Note: Descriptions are shown in the official language in which they were submitted.
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Enzymatic Textile Colour Modification
The present application claims priority to U.S. Provisional Application Serial
No. 61/223,348,
filed on July 7, 2009, U.S. Provisional Application Serial No. 61/322,743,
filed on April 9, 2010,
European Patent Application No. 09162047.6, filed on June 5, 2009 and European
Patent
Application No. 09163751.2, filed on June 25, 2009, which are hereby
incorporated by
reference in their entirety.
The present invention relates to methods for the enzymatic colour modification
of dyed
cellulosic textile fibre material, in particular denim dyed with indigo or
sulphide dyes.
Some textile materials are washed after dyeing with the objective of adjusting
the colour tone
or shade on the dyed textile, also known as washdown effect. For instance,
blue jeans made
from indigo-dyed denim can be washed in the presence of pumice stones and
enzymatic
desizing agents, followed by an on tone-washdown process to obtain a desired
worn
appearance. Conventional washdown processes comprise treatment of the coloured
denim
with sodium hypochlorite, which is undesirable in view of the appearance of
fibre damages and
because of ecological reasons.
Washdown with hydrogen peroxide is an alternative solution. The adjusting
effect obtainable
with hydrogen peroxide, however, is rather limited. Furthermore, the required
high pH is
ecologically undesirable.
An enzymatic washdown process which does not show the above indicated
disadvantages
would be desirable.
There is a need for an effective enzymatic washdown process for coloured
cotton textiles
which provides the desired wash-out effect under mild conditions and minimizes
the adverse
environmental impact, when compared to conventional textile colour
modification processes.
The present invention accordingly relates to a method for adjusting the colour
tone of dyed
cellulosic textile fibre material comprising contacting said textile material
with an enzymatic
textile treatment composition comprising
(i) a perhydrolase enzyme,
(ii) an ester substrate for said perhydrolase enzyme, and
(iii) a hydrogen peroxide source.
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The enzymatic treatment step of the present invention will employ, unless
otherwise indicated,
conventional techniques of molecular biology (including recombinant
techniques),
microbiology, cell biology, and biochemistry, which are within the skill of
the art. Such
techniques are explained fully in the literature, for example, Molecular
Cloning: A Laboratory
Manual, 2nd ed., (Sambrook et al., 1989); Oligonucleotide Synthesis (M.J.
Gait, ed., 1984);
Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1994); PCR:
The
Polymerase Chain Reaction (Mullis et al., eds., 1994); and Gene Transfer and
Expression: A
Laboratory Manual (Kriegler, 1990).
Unless defined otherwise herein, all technical and scientific terms used
herein have the same
meaning as commonly understood by one of ordinary skill in the art to which
this invention
pertains.
Singleton, et al., Dictionary of Microbiology and Molecular Biology, 2nd ed.,
John Wiley and
Sons, New York (1994), and Hale & Markham, The Harper Collins Dictionary of
Biology,
Harper Perennial, New York (1991) provide one of skill in the art with a
general dictionary of
many of the biotechnology related terms used in this invention. Any methods
and materials
similar or equivalent to those described herein can be used in the practice or
testing of the
present invention.
Numeric ranges provided herein are inclusive of the numbers defining the
range.
Unless otherwise indicated, nucleic acids are written left to right in 5' to
3' orientation; amino
acid sequences are written left to right in amino to carboxy orientation,
respectively.
The following terms and phrases are defined for clarity:
As used here, the term "adjusting" means the process of treating a textile
material for a
sufficient length of time and under appropriate pH and temperature conditions
to produce a
lighter colour in said textile material by removal, modification or masking of
colour-causing
compounds in the textile material. Thus, "adjusting" refers to the treatment
of a textile material
to effect a brightening of the textile material.
In some embodiments, brightening is defined as a percentage of colour removal
from the
textile material. The amount of colour removal can be determined by comparing
the colour
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level can be determined by comparing the colour level following treatment with
an enzymatic
textile treatment composition according to the invention (i.e. residual colour
level) to the colour
level of the starting textile material (i.e. original colour level) using
known spectrophotometric
or visual inspection methods.
As used here, the term "original colour level" refers to the colour level of a
dyed textile material
prior to contact with an enzymatic textile treatment composition according to
the invention.
Original colour level may be measured using known spectrophotometric or visual
inspection
methods.
As used here, the term "residual colour level" refers to the colour level of a
dyed textile material
after contact with an enzymatic textile treatment composition according to the
invention.
Residual colour level may be measured using known spectrophotometric or visual
inspection
methods.
As used here, the term "cellulosic textile fibre material" refers to a
material that comprises
natural cellulosic fibres such as cotton, linen and hemp, semi-synthetic
cellulosic fibres such as
viscose and lyocell as well as blends of cellulosic fibres and synthetic
fibres such as elastane.
Suitable cellulosic textile fibre material substrates that can be treated with
the method
according to the invention are yarns, wovens, knits and garments.
As used here, a "perhydrolase" refers to an enzyme that is capable of
catalyzing a
perhydrolysis reaction that results in the production of a sufficiently high
amount of peracid
suitable for use in an enzymatic textile adjusting composition according to
the method
described herein. Generally, a perhydrolase enzyme used in the methods
described herein
exhibits a high perhydrolysis to hydrolysis ratio. In some embodiments, the
perhydrolase
comprises, consists of, or consists essentially of the Mycobacterium smegmatis
perhydrolase
amino acid sequence set forth in SEQ ID NO: 1, or a variant or homolog
thereof. In some
embodiments, the perhydrolase enzyme comprises acyl transferase activity and
catalyzes an
aqueous acyl transfer reaction.
As used here, a "peracid" is an organic acid of the formula RC(=O)OOH, wherein
R is an
aliphatic, aromatic or araliphatic radical.
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As used here, an "ester substrate" is a perhydrolase substrate that contains
an ester linkage.
Esters comprising aliphatic and/or aromatic carboxylic acids and alcohols may
be utilized as
substrates with perhydrolase enzymes. In some embodiments, the ester source is
selected
from the esters of one or more of the following acids: formic acid, acetic
acid, propionic acid,
butyric acid, valeric acid, caproic acid, caprylic acid, nonanoic acid,
decanoic acid, dodecanoic
acid, myristic acid, palmitic acid, stearic acid, and oleic acid. In some
embodiments, the ester
source is an acetate ester. In some embodiments, the ester source is selected
from one or
more of propylene glycol diacetate, ethylene glycol diacetate, glycerol
triacetate, ethyl acetate,
and glycerol tributyrate.
