund, Sweden)
ADSEETm 973: Alcohol ethoxylate blend (Nouryon Surface Chemistry AB,
Stenungsund, Sweden)
LUCROPO ROIL: Blend of cstcrficd oils and surfactants (LEVACO Chemicals Gmbh,
Leverkusen, Germany)
ADSEETM ST4: Maltodextrin/viaylpyrrolidone polymer (Nouryon Surface Chemistry
AB, Stenungsund, Sweden)
Ethylan 995: Alcohol ethoxylate propoxylate (C16-18) (Nouryon Surface
Chemistry AB, Stenungsund, Sweden)
Ethomeen C/15: Coco alkylamine ethoxylate (Nouryon Surface Chemistry AB,
Stenungsund, Sweden)
Rain fastness assay:
The rain fastness assay was performed in a 24 well plate. The wells were
filled with a heated 1% agar solution
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that solidifies after cooling to room temperature. 12 mm Leaf discs of
preferred plants were prepared and added on top of
the agar (the agar supported the leaf to not dry out so quickly).
After preparation of tank mix-formulations a 25 I, droplet of each sample
were put in the middle of the leaf
disc. The spreading behavior of the droplet was observed to avoid including
results of samples in which the droplet
spread so far that material was lost from the leaf. After addition of all
samples the well plate was left to dry in a fume
hood for 24h.
When dried, each disc was washed with 1 mL of tap water. The water was
dispensed onto the leaf a total of 3
times. Afterwards the sample was analyzed for enzyme activity to determine
residual amount on the leaf by using
following equation:
% Retention of treatment = 1 ¨ (activity units enzyme in wash solution /
activity units spray-solution
added onto leaf)
The amount of protein added to spray solutions was given as weight percent (%
w/w) protein of interest, which in the
case of enzymes was determined using a colorimetric assay.
EXAMPLE 1
The retention of three enzymes¨a cellobiose oxidase obtained from Microdochlum
'male (NZ protein # 6;
SEQ ID NO: 6), a catalase obtained from Aspergillus niger (NZ protein # 10;
SEQ ID NO: 10), and a serine protease
obtained from Nocardiopsis sp. (NZ protein # 42; SEQ ID NO: 42)¨with and
without a trisiloxane adjuvant, BREAK-
THRU S 301, was evaluated. The spray-solutions were applied on top of tomato
and grape leaf discs. Table 2 shows the
retention of enzyme on the leaf after a simulated rain event.
Table 2. Retention of proteins on tomato and grape leaves after a simulated
rain event.
Foliar Spray Formulation Leaf % Retention of
treatment
0.0059% NZ Protein # 42
0.95% K2HPO4
Tomato 40%
0.05% KH2PO4
pH 7.9
0.0059% NZ Protein # 42
0.05% BREAK-THRUO S 301
0.95% K2HPO4 Tomato 95%
0.05% KH2PO4
pH 7.9
0.0059% NZ Protein # 42 Grape 54%
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0.95% K2HPO4
0.05% KH2PO4
pH 7.9
0.0059% NZ Protein # 42
0 05% BREAK -THRT_T S 301
0.95% K2HPO4 Grape 100%
0.05% KH2PO4
pH 7.9
0.001% NZ Protein #6
0.6% K2HPO4
Grape 68%
0.4% KH2PO4
pH 6.9
0.001% NZ Protein #6
0.05% BREAK-THRU S 301
0.6% K2HPO4 Grape 95%
0.4% KH2PO4
pH 6.9
0.027% NZ Protein # 10
0.6% K2HPO4
Tomato 49%
0.4% KH2PO4
pH 6.9
0.027% NZ Protein # 10
0.05% BREAK -THRI S 301
0.6% K2HPO4 Tomato 79%
0.4% KH2PO4
pH 6.9
0.027% NZ Pmtein # 10
0.6% K2HPO4 Grape 62%
0.4% KH2PO4
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0.027% NZ Protein # 10
0.05% BREAK-THRUO S 301
0.6% K2HPO4 Grape 86%
0.4% KH2PO4
pH 6.9
The results in Table 2 show a clear difference in rainfastness when
formulating the spray tank solution with a
trisiloxanc (BREAK-THRU S301). Further, the results show that there is a
significant improvement in rainfastness,
independent of enzyme class and leaf type.