As used here, the terms "perhydrolyzation," "perhydrolyze" and "perhydrolysis"
refer to a
reaction wherein a peracid is generated from an ester substrate and a hydrogen
peroxide
source. The perhydrolyzation reaction is catalyzed with a perhydrolase, e.g.,
acyl transferase
or aryl esterase, enzyme. In some embodiments, a peracid is produced by
perhydrolysis of an
ester substrate of the formula RC(=O)OR*, where R and R* are the same or
different organic
moieties, in the presence of hydrogen peroxide (H202). In one embodiment, -OR*
is -OH. In
one embodiment, -OR* is replaced by -NH2. In some embodiments, a peracid is
produced by
perhydrolysis of a carboxylic acid or amide substrate.
As used here, the term "peracid," as used herein, refers to a molecule derived
from, or capable
of being derived from, a carboxylic acid ester which has been reacted with
hydrogen peroxide
to form a highly reactive product that is able to transfer one of its oxygen
atoms. It is this ability
to transfer oxygen atoms that enables a peracid, for example, peracetic acid,
to function as a
brightening agent.
As used here, the phrase "perhydrolysis to hydrolysis ratio" refers to the
ratio of the amount of
enzymatically produced peracid to the amount of enzymatically produced acid by
a
perhydrolase enzyme from an ester substrate under defined conditions and
within a defined
time. In some embodiments, the assays provided in WO 05/056782 are used to
determine the
amounts of peracid and acid produced by the enzyme.
As used herein, "effective amount of perhydrolase enzyme" refers to the
quantity of
perhydrolase enzyme necessary to achieve the enzymatic activity required in
the processes or
methods described herein. Such effective amounts are readily ascertained by
one of ordinary
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skill in the art and are based on many factors, such as the particular enzyme
variant used, the
pH used, the temperature used and the like, as well as the results desired
(e.g., level of
brightening).
As used herein, the term "transferase" refers to an enzyme that catalyzes the
transfer of a
functional group from one substrate to another substrate. For example, an acyl
transferase
may transfer an acyl group from an ester substrate to a hydrogen peroxide
substrate to form a
peracid.
As used herein, the term "acyl" refers to an organic group with the general
formula RCO-,
derived from an organic acid by removal of the -OH group. Typically, acyl
group names end
with the suffix "-oyl," e.g., ethanoyl chloride, CH3CO-CI, is the acyl
chloride formed from
ethanoic acid, CH3CO-OH.
As used herein, the term "acylation" refers to a chemical transformation in
which one of the
substituents of a molecule is substituted by an acyl group, or the process of
introduction of an
acyl group into a molecule.
As used herein, an "oxidizing chemical" is a chemical capable of brightening a
textile. The
oxidizing chemical is present at an amount, pH and temperature suitable for
brightening. The
term includes, but is not limited to hydrogen peroxide and peracids.
As used herein, the terms "purified" and "isolated" refer to the removal of
contaminants from a
sample and/or to a material (e.g., a protein, nucleic acid, cell, etc.), such
that the sample or
material is removed from at least one component with which it is naturally
associated. For
example, these terms may refer to a material which is substantially or
essentially free from
components which normally accompany it as found in its native state, such as,
for example, an
intact biological system.
As used herein, the term "polynucleotide" refers to a polymeric form of
nucleotides of any
length and any three-dimensional structure and single- or multi-stranded
(e.g., single-stranded,
double-stranded, triple-helical, etc.), which contain deoxyribonucleotides,
ribonucleotides,
and/or analogs or modified forms of deoxyribonucleotides or ribonucleotides,
including
modified nucleotides or bases or their analogs. Because the genetic code is
degenerate, more
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than one codon may be used to encode a particular amino acid, and the
polynucleotides
applied applied within the context of the present invention encode a
particular amino acid
sequence. Any type of modified nucleotide or nucleotide analog may be used, so
long as the
polynucleotide retains the desired functionality under conditions of use,
including modifications
that increase nuclease resistance (e.g., deoxy, 2'-O--Me, phosphorothioates,
etc.). Labels
may also be incorporated for purposes of detection or capture, for example,
radioactive or
nonradioactive labels or anchors, e.g., biotin. The term polynucleotide also
includes peptide
nucleic acids (PNA). Polynucleotides may be naturally occurring or non-
naturally occurring.
The terms "polynucleotide" and "nucleic acid" and "oligonucleotide" are used
herein
interchangeably. Polynucleotides of the invention may contain RNA, DNA, or
both, and/or
modified forms and/or analogs thereof. A sequence of nucleotides may be
interrupted by
non-nucleotide components. One or more phosphodiester linkages may be replaced
by
alternative linking groups. These alternative linking groups include, but are
not limited to,
embodiments wherein phosphate is replaced by P(O)S ("thioate"), P(S)S
("dithioate"), (O)NR2
("amidate"), P(O)R, P(O)OR', CO or CH2 ("formacetal"), in which each R or R'
is independently
H or substituted or unsubstituted alkyl (C,-C2o) optionally containing an
ether (-0-) linkage, aryl,
alkenyl, cycloalkyl, cycloalkenyl or araldyl. Not all linkages in a
polynucleotide need be
identical. Polynucleotides may be linear or circular or comprise a combination
of linear and
circular portions. Suitable polynucleotides are described in WO 2005/056782.
As used herein, the term "polypeptide" refers to any composition comprised of
amino acids and
recognized as a protein by those of skill in the art. The conventional one-
letter or three-letter
code for amino acid residues is used herein. The terms "polypeptide" and
"protein" are used
interchangeably herein to refer to polymers of amino acids of any length. The
polymer may be
linear or branched, it may comprise modified amino acids, and it may be
interrupted by
non-amino acids. The terms also encompass an amino acid polymer that has been
modified
naturally or by intervention; for example, disulfide bond formation,
glycosylation, lipidation,
acetylation, phosphorylation, or any other manipulation or modification, such
as conjugation
with a labeling component. Also included within the definition are, for
example, polypeptides
containing one or more analogs of an amino acid (including, for example,
unnatural amino
acids, etc.), as well as other modifications known in the art.