EXAMPLE 2
The retention of a serine protease obtained from Nocardiopsis sp. (NZ protein
# 42; SEQ ID NO: 42) on
tomato leaf discs was evaluated in presence of various commercially available
organo-modified siloxanes.
Table 3 shows the retention of protease in the rainfastness assay with the
tested organo-modified siloxanes.
Both trisiloxanes (SILWET L-77, SILWET 408, SILWET 719 and BREAK-THRU S
301) and polysiloxanes
(SILWET HS-312 and SILWET STIK 2) give rise to significantly improved
rainfastness of the protease.
Table 3. Retention of a serine protease obtained from Nocardiopsis sp. (NZ
protein # 42; SEQ ID NO: 42) on
tomato leaf discs in presence of different organo-modified siloxanes.
Foliar Spray Formulation A) Retention of treatment
0.0059% NZ Protein # 42 58%
0.0059% NZ Protein # 42
71%
0.1% SILWET Tm HS-312
0.0059% NZ Protein 1142
80%
0.1% SILWET 'M L-77
0.0059% NZ Protein # 42
94%
0.1% SILWET' 408
0.0059% NZ Protein # 42
93%
0.1% SILWETIM STIK 2
0.0059% NZ Protein # 42
92%
0.1% SILWET Tm 719
0.0059% NZ Protein 1142 91%
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0.1% BREAK-THRUk S 301
EXAMPLE 3
The retention a se rine protease obtained fro iii Nocurdiopsis sp. (NZ protein
# 42; SEQ ID NO: 42) on tomato
leaf discs after a simulated rain event in 24-well plates was determined when
combining the enzymes with a range of
commercially available adjuvants.
Table 4. Effect of common types of adjuvants on enzyme retention after
simulated rain
Formulation % Retention of treatment
0.0059% NZ Protein # 42 60%
0.0059% NZ Protein # 42
87%
0.1% BREAK-THRU S301
0.0059% NZ Protein # 42
42%
0.1% Tweenk L-1010
0.0059% NZ Protein # 42
38%
0.1% Tween 24
0.0059% NZ Protein # 42
38%
0.1% ADSEE 900
0.0059% NZ Protein # 42
20%
0.1% ADSEErm 973
0.0059% NZ Protein # 42
48%
0.1% LUCROP ROIL
0.0059% NZ Protein # 42
45%
0.1% ADSEETm ST4
0.0059% NZ Protein # 42
48%
0.1% ETHYLAN 995
0.0059% NZ Protein # 42
22%
0.1% ETHOMEEN C/15
The experimental data show the unique rainfastness improvement of enzymes seen
with organo-modified
siloxane, which cannot be observed for other adjuvants commonly used in
agricultural applications.
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Examples 4-5: Alkaline pH Improves the Fungicidal Efficacy of a Serine
Protease
EXAMPLE 4
The efficacy of a serine protease obtained from Nocardiopsis sp (NZ protein #
42; SEQ ID NO: 42) against
tomato late blight and cucumber downy mildew was tested at alkaline pH in a
plate-based assay.
For the tomato late blight plate assay. 24 well plates were lined with 2 mL of
1.2% agar supplemented with
0.5% sorbitol. Each well had a E125 cm leaf disc cut with a #6 cork borer from
tomato plants (cv. Moneymaker). The
leaf disc was placed adaxial side up (top side facing up). To each leaf disc
15 juL of our treatments for a total of 8
wells/replicates per treatment. Leaf discs were allowed to dry for 2.25 hours
in a laminar flow hood. Each leaf disc was
wounded before inoculating 10 !A., of the spore suspension at a concentration
of 5E4 sporangia/mL onto the wound. Each
plate was sealed with parafilm and held at 18 degrees C and 18:6 light:dark
cycle for 5 days before being scored (0 = no
infection, 1 = infection).
For the cucumber downy mildew plate assay 48 well plates were lined with 700
lit of 1% agar. Each well
contains a 0.875 cm leaf disc of cucumber (cv. Iznik) with the bottom side
facing up. To each well we add 15 L. of our
treatments for a total of 16 wells treatment. The discs were dried for 1-2
hours in a laminar flow hood. The pathogen was
prepared by scraping infected and sporulating cucumber leaves with a 10 jiL
loop submerged in sterile water, and then
the sporangia were quantified via a haemocytometen and finally adjusted to a
concentration of 5E4 sporangia/mL. Each
leaf disc was woundcd and then 10 [IL of a 5E4 sporangia/mL solution was
pipetted onto the wound. Plates were held at
18 degrees C and 18:6 light:dark cycle for 5 days before being scored (0 = no
infection, 1 = infection).
Table 5. Effect of pH on the effectiveness of a scrinc protease obtained from
Nocardiopsis sp. (NZ protein # 42;
SEQ ID NO: 42) against tomato late blight and cucumber downy mildew
Spray solution Protease dosage (mg Leaf type
Pathogen incidence
active enzyme/L spray
solution)
Water 0 Cucumber 100%
Water 6 Cucumber 92%
Water 60 Cucumber 67%
0.8% K2HPO4 0 Cucumber 94%
0.06% KH2PO4
pH 7.9
0.8% K2HPO4 6 Cucumber 63%
0.06% KH2PO4
pH 7.9
0.8% K2HPO4 60 Cucumber 21%
0.06% KILP04
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pH 7.9
Water 0 Tomato 94%
Water 6 Tomato 100%
Water 60 Tomato 100%
0.8% K2HPO4 0 Tomato 75%
0.06% KH2PO4
ph I 7.9
0.8% K2HPO4 6 Tomato 44%
0.06% KH2PO4
pH 7.9
0.9% K2HPO4 60 Tomato 25%
0.06% KH2PO4
pH 7.9
EXAMPLE 5
The efficacy of a serine protease obtained from Nocardiopsis sp (NZ protein #
42; SEQ ID NO: 42) was tested
at alkaline pH against tomato late blight in green house trials.
For the greenhouse assays, 30 day old tomatoes (cv. Sungold) were treated with
treatments at a rate of 30-40
gallons per acre and allowed to air dry for 24 hour before they were sprayed
with a sporangia suspension (1E4
sporangia/mL in sterile ddH20) of P. infestans.
After being sprayed with sporangia (approximately 10 ml. per plant) plants
were placed in the dark at 18
degrees C with a humidifier for 48 hours. Plants were then removed and kept at
18 degrees C with 16:8 light:dark cycle.
After 5 days the plants were assessed for disease severity (percentage of
disease tissue of the entire plant).
The disease severity measured on an untreated control was 32.5%.
Table 6. Effect of pH on the effectiveness of a scrinc protease obtained from
Nocardiopsis sp. (NZ protein # 42;
SEQ ID NO: 42) against tomato late blight
Treatment % Disease reduction compared to
untreated
control
Water 58%
Protease: 15 mg active enzyme/L
0.8% K2HPO4 98%
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0.06% KH2PO4
pH 7.9
0.1% S1LWETTm L-77
Protease: 15 mg active enzyme/L
0.9% K2HPO4 92%
pH 9
0.1% SILWETTm L-77
Protease: 15 mg active enzyme/L
Examples 6-9: Organo-Modified Trisiloxanes Improve the Retention of Enzymes
Sprayed on Plant Surfaces and
Their Effectiveness Against Zvmoseptoria tritici
Principle
A STB susceptible winter wheat cultivar (var. Hereford) growing in 1 L pots
was first sprayed with enzyme and after 24
hours sprayed with a spore suspension of Zymoseptoria tritici. Percent leaf
area attacked was assessed 14 days after
inoculation and again with 3 days interval until the full effects were found.