As used herein, the terms "analogous sequence," "homologous protein," "wild-
type or native
proteins," "wild-type sequence," "native sequence," "naturally-occuring
sequence," "wild-type
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gene," "related proteins," "derivative proteins," "variant proteins" and the
like, are familiar to
those skilled in the art and are described in more detail in WO 2005/056782 on
pages 12, 13
and 50 to 52, which are herein incorporated by reference. In some embodiments,
homologous
proteins are engineered to produce enzymes with desired activity(ies).
Several methods are known in the art that are suitable for generating variants
of the enzymes
described herein, including but not limited to site-saturation mutagenesis,
scanning
mutagenesis, insertional mutagenesis, random mutagenesis, site-directed
mutagenesis, and
directed-evolution, as well as various other recombinatorial approaches.
The degree of homology between sequences may be determined using any suitable
method
known in the art. For example, PILEUP is a useful program to determine
sequence homology
levels. PILEUP creates a multiple sequence alignment from a group of related
sequences
using progressive, pairwise alignments. It can also plot a tree showing the
clustering
relationships used to create the alignment. Another example of a useful
algorithm is the BLAST
algorithm. Useful methods and programs are referred to in WO 2005/056782 on
pages 59 and
60, which are herein incorporated by reference.
The terms "substantially similar" and "substantially identical" as generally
used in the context of
a polynucleotide or polypeptide sequence compared to a reference (i.e., wild-
type) sequence
as well as methods to determine sequence identity are described in more detail
in WO
2005/056782 on pages 61 and 62, which are herein incorporated by reference.
As used herein, the term "surfactant" refers to a substance that reduces
surface tension of a
liquid.
As used herein, the term "emulsifier" refers to a substance that promotes the
suspension of
one liquid in another.
As used herein, the term "sequestering agent" refers to a substance capable of
reacting with
metallic ions by forming a water-soluble complex in which the metal is held in
a non-ionizable
form.
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As used herein, the term "catalase" refers to an enzyme (i.e., a polypeptide
having catalytic
activity) that catalyzes the decomposition of hydrogen peroxide to water and
oxygen.
As used herein, the term the term "batch process" or "batchwise process" or
"discontinuous
process" or "exhaust process" refers to processing of textiles as lots or
batches in which the
whole of each batch is subjected to a process or one stage of a process at a
time.
As used herein, the term "liquor ratio" refers to the ratio of the weight of
liquor (liquid) employed
in a textile treatment process to the weight of the textile treated.
The enzymatic colour tone adjusting composition used in accordance with the
enzymatic
textile adjusting method of the present invention contain a perhydrolase
enzyme, an ester
substrate for the perhydrolase enzyme suitable for production of a peracid
upon catalytic
reaction of the perhydrolase enzyme on the substrate in the presence of a
hydrogen peroxide
source and/or hydrogen peroxide. The enzymatic colour tone adjusting
composition may,
optionally, further contain a surfactant and/or an emulsifier, a peroxide
stabilizer, a
fluorescence whitening agent, an enzymatic desizing agent, a biopolishing
agent, a
combination product, a sequestering agent or a bufferwhich maintains a pH of
about 6 to about
8 during a textile colour tone adjusting process.
In the following the components applied in accordance with the method of the
present
invention along with information on the quantities of these components are
described in more
detail. Parts (ppm) are parts by weight, unless noted otherwise.
Perhydrolase Enzyme
One or more perhydrolase enzymes may be used in the compositions according to
the
methods for enzymatic textile colour tone adjusting as described herein.
In some embodiments, the perhydrolase enzyme is naturally-occurring (i.e., a
perhydrolase
enzyme encoded by a genome of a cell). In some embodiments, the perhydrolase
enzyme
comprises, consists of, or consists essentially of an amino acid sequence that
is at least about
80%, 85%, 90%, 95%, 97%, 98%, 99%, or 99.5% identical to the amino acid
sequence of a
naturally-occurring perhydrolase enzyme.
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In some embodiments, the perhydrolase enzyme is a naturally-occurring M.
smegmatis
perhydrolase enzyme. In some embodiments, the perhydrolase enzyme comprises,
consists
of, or consists essentially of the amino acid sequence set forth in SEQ ID
NO:1 or a variant or
homologue thereof. In some embodiments, the perhydrolase enzyme comprises,
consists of,
or consists essentially of an amino acid sequence that is at least about 80%,
85%, 90%, 95%,
97%, 98%, 99%, or 99.5% identical to the amino acid sequence set forth in SEQ
ID NO:1.
The amino acid sequence of M. smegmatis perhydrolase is shown below:
MAKRI LCFGDSLTWGWVPVEDGAPTERFAPDVRWTGVLAQQLGADFEVI EEGLSARTTN I D
DPTDPRLNGASYLPSCLATHLPLDLVIIMLGTNDTKAYFRRTPLDIALGMSVLVTQVLTSAGG
VGTTYPAPKVLVVSPPPLAPMPHPWFQLIFEGGEQKTTELARVYSALASFMKVPFFDAGSVI
STDGVDGIHFTEANNRDLGVALAEQVRSLL (SEQ ID NO:1).
The corresponding polynucleotide sequence encoding M. smegmatis perhydrolase
is:
5'-ATGGCCAAGCGAATTCTGTGTTTCGGTGATTCCCTGACCTGGGGCTGGGTCC
CCGTCGAAGACGGGGCACCCACCGAGCGGTTCGCCCCCGACGTGCGCTGGACCGGTG
TGCTGGCCCAGCAGCTCGGAGCGGACTTCGAGGTGATCGAGGAGGGACTGAGCGCGC
GCACCACCAACATCGACGACCCCACCGATCCGCGGCTCAACGGCGCGAGCTACCTGC
CGTCGTGCCTCGCGACGCACCTGCCGCTCGACCTGGTGATCATCATGCTGGGCACCAA
CGACACCAAGGCCTACTTCCGGCGCACCCCGCTCGACATCGCGCTGGGCATGTCGGT
GCTCGTCACGCAGGTGCTCACCAGCGCGGGCGGCGTCGGCACCACGTACCCGGCACC
CAAGGTGCTGGTGGTCTCGCCGCCACCGCTGGCGCCCATGCCGCACCCCTGGTTCCA
GTTGATCTTCGAGGGCGGCGAGCAGAAGACCACTGAGCTCGCCCGCGTGTACAGCGC
GCTCGCGTCGTTCATGAAGGTGCCGTTCTTCGACGCGGGTTCGGTGATCAGCACCGAC
GGCGTCGACGGAATCCACTTCACCGAGGCCAACAATCGCGATCTCGGGGTGGCCCTC
GCGGAACAGGTGCGGAGCCTGCTGTAA-3' (SEQ ID NO:2).