For scoring the standard EPPO scales were
used (EPPO standards: Guidelines on good plant protection practice. Wheat. PP
2/10(1)). Experiments were conducted by
Aarhus University, Flakkebjerg, Denmark.
Materials
Winter wheat (var. Hereford), Zymoseptoria tritici spore suspensions (see
Inoculum A and Inoculum B below), K2HPO4,
KH2PO4, sodium acetate, acetic acid, glycerol, Tween 20 (polyoxyethylene
sorbitol ester, Sigma-Aldrich), BREAK-
THRU* S 301 (poly ether trisiloxarie, Evonik Industries AG), and SILWETTm L77
(polyalkyleneoxide modified
heptamethyltrisiloxane, Momentive Performance Materials Inc.). Enzymes are
described in the examples
Buffers
1M potassium phosphate stock solutions, pH 6, 7 or 8, were prepared as shown
in Table 7 using dem ineralized water.
Table 7
Buffer K2PO4 (g) KH2PO4 (g) Total
volume
(mL)
pH 6 5.24 9.52 100
pH 7 11.20 4.85 100
pH 8 16.30 0.90 100
If needed, pH was adjusted to target value using a hydrochloric acid or sodium
hydroxide solution.
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1 M acetate, pH 5 stock solution was prepared by mixing 5.7 g sodium acetate
and 1.8 g glacial acetic acid with water
(total volume 100 inL) (pH 5 buffer).
Inoculums (Zymoseptoria hind spore suspension)
Inoculum A
Five isolates of Zymoseptoria tritici, collected in 2020 from naturally
infected plants in Danish fields and considered to
represent the natural Septoria community in Denmark that year, were grown on
PDA (Potato Dextrose Agar, available e.g.
from Sigma-Aldrich) and respective spores harvested and mixed into an unique
suspension with a concentration similar to
2 x 106 spores per ml. The inoculum was freshly prepared before use and
contained 0.1% Tweeng, 20 to ensure good
spreading on the leaves.
Inoculum B
Five isolates of Zymoseptoria tritici, collected in 2021 from naturally
infected plants in Danish fields and considered to
represent the natural Septoria community in Denmark that year, were grown on
PDA (Potato Dextrose Agar, available e.g.
from Sigma-Aldrich) and respective spores harvested and mixed into an unique
suspension with a concentration similar to
2 x 106 spores per ml. The inoculum was freshly prepared before use and
contained 0.1% Tweeng 20 to ensure good
spreading on the leaves.
Procedure
1) Wheat kernels were sown in 1 L pots and allowed to germinate and grow under
greenhouse conditions for
approximately 2 weeks until reaching the two-leaf crop growth stage (BBCH12).
2) A tank mix for each enzyme to be tested was prepared by diluting the buffer
stock solution 100-fold (resulting
concentration in tank mix was 10 mM) and adding the enzyme to the
concentration indicated in the examples and
surfactant to a final concentration of 0.1 % (w/v). The buffer pH, type of
surfactant and type of enzyme are given
in the examples. In some experiments no buffer was added.
3) The plants were treated by spraying the tank mix solution using a cabin
sprayer (volume ¨600 mL; 150 L/ha, 3.6
km/hour, yellow nozzles 0.2). Tank mix solutions were prepared a few hours
before application took place. The
pH of the buffer depended on the enzyme used.
4) For the Septoria inoculum, 5 isolates of Zymoseptoria tritici collected
from naturally infected plants in 2020
Danish fields were grown on PDA and the respective spores collected and mixed
into a solution. The Septoria
inoculum had a concentration similar to 2 x 10' sporcs/mL and contained 0.1 %
Tweent 20 to ensure that the
spores spread well on the leaves. The inoculum was freshly prepared before
use.