In some embodiments, the perhydrolase enzyme comprises one or more
substitutions at one
or more amino acid positions equivalent to position(s) in the M. smegmatis
perhydrolase amino
acid sequence set forth in SEQ ID NO:1. In some embodiments, the perhydrolase
enzyme
comprises any one or any combination of substitutions of amino acids selected
from M1, K3,
R4,15, L6, C7, D10, S11, L12, T13, W14, W16, G15, V17, P18, V19, D21, G22,
A23, P24, T25,
E26, R27, F28, A29, P30, D31, V32, R33, W34, T35, G36, L38, Q40, Q41, D45,
L42, G43, A44,
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F46, E47, V48, 149, E50, E51, G52, L53, S54, A55, R56, T57, T58, N59, 160,
D61, D62, P63,
T64, D65, P66, R67, L68, N69, G70, A71, S72, Y73, S76, C77, L78, A79, T80,
L82, P83, L84,
D85, L86, V87, N94, D95, T96, K97, Y99F100, R101, R102, P104, L105, D106,
1107, A108,
L109, G110, M111, S112, V113, L114, V115, T116, Q117, V118, L119, T120, S121,
A122,
G124, V125, G126, T127, T128, Y129, P146, P148, W149, F150, 1153, F154, 1194,
and F196.
In some embodiments, the perhydrolase enzyme comprises one or more of the
following
substitutions at one or more amino acid positions equivalent to position(s) in
the M. smegmatis
perhydrolase amino acid sequence set forth in SEQ ID NO:1: L1 2C, Q, or G;
T25S, G, or P;
L53H, Q, G, or S; S54V, L A, P, T, or R; A55G or T; R67T, Q, N, G, E, L, or F;
K97R; V125S, G,
R, A, or P; F154Y; F196G.
In some embodiments, the perhydrolase enzyme is the S54V variant of SEQ ID NO:
1, which is
shown, below as SEQ ID NO: 3, wherein the S54V substitution is underlined.
The amino acid sequence of M. smegmatis perhydrolase is shown below:
MAKRILCFGDSLTWGWVPVEDGAPTERFAPDVRWTGVLAQQLGADFEVIEEGLVARTTNI D
DPTDPRLNGASYLPSCLATHLPLDLVIIMLGTNDTKAYFRRTPLDIALGMSVLVTQVLTSAGG
VGTTYPAPKVLVVSPPPLAPMPHPWFQLIFEGGEQKTTELARVYSALASFMKVPFFDAGSVI
STDGVDGIHFTEANNRDLGVALAEQVRSLL (SEQ ID NO: 3).
In some embodiments, the perhydrolase enzyme comprises a combination of amino
acid
substitutions at amino acid positions equivalent to amino acid positions in
the M. smegmatis
perhydrolase amino acid sequence set forth in SEQ ID NO:1: L121 S54V; L12M
S54T; L12T
S54V; L12Q T25S S54V; L53H S54V; S54P V125R; S54V V125G; S54V F196G; S54V K97R
V125G; or A55G R67T K97R V1 25G.
In some embodiments, the perhydrolase enzyme comprises a perhydrolysis to
hydrolysis ratio
of at least 1. In some embodiments, the perhydrolase enzyme comprises a
perhydrolysis to
hydrolysis ratio greater than 1.
In some embodiments, the perhydrolase enzyme is provided in the enzymatic
textile colour
tone adjusting composition used according to the textile colour tone adjusting
method of the
present invention at a concentration of about 0.5 to about 2.5 ppm, about 1.5
to about 2.0 ppm,
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for example, about 1.7 ppm, based on the total weight of the aqueous
composition (bath)
applied for treatment of the textile material.
Ester Substrate
The enzymatic colour tone adjusting compositions used in accordance with the
method
described herein include an ester which serves as a substrate for the
perhydrolase enzyme for
production of a peracid in the presence of hydrogen peroxide. In some
embodiments, the ester
substrate is an ester of an aliphatic and/or aromatic carboxylic acid. In some
embodiments,
the ester substrate is an ester of one or more of the following: formic acid,
acetic acid, propionic
acid, butyric acid, valeric acid, caproic acid, caprylic acid, nonanoic acid,
decanoic acid,
dodecanoic acid, myristic acid, palmitic acid, stearic acid, and oleic acid.
In some
embodiments, glycerol triacetate, glycerol tributyrate, and other esters serve
as acyl donors for
peracid formation. In some embodiments, the ester substrate is selected from
propylene glycol
diacetate, ethylene glycol diacetate, glycerol triacetate, ethyl acetate, and
glycerol tributyrate.
In some embodiments, the ester substrate is propylene glycol diacetate,
ethylene glycol
diacetate, or ethyl acetate. In one embodiment, the ester substrate is
propylene glycol
diacetate.
In some embodiments, the ester substrate, for example, propylene glycol
diacetate, is
provided at a concentration of about 2000 to about 4000 ppm, about 2500 to
about 3500 ppm,
about 2800 ppm to about 3200 ppm, or about 3000 ppm, based on the total weight
of the
aqueous composition (bath) applied for treatment of the textile material.
Hydrogen Peroxide Source
The enzymatic colour tone adjusting compositions used in accordance with the
method
described herein include a hydrogen peroxide source. Hydrogen peroxide can be
either added
directly in batch, or generated continuously "in situ" by chemical, electro-
chemical, and/or
enzymatic means.
In some embodiments, the hydrogen peroxide source is hydrogen peroxide. In
some
embodiments, the hydrogen peroxide source is a solid compound that generates
hydrogen
peroxide spontaneously upon addition to water. Such compounds include adducts
of
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hydrogen peroxide with various inorganic or organic compounds, of which the
most widely
employed is sodium carbonate perhydrate, also referred to as sodium
percarbonate.
Inorganic perhydrate salts are one embodiment of hydrogen peroxide source.
Examples of
inorganic perhydrate salts include perborate, percarbonate, perphosphate,
persulfate and
persilicate salts. The inorganic perhydrate salts are normally the alkali
metal salts.
Other hydrogen peroxide adducts useful in the compositions used in accordance
with the
method described herein include adducts of hydrogen peroxide with zeolites, or
urea hydrogen
peroxide.