5) 24 hours after application of the enzyme, the plants were sprayed with the
Septoria inoculum (2 mL per pot).
After inoculation, the plants were kept under high relative humidity, covered
with black plastic for 3 days and
transparent plastic for additional 9 days to ensure infection and further
development of the pathogen.
6) 6 replicates were made for each condition. As control, plants were
sprayed with a tank mix prepared with 0.1 %
SILWETTmL-77 (no enzyme added).
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7) Percent leaf area attacked by STB was determined 14 days after
inoculation and again with 3 days interval until
the full effects were found. For scoring, the standard EPPO scale was used
(EPPO standards: Guidelines on good
plant protection practice. Wheat. PP 2/10(1)).
Example 6
Enzymes were tested for efficacy on Zymoseptoria tritici infected winter wheat
as described above using Inoculum A. The
enzymes tested, buffers and surfactants used as well as results obtained are
described in Table 8. Results are presented as
percent disease control in relation to the untreated control (UTC) at 18.7%
disease pressure. The higher the % control, the
better is the result. 100 % means no disease was detected in the treatment and
0% means disease as at the same level as
the untreated control (UTC). Tank mix with pH buffer and SILWETIm L-77
improved the performance of NZ protein
numbers 1 & 10, 1 (p = 0.153), 42 (p = 0.004), 21 (p = 0.014) and 22, as
compared to tank mix with Tween0 20.
Table 8. Effect of an organo-modified trisiloxane on the efficacy of enzymes
sprayed on wheat
Tank mix description
Enzyme Treatment
% Control
NZ Protein #(s) Enzyme conc.
Buffer surfactant
mg/mL
UTC SILWETTm L-77 0.0
1 & 10 0.034 Tween0 20
53.6
1 & 10 pH 7 0.034 SlLWETTm L-77
61.6
1 0.220 ** Tween 20
32.1
1 pH 6 0.220 ** SILWET L-77
64.3
42 0.068 Twee n* 20
33.0
42 pH 8 0.068 SILWETTm L-77
75.9
21 0.122 Tweenk 20
56.3
21 pH 5 0.122 SILWETTm L-77
79.5
22 0.038 Tweenk 20
34.8
22 pH 5 0.038 SILIVETTm L-77
50.9
** 0.1 inL glycerol per 100 mL tank mix was included
Example 7
Enzymes were tested for efficacy on Zymoseploria fr//ic/infected winter wheat
as described above using Inoculum A. The
enzymes tested, buffers and surfactants used as well as results obtained are
described in Table 9. Results are presented as
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percent disease control in relation to the untreated control (UTC) at 15.7%
disease pressure. The higher the % control, the
better is the result. 100 % means no disease was detected in the treatment and
0% means disease as at the same level as
the untreated control (UTC). Tank mix with pH buffer and SILWETTm L-77
significantly improved the performance of
the pectinase mixture (p = 0.069), as compared to tank mix with Tween 20.
Table 9. Effect of an organo-modified trisiloxane on the efficacy of enzymes
sprayed on wheat
Tank mix description
Enzyme Treatment
Enzyme conc. %
Control
NZ Protein #(s) Buffer surfactant
mg/mL
UTC SILWETTm L-77
0.0
pectinases 0.023 Tween 20 37.2
pectinases pH 5 0.023 SILWETrm L-77 61.7
45 0.051 Tween 20 76.1
45 pH 8 0.051 SILWETTm L-77 76.1
Example 8
Enzymes were tested for efficacy on Zymoseptoria tritici infected winter wheat
as described above using Inoculum B. The
enzymes tested, buffers and surfactants used as well as results obtained are
described in Table 10. Results are presented as
percent disease control in relation to the untreated control (UTC) at two
timepoints: Ti ¨ 19.2% disease pressure; T2 ¨
29.2% disease pressure. The higher the % control, the better is the result.