The hydrogen peroxide source compounds may be included as the crystalline
and/or
substantially pure solid without additional protection. For certain perhydrate
salts however, the
preferred executions of such granular compositions utilize a coated form of
the material which
provides better storage stability for the perhydrate salt in the granular
product. Suitable
coatings comprise inorganic salts such as alkali metal silicate, carbonate or
borate salts or
mixtures thereof, or organic materials such as waxes, oils, or fatty soaps.
In some embodiments, the hydrogen peroxide source is an enzymatic hydrogen
peroxide
generation system. In one embodiment, the enzymatic hydrogen peroxide
generation system
comprises an oxidase and its substrate. Suitable oxidase enzymes include, but
are not limited
to: glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase,
alcohol oxidase,
glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyalcohol
oxidase, L-amino
acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine
oxidase, lysine
oxidase, lactate oxidase, vanillyl oxidase, glycolate oxidase, galactose
oxidase, uricase,
oxalate oxidase, and xanthine oxidase.
The following equation provides an example of a coupled system for enzymatic
production of
hydrogen peroxide.
Glucose oxidase
Glucose + H2O ---------------------------------------------------gluconic acid
+ H202
Perhydrolase
H202 + ester substrate ----------------------------------------4 alcohol +
peracid
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It is not intended that the present invention be limited to any specific
enzyme, as any enzyme
that generates H202 with a suitable substrate may be used in the present
invention. For
example, lactate oxidases from Lactobacillus species which are known to create
H202 from
lactic acid and oxygen may be used. One advantage of the enzymatic generation
of acid (e.g.,
gluconic acid in the above example) is that this reduces the pH of a basic
solution to the pH
range in which a peracid is most effective in colour tone adjusting (i.e., at
or below the pKa).
Other enzymes (e.g., alcohol oxidase, ethylene glycol oxidase, glycerol
oxidase, amino acid
oxidase, etc.) that can generate hydrogen peroxide also may be used with ester
substrates in
combination with the perhydrolase enzymes of the present invention to generate
peracids.
In some embodiments, the hydrogen peroxide generating oxidase is a
carbohydrate oxidase.
Hydrogen peroxide may also be generated electrochemically, for example using a
fuel cell fed
oxygen and hydrogen gas.
In some embodiments, the hydrogen peroxide source is hydrogen peroxide
provided at a
concentration of about 1000 to about 3200 ppm, about 1500 to about 2800 ppm,
about 2000
ppm to about 2200 ppm, or about 2100 ppm, based on the total weight of the
aqueous
composition (bath) applied for treatment of the textile material.
Surfactants and Emulsifier
The enzymatic textile colour tone adjusting compositions used in accordance
with the present
method may contain one or more, i.e., at least one surfactant and/or at least
one emulsifier.
Surfactants suitable for use in practicing the present invention include,
without limitation,
nonionic (see, e.g., U.S. Pat. No. 4,565,647, which is herein incorporated by
reference);
anionic; cationic; and zwitterionic surfactants (see, e.g., U.S. Pat. No.
3,929,678 which is
herein incorporated by reference). Anionic surfactants include, without
limitation, linear
alkylbenzenesulfonate, a-olefinsulfonate, alkyl sulfate (fatty alcohol
sulfate), alcohol
ethoxysulfate, secondary alkanesulfonate, alpha-sulfo fatty acid methyl ester,
alkyl- or
alkenylsuccinic acid, and soap. Non-ionic surfactants include, without
limitation, fatty alcohol
ethoxylate, isotridecanol ethoxylate, nonylphenol ethoxylate,
alkylpolyglycoside,
alkyldimethylamineoxide, ethoxylated fatty acid monoethanolamide, fatty acid
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monoethanolamide, polyhydroxy alkyl fatty acid amide, and N-acyl N-alkyl
derivatives of
glucosamine ("glucamides").
In some embodiments, the surfactant and/or emulsifier comprises a non-ionic
surfactant. In
one embodiment, the non-ionic surfactant is a fatty alcohol ethoxylate. In one
embodiment, the
non-ionic surfactant is isotridecanol ethoxylate. In one embodiment, the non-
ionic surfactant is
a fatty alcohol ethoxylate and isotridecanol ethoxylate.
In one embodiment, the composition used in accordance with the present method
comprises a
surfactant and an emulsifier.
A surfactant may be present at a concentration of about 300 ppm to about 4800
ppm, about
600 ppm to about 3600 ppm, or about 300 ppm to about 1200 ppm, based on the
total weight of
the aqueous composition (bath) applied for treatment of the textile material.
In some embodiments, the enzymatic colour tone adjusting composition contains
isotridecanol
ethoxylate at a concentration of about 300 ppm to about 3600 ppm, about 600
ppm to about
3000 ppm, or about 900 ppm to about 2400 ppm, based on the total weight of the
aqueous
composition (bath) applied for treatment of the textile material.
Peroxide Stabilizer
The enzymatic colour tone adjusting compositions used in accordance with the
method
described herein may contain a peroxide stabilizer. Examples of peroxide
stabilizers include,
but are not limited to, sodium silicate, sodium carbonate, acrylic polymers,
magnesium salts,
and phosphonic acid. In one embodiment, the peroxide stabilizer is phosphonic
acid.
A peroxide stabilizer may be present in the enzymatic textile colour tone
adjusting composition
at a concentration of about 60 ppm to about 600 ppm, about 60 ppm to about
1200 ppm, or
about 120 ppm to about 960 ppm based on the total weight of the aqueous
composition (bath)
applied for treatment of the textile material.
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Combination Product
Appropriately, at least one surfactant and/or emulsifier, at least one
peroxide stabilizer and at
least one sequestering agent are applied as a combination product containing
each of at least
one surfactant and/or emulsifier, at least one peroxide stabilizer and at
least one sequestering
agent. The said combination product is designated bleaching processor and is
commercially
available, for example, CLARITE LTC, CLARITE WIN or CLARITE ONE (products
of
Huntsman).
A surfactant may be present at a concentration of about 5% to about 40%, about
20% to about
30%, or about 5% to about 10%, based on the total weight of the bleaching
processor.
A peroxide stabilizer may be present in the combination product at a
concentration of about 1 %
to about 5%, about 1 % to about 10%, or about 2% to about 8%, based on the
total weight of the
bleaching processor.
A sequestering agent may be present in the bleaching processor at a
concentration of about
1 % to about 15%, about 5% to about 10%, or about 3% to about 10%, based on
the total
weight of the bleaching processor.
In some embodiments, the bleaching processor contains isotridecanol ethoxylate
at a
concentration of about 5% to about 30%, about 10% to about 25%, or about 15%
to about 20%,
based on the total weight of the bleaching processor.
The bleaching processor is suitably provided as an aqueous composition
comprising the
above indicated components.
Buffer
The enzymatic colour tone adjusting composition may contain a buffer that is
capable of
maintaining the pH of the composition at a pH of about 6 to about 8. The
buffer may be, for
example, a phosphate buffer, pH 7, or a sodium carbonate or potassium
carbonate, pH 7.
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Enzymatic Textile Colour Tone Adjusting Method
The method according to the invention is particularly suitable for the
treatment of
denim dyed with vat dyes, reactive dyes, direct dyes and sulphur dyes, most
preferably
for indigo-dyed denim and/or sulfur-dyed denim. The method provides textile
material
with a soft handle and very good crease recovery properties compared to
textiles
treated using conventional methods. The methods described herein are
particularly
carried out as a discontinuous process but can also be carried out as a
semi-continuous process like pad-batch or pad-roll.
Appropriately, the method of the invention utilizes a liquor ratio of about
2:1 to about 50:1,
about 5:1 to about 20:1, for example, about 20:1 or 10:1.
Textiles are contacted with the enzymatic colour tone adjusting composition at
a temperature
of about 55 C to about 75 C, about 60 C to about 70 C, for a processing time
of about 20 to
about 60 minutes at a pH of about 6 to about 8. In one embodiment, the
treatment temperature
is about 65 C and the processing time is about 50 minutes. In some
embodiments, the
temperature of the enzymatic colour tone adjusting composition is raised by
about 2 C per
minute from a starting temperature of about 20 C to about 50 C, for example,
about 20 C to
about 40 C, until the processing temperature for colour tone adjusting is
reached.
One or more rinsing steps are performed after treatment of the textile
material with the
enzymatic colour tone adjusting composition, to remove the colour tone
adjusting composition.
Appropriately, the textile is rinsed with an aqueous composition (water or a
composition
containing water). In some embodiments, the rinsing temperature is about 40 C
to about 60 C,
for example, about 50 C.
In some embodiments, the aqueous rinsing composition contains a catalase
enzyme to
catalyze the decomposition of hydrogen peroxide to water and oxygen. In one
embodiment,
the textile is rinsed twice with a catalase containing aqueous composition for
about 10 minutes
for each rinse. In one embodiment residual hydrogen peroxide is removed by
rinsing twice with
an aqueous composition containing catalase at about 50 C. In some embodiments,
catalase
may be added directly to the liquor containing the perhydrolase enzyme without
first dropping
the bath.
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As the enzymes that hydrolyze polyester substrates suitable for use in the
present invention
may be used, for example, pectinases, cutinases or lipases. These enzymes,
their application
as well as assays to determine enzyme activity are described and referred to
in more detail in
WO 2007/136469 on pages 21 and 22, which are herein incorporated by reference.
Further suitable enzymes that may be used additionally in the method according
to the
invention are amylases (desizing agents) and cellulases (biopolishing agents).
The following examples are intended to illustrate, but not limit, the
invention. Temperatures are
in degrees Celsius, parts are parts by weight and the percentage data are
percentages by
weight, unless noted otherwise. Parts by weight bear the same relation to
parts by volume as
the kilogram to the litre.
Experimental Part
A comparison between the method of the present invention and conventional
oxidation bleach
with hydrogen peroxide is performed according to the procedures given below by
treating the
fabric in exhaust using a Mathis AG Labomat.
Examples 1 and 2 and Comparative Example 3
Enzymatic colour tone adjusting (Examples 1 and 2) carried out as a one-bath
exhaustion
method:
Indigo dyed denim swatches (length: 11 cm, width: 9.5 cm), washed at 65 C/5
min and cold in
overflow/5 min, are treated in a bath containing the combination product, the
buffer, propylene
glycol diacetate, hydrogen peroxide and the perhydrolase enzyme in the amounts
given in
Table 1 using a liquor ratio of 10:1. The temperature is raised from ambient
temperature to a
target temperature of 65 C at a rate of 2 C per minute. The bath is then held
at 65 C for 5 0
minutes and after cooling and draining the swatches are rinsed twice for 10
minutes each at
50 C and then dried at 70 C. 0.5 g/I of a 25% solution of Catalase T100
(available from
Genencor) is included in each rinse.
Alkali colour tone adjusting (Comparative Example 3) carried out as a one-bath
exhaustion
method:
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The procedure described above is repeated, but perhydrolase enzyme and
phosphate buffer
are replaced with sodium hydroxide.
Table 1: Examples 1, 2 and Comparative Example 3
Example # 1 2 3 (comparison)
Deionised water [g/I] 975.0 975.0 987.5
INVALON EC 1) [g/I] 1.0
CLARITE GS 3) [g/I] 0.5
CLARITE WIN 2) [g/I] 3.0
CLARITE ONE 4) [g/I] 3.0
Phosphate buffer solution [ml/1] 12.0 12.0
Propylene glycol diacetate [ml/1] 3.0 3.0
Hydrogen peroxide 35% [ml/1] 6.0 6.0 6.0
Perhydrolase enzyme 5) [g/I1 1.0 1.0
NaOH (30 %) [g/I] 5.0
pH 10.2 10.5 11.0
Aspect (initially) Colourless, dull Colourless, dull Colourless, dull
pH after treatment 7.2 7.2 11.5
Aspect (after treatment) Dark green, Dark green, Dark green, clear
clear clear
INVALON EC: dispersing agent (commercial product supplied by Huntsman)
2) CLARITE WIN: bleaching processor (commercial product supplied by Huntsman)
3) CLARITE GS: bleaching processor (commercial product supplied by Huntsman)
4) CLARITE ONE: bleaching processor (commercial product supplied by Huntsman)
5) Primagreen EcoWhite (1x) (commercial product supplied by Genencor)
Example 4
Enzymatic on-tone washdown carried out as a one-bath exhaustion method:
Indigo dyed denim swatches (length: 11 cm, width: 9.5 cm), desized at 10:1
liquor ratio, 60
C/10 min, with 0.5 g/I CLARITE WIN, 0.5 g/I Albfluid C and 1.5 g/I ULTRAVON
RW
(non-ionic surfactant, commercial product supplied by Huntsman) are treated in
the same bath
with 1.5 g/I of INVAZYME LTE (perhydrolase enzyme) at 10:1 liquor ratio, 60
C/10 min.