100 % means no disease was detected in the
treatment and 0% means disease as at the same level as the untreated control
(UTC). Tank mix with pH buffer and
SILWETTm L-77 improved the performance of NZ protein numbers 42, as compared
to tank mix with pH buffer and
BREAK-THRU S 301. Tank mix with pH buffer and BREAK-THRU S 301 improved the
performance of NZ protein
numbers 10, as compared to tank mix with pH buffer and SILWETTm L-77. Tank mix
with pH buffer and SILWETTm L-
77 improved the performance of NZ protein number 1, as compared to tank mix
with pH buffer and BREAK-THRUlt S
301 or Tween 20 alone. Tank mix with pH buffer and BREAK-THRUO S 301 also
improved the performance of NZ
protein number 1, as compared to tank mix with TWEEN 20 alone.
Table 10. Effect of organo-modified trisiloxanes on the efficacy of enzymes
sprayed on wheat
Enzyme Tank mix description
%Control
%Control
Treatment
Enzyme conc.
Buffer surfactant Ti
T2
NZ Protein #(s) mg/mL
UTC STLWETTm L-77 0.0
0.0
42 pH 8 0.068 SILWET m L-77 67.0
62.9
42 pH 8 0.068 SILWETTm L-77 53.9
74.3
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42 pH 8 0.068 BREAK-THRUg S 301 39.1 42.9
42 pH 8 0.017 SILWETTm L-77 80.9 82.3
42 pH 8 0.017 BREAK-THRUO S 301 54.8 55.4
10 pH 7 0.034 SILWETTm L-77 42.6 48.6
10 pH 7 0.034 BREAK-THRUCDi S 301 83.5
71.4
10 - 0.034 Tween 20 91.3 88.6
1 pH 6 0.220 ** SILWETTm L-77 89.6 84.0
1 pH 6 0.220 ** BREAK-THRUO S 301 84.3 77.7
1 pH 6 0.110 *** SILWETTm L-77 90.4 84.6
1 - 0.220 ** Tweeng 20 75.7 61.1
** 0.1 irriL glycerol per 100 iriL tank mix was included
*** 0.05 mL glycerol per 100 mL tank mix was included
Example 9
Enzymes were tested for efficacy on Zymoseptoria tritici infected winter wheat
as described above using Inoculum B. The
enzymes tested, buffers and surfactants used as well as results obtained are
described in Table 11. Results are presented as
percent disease control in relation to the untreated control (UTC) at two
timepoints: Ti - 16.7% disease pressure; T2 -
26.7% disease pressure. The higher the % control, the better is the result.
100 % means no disease was detected in the
treatment and 0% means disease as at the same level as the untreated control
(UTC). Tank mix with pH buffer and
SILWETTm L-77 improved the performance of NZ protein number 21, as compared to
tank mix with pH buffer and
BREAK-THRU CR) S 301 or TWEEN 20 alone. Tank mix with pH buffer and BREAK-
THRU (13.) S 301 also improved the
performance of NZ protein number 21, as compared to tank mix with TWEEN 20
alone. Tank mix with pH buffer and
SILWETTm L-77 improved the performance of NZ protein number 22, as compared to
tank mix with pH buffer and
BREAK-THRUO S 301. Tank mix with pH buffer and SILWETTm L-77 improved the
performance of NZ protein number
25, as compared to tank mix with TWEEN 20.
Table 11. Effect of organo-modified trisiloxanes on the efficacy of enzymes
sprayed on wheat
Tank mix description
Enzyme Treatment %Control
%Control
Enzyme conc.
NZ Protein #(s) Buffer surfactant Ti
T2
mg/mL
UTC - - SILWETTm L-77 0.0
0.0
21 pH 5 0.122 SILWETTm L-77 85.0
85.6
21 pH 5 0.122 BREAK-THRU S 301 79.0
70.6
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21 0.122 Tweenk 20 61.0
45.6
22 pH 5 0.038 STLWETTm L-77 87.0
84.4
22 pH 5 0.038 BREAK-THRUk S 301 70.0
65.6
25 pH 7 0.022 SILWETTm L-77 52.0
58.6
25 0.037 Tweenk 20 28.1
32.3
Example 10: Organo-Modified Trisiloxanes Improve the Retention of Enzymes
Sprayed on Plant Surfaces and
Their Effectiveness Against Phytophthora infestans
Principle
A late blight susceptible tomato variety (Sungo1d) was grown in the greenhouse
for approximately 3-weeks and then leaves
were harvested for leaf disk assays within 24-well plates. Leaf disks were
treated with 0.25 to 2.5 mg/ml enzyme and
allowed to diy fully for approximately 2 hours. Once dry, 10 [El of
Phytophthora h2festans (US 23) was pipetted onto the
leaf surface at a concentration of 1x105 sporangia per ml. Disease was
assessed 6 days post inoculation. Experiments were
conducted by Novozymes Biologicals, Salem, VA, USA.
Materials
o Susceptible tomato plants (Sungold)
o Rye A media (60 g Rye grains, 20 g sucrose, 15 g agar per liter)
o Enzyme(s)
o Phytophthora injestans (US 23 isolate)
o Potassium Phosphate Buffer (0.05 M, pH 7.88)
o Adjuvant A: SILWETTm L-77 (trisiloxane ethoxylate)
o Adjuvant B: BREAK-THRU SP 133 (polyglycerol esters and fatty acid
esters)
o 0.5% Butterfield's buffer agar (5 g agar per liter, 1 L Butterfield's
buffer), 2 ml within each well of a 24-well
plate
o 10 ml atomizer spray bottles
o Incubator (18 C) with cool fluorescent light on timer
o 14 mm Cork borer
o Hemocytometer
o Phase contrast microscope (10x objective)
Procedure
Leaf disk assay
1. 24-well plates containing 2m1 of solidified 0.5% Butterfield's buffer agar
(Butterfield's buffer was used as a
replacement to typical water agar to balance pH) were briefly dried within a
hood to remove condensation.
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2. Enzyme dilutions were made with potassium phosphate buffer (0.05 M, pH
7.88) with protein concentrations of one
of the following buffers and protein concentrations 0.25, 0.5, 1.0, and 2.5
mg/ml
3. In some instances, adjuvants were also applied with enzymes and their
appropriate buffers. The adjuvants and tested
concentrations are listed below:
SILWETTm L-77 at a concentration of 0.1%
BREAK-THRU SP 133 at a concentration of 0.1%
4. Leaves were detached fmm the mid-region of a susceptible tomato variety
(Sungold) at approximately 3-weeks of
age.
5. Leaf disk were excised using a cork borer (14 mm diameter) and placed
(adaxial/upper side up) into each of the
wells.
6. Enzyme dilutions were applied to the leaf surface with atomizer spray
bottles. Two spray pumps were applied to
each leaf disk which adequately covered the leaf disks without pool ing. Wells
with different treatments were covered
during application to eliminate contamination from overspray.
7. Enzyme applications were left to dry on the leaf surface for
approximately 2 hours.
8. While waiting, the spore solution was prepared using the protocol below.
9. After enzymes were thy, the center of the leaf disk was inoculated with
10 p.1 of the spore suspension containing
lx 105 sporangia per ml (or 10 p,1 sterile water for uninoculated controls).
10. Plates were sealed with parafilm and incubated at 18 C under a 16-hour
light/8-hour dark cycle.
11. Images of the leaf disk plates were captured and analyzed for disease at 5-
6 days post infection.
Spore Solution Preparation
1. Phytophthora infestans was grown on Rye A medium at 18 C under dark
conditions for 2-3 weeks.