Following desizing, the denim is stonewashed in a rotary washing machine with
1 kg of pumice
stones (60 C/40 min). Subsequently, 1.0 g/I CLARITE LTC (combination
product, supplied
by Huntsman), 2.5 g/I of soda ash, 3.0 g/I of propylene glycol diacetate, 3.0
g/I of hydrogen
peroxide and 1.0 g/I of INVAZYME LTE (perhydrolase enzyme) are added to the
liquor. The
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bath is then held at 65 C for 50 minutes and after cooling and draining the
swatches are rinsed
twice for 15 minutes each at 50 C with a bath containing 0.5 g/I INVAZYME
CAT (stabilized
liquid catalase enzyme to remove residual peroxide) and afterwards for 15 min
with a bath
containing 1.0 g/I of a softening agent (Turpex CAN new) at room temperature,
and then dried.
The denim swatches thus obtained have a soft handle and very good crease
recovery
properties.
Example 5
Effect of Perhydrolase Concentration on Dye Discoloration on Indigo-dyed Denim
Materials
Perhydrolase (PrimaGreen EcoWhite 1 (321 U/g), available from Genencor
Division,
Danisco US, Inc.), is used in this experiment. H202 analysis grade (30 wt %)
and propylene
glycol diacetate >99.7% (PDGA) were purchased from Sigma Aldrich.
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
in a Unimac
UF 50 washing machine under the following conditions:
= Desizing for 15 minutes at 10:1 liquor ratio 50 C with 0.5 g/I (15 g) of
Optisize 160
amylase (Genencor) and 0.5 g/I (15 g) of a non-ionic surfactant (Ultravon RW
(Huntsman)).
= 2 cold rinse steps for 5 minutes at 30:1 liquor ratio.
Following desizing, the denim is stonewashed in a Unimac UF 50 rotary washing
machine
according to the following program:
= Cold rinsing for 5 minutes at 10:1 liquor ratio
= Stonewashing for 60 minutes at 10:1 liquor ratio 55 C with 1 kg of pumice
stone, pH 6.5
- 7 (1 g/I of disodium phosphate 2H20 + 0.53 g/I of citric acid H2O) and 0.025
g/I of
MEX-500 neutral cellulase (Meiji).
= 2 cold rinse steps of 5 min each
The denim is dried in a household dryer and then used to make swatches (7 x 7
cm).
After stonewashing, the experiments are performed in a Launder-O-meter (Rapid
Laboratory
Dyeing Machine type H12) according to the following process:
= 450 ml stainless steel reaction vessels are filled with 100 ml of pH 8
phosphate buffer
(8.9 g/I of disodium phosphate 2H20 + 0.4 g/I of monosodium phosphate
anhydrous).
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= To each vessel five (7 x 7 cm) stonewashed denim swatches of 10 g weight are
added.
= 6 ml/I of H202 solution (30% wt) and 2m1/I of PDGA (>99.7%) is added.
= Perhydrolase is added at concentrations of 0. 01, 0. 05, 0. 3, 1.0, 3.0, or
10 ml/l.
= The reaction vessels are closed and loaded into the launder-O-meter, which
is
pre-heated to 60 C.
= Incubation is performed for 60 minutes, after which the swatches are rinsed
by
overflow, spun dry in an AEG IPX4 centrifuge, and dried with an Elna Press
Electronic
iron at program cotton and evaluated.
Evaluation of denim swatches
The denim swatches are evaluated after perhydrolase treatment with a Minolta
Chromameter
CR 310 in the CIE Lab color space with a D 65 light source. Measurements are
done before
and after perhydrolase treatment and the results from five swatches are
averaged. The total
color difference (TCD) is calculated using the formula: TCD = J (AL)2 + (Aa)2
+ (Ab)2.
The results are shown in Table 2.
Table 2
Perhydrolase TCD L/ a/ b
concentration (ml/1)
Buffer 0.44 0.41/0.13/0.10
0.01 0.56 0.40/0.32/-0.23
0.05 1.46 1.10/0.31/-0.90
0.3 1.97 1.50/0.34/-1.23
1 2.11 1.37/0.51/-1.52
3 2.05 1.41/0.41/-1.43
1.49 1.19/0.42/-0.80
Example 6
Effect of H202 and PDGA Concentrations on Dye Discoloration Performance of
Perhydrolase
on Indigo-dyed Denim
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
and
stonewashed as described in Example 5. After stonewashing, the experiments are
performed
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in a Launder-O-meter (Rapid Laboratory Dyeing Machine type H12) according to
the following
process:
= 450 ml stainless steel reaction vessels are filled with 100 ml of pH 8
phosphate buffer
(8.9 g/I of disodium phosphate 2H20 + 0.4 g/I of monosodium phosphate
anhydrous).
= To each vessel five (7 x 7 cm) stonewashed denim swatches of 10 g weight are
added.
= H202 solution (30% wt) and PDGA (>99.7%) are added according to the
experimental
design as shown in Table 3
=
Table 3
[H202] (MI/1) [PGDA] (ml/1)
7.55 3.8
15 7.5
0.1 7.5
7.55 3.8
0.1 0.1
15 0.1
7.55 3.8
6.0 3.0
0 3.0
6.0 0
15 3.8
7.55 7.5
= 1.0 mI/I of perhydrolase is added (PrimaGreen EcoWhite 1 (321 U/g)).
= The reaction vessels are closed and loaded into the Launder-O-Meter which
was
pre-heated to 60 C
= Incubation is performed for 60 minutes, after which the swatches are rinsed
by
overflow, spun dry in an AEG IPX4 centrifuge, and dried with an Elna Press
Electronic
iron at program cotton, and evaluated.
Evaluation of denim swatches
The denim swatches are evaluated after perhydrolase treatment with a Minolta
Chromameter
CR 310 in the CIE Lab color space with a D 65 light source. Measurements are
done before
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and after perhydrolase treatment and the results from five swatches are
averaged. The total
color difference (TCD) is calculated using the formula: TCD = J (AL)2 + (4a)2
+ (4b)2.