2. Plates were flooded with approximately 3 ml cold sterile distilled water,
and the surface of the plate was gently
scraped with an inoculation loop.
3. Water was pipetted off the plate and placed into a 15 ml conical tube.
At roughly a 45-degree angle, another 2 ml
of sterile distilled water was pipetted one ml at a time over the plate. The
volume was transferred into the same
15m1 conical tube.
4. Sporangia were enumerated using a hemocytometer.
5. The spore suspension was incubated at 4 C for 0.5-2 hours to induce
zoospore release and then diluted to a
concentration of 1x105 sporangia per ml.
Example 10
Enzymes wcrc tested for efficacy against Phytophthora infestans infected
tomato as described above. In this example,
enzymes were diluted using potassium phosphate buffer (0.05 M, pH 7.88) and
tested at concentrations of 0.25, 0.5. 1.0,
and 2.5 mg protein per ml. Enzymes were tested without adjuvant as well as
with 0.1 % SILWETTm L-77 or 0.1 % BREAK-
THRU SP 133. Efficacy observed for disease control on the leaf disks at 6
days post infection is shown in Table 12. A
minimum of two experimental repeats with four replicates each were completed
for each treatment. Disease reduction is
defined as (-) no reduction, (+) mild reduction, (++) moderate reduction, and
(m) severe reduction. Dose responses can
be observed for the enzymes as well as improved disease reduction with the
addition of SILWETTm L-77 and BREAK-
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THRU SP 133. In some instances, addition of the adjuvant allowed for
comparable efficacy results at a lower enzyme
dosage.
Table 12. Effect of organo-modified trisiloxanes on the efficacy of enzymes
sprayed on tomato leaves
Enzyme Treatment Protein concentration
Disease
Adjuvant
NZ Protein #(s) (mg/10
Reduction
UTC 0 none -
UTC 0 SILWETTm L-77
+
UTC 0 BREAK-THRU SP 133 -
45 0.25 None -
45 0.50 None -
45 1.0 None -
45 2.5 None
45 0.25 SILWETTm L-77
+
45 0.50 SILWET' " L-77
+
45 1.0 SILWET' ' L-77
+++
45 2.5 SILWET' " L-77
+++
45 0.25 BREAK-THRU SP 133
+
45 0.50 BREAK-THRU SP 133
+
45 1.0 BREAK-THRU SP 133
+
45 2.5 BREAK-THRU SP 133
++
33 0.25 None -
33 0.50 None -
33 1.0 None
+
33 2.5 None
++
33 0.25 SILWETTm L-77
+
33 0.50 SILWETIm L-77
+
33 1.0 SILWETTm L-77
+
33 2.5 SILWETTm L-77
+++
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33 0.25 BREAK-THRU SP 133 -
33 0.50 BREAK-THRU SP 133 -
33 1.0 BREAK-THRU SP 133 -
33 2.5 BREAK-THRU SP 133
+++
43 0.25 None
+
43 0.50 None
+
43 1.0 None
+
43 2.5 None
++
43 0.25 SILWETTIvi L-77
+
43 0.50 SILWET' L-77
+
43 1.0 SILWET' L-77
++
43 2.5 SILWETim L-77
++
43 0.25 BREAK-THRU" SP 133 -
43 0.50 BREAK-THRU SP 133 -
43 1.0 BREAK-THRU SP 133
++
43 2.5 BREAK-THRU SP 133
+++
44 0.25 None
+
44 0.50 None
+
44 1.0 None
++
44 2.5 None
+++
44 0.25 SILWETTm L-77
+
44 0.50 SILWET' L-77
+
44 1.0 SILWET' " L-77
++
44 2.5 SILWET' L-77
+++
44 0.25 BREAK-THRU SP 133
+
44 0.50 BREAK-THRU SP 133
+
44 1.0 BREAK-THRU SP 133
44 2.5 BREAK-THRU SP 133
-Hp+
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