The results are shown in Table 4.
Table 4
[H202] (ml/1) [PGDA] (ml/1) TCD L/ a/ b
7.55 3.8 2.33 1.03/0.36/-1.24
15 7.5 2.48 1.11/0.40/-1.37
0.1 7.5 1.09 0.57/0.02/0.00
7.55 3.8 2.31 1.04/0.45/-1.17
0.1 0.1 0.76 0.07/-0.04/-0.06
15 0.1 1.48 0.66/0.12/-0.49
6.0 3.0 2.55 1.50/0.24/-1.17
0 3.0 0.62 0.15/-0.06/0.22
6.0 0 0.80 -0.22/0.10/-0.15
15 3.8 2.17 0.62/0.43/-1.28
7.55 7.5 2.37 1.17/0.35/-1.19
Example 7
Effect of Time on Dye Discoloration Performance of Perhydrolase on Indigo-dyed
Denim
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
and
stonewashed as described in Example 5. After stonewashing, the experiments are
performed
in a Launder-O-meter (Rapid Laboratory Dyeing Machine type H12) according to
the following
process.
= 450 ml stainless steel reaction vessels are filled with 100 ml of pH 8
phosphate buffer
(8.9 g/I Disodium phosphate 2H20 + 0.4 g/I Monosodium phosphate anhydrous)
= To each vessel five (7 x 7 cm) stonewashed denim swatches of 10 g weight are
added.
= 6 mI/I of H202 solution (30% wt) and 0.2 mI/I of PDGA (>99.7%) are added.
= 1.0 g/I of perhydrolase is added (PrimaGreen EcoWhite 1 (321 U/g)).
= The reaction vessels are closed and loaded into the Launder-O-Meter, which
was
pre-heated to 60 C.
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Incubation is performed for 10, 20, 30, 40, 50, or 60 minutes, after which the
swatches
are rinsed by overflow, spun dry in an AEG IPX4 centrifuge, dried with an Elna
Press
Electronic iron at program cotton, evaluated.
Evaluation of denim swatches
The denim swatches are evaluated after perhydrolase treatment with a Minolta
Chromameter
CR 310 in the CIE Lab color space with a D 65 light source. Measurements are
done before
and after perhydrolase treatment and the results from five swatches are
averaged. The total
color difference (TCD) is calculated using the formula: TCD = J (AL)2 + (Aa)2
+ (Ab)2.
[01] The results are shown in Table 5.
Table 5
time TCD L/ a/ b
buffer 1.09 1.05/0.27/0.05
1.48 0.97/0.30/-1.08
2.17 1.51/0.45/-1.49
2.05 1.28/0.53/-1.51
2.24 1.57/0.44/-1.55
2.45 1.80/0.49/-1.59
2.62 1.99/0.46/-1.64
Example 8
Effect of Temperature on Dye Discoloration Performance of Perhydrolase on
Indigo-dyed
Denim
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
and
stonewashed as described in Example 5. After stonewashing, the experiments are
performed
in a Launder-O-meter (Rapid Laboratory Dyeing Machine type H12) according to
the following
process.
= 450 ml stainless steel reaction vessels are filled with 100 ml of pH 8
phosphate buffer
(8.9 g/I of disodium phosphate 2H20 + 0.4 g/I of monosodium phosphate
anhydrous).
= To each vessel five (7 x 7 cm) stonewashed denim swatches of 10 g weight are
added.
= 6 ml/I of H202 solution (30% wt) and 2m1/I of PDGA (>99.7%) is added.
= 1.0 mI/l of perhydrolase is added (PrimaGreen EcoWhite 1 (321 U/g)).
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= The reaction vessels are closed and loaded into the Launder-O-Meter, which
was
pre-heated to 30, 40, 50 or 60 C.
= Incubation is performed for 60 minutes, the swatches rinsed by overflow,
spun dry in an
AEG IPX4 centrifuge, dried with an Elna Press Electronic iron at program
cotton,
evaluated.
Evaluation of denim swatches
The denim swatches are evaluated after perhydrolase treatment with a Minolta
Chromameter
CR 310 in the CIE Lab color space with a D 65 light source. Measurements are
done before
and after perhydrolase treatment and the results from five swatches are
averaged. The total
color difference (TCD) is calculated using the formula: TCD = J (AL)2 + (Aa)2
+ (Ab)2.
The results are shown in Table 6.
Table 6
Temperature C TCD L/ a/ b
30 (only buffer) 0.93 0.91/0.07/0.16
30 1.36 1.20/0.28/-0.57
40 (only buffer) 0.78 0.77/0.11/-0.02
40 1.55 1.26/0.28/-0.86
50 (only buffer) 1.07 1.06/0.11/-0.02
50 2.02 1.63/0.32/-1.14
60 (only buffer) 0.9 0.86/0.24/-0.15
60 2.21 1.67/0.44/-1.38
Example 9
Color Adjustment Performance with a Cellulase + Perhydrolase Sequential
Process on
Indigo-dyed Denim in a Front Loading Washing Machine
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
in a Unimac
UF 50 washing machine under the following conditions:
= Desizing for 15 minutes at 10:1 liquor ratio 50 C with 0.5 g/I (15 g) of
Optisize 160
amylase (Genencor) and 0.5 g/I (15 g) of a non-ionic surfactant (Ultravon RW
(Huntsman))
= 2 cold rinses for 5 minutes at 30:1 liquor ratio
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Following desizing, the denim is stonewashed in a Unimac UF 50 rotary washing
machine
according to the following procedure:
= Cold rinse for 5 minutes at 10:1 liquor ratio
= Stonewashing for 60 minutes at 10:1 liquor ratio 55 C with 1 kg of pumice
stone, pH 4.8
(1 g/I of trisodium citrate 2 H2O + 0.87g/l of citric acid H2O) 1.17g/l of
Indiage 2XL
cellulase (Genencor)
= 2 cold rinse steps of 5 min each
= 4 legs taken out as a control
After stonewashing, treatment with perhydrolase is performed in a Unimac UF 50
washing
machine according to the following process:
= 60 minutes at 10:1 liquor ratio, with 1 g/I perhydrolase (PrimaGreen
EcoWhite 1 (321
U/g)), 6 g/I of H202 solution (30%wt) and 3g/I of PDGA(>99.7%) at pH 7 (1 g/I
of
disodium phosphate 2 H2O and 0.17 g/I of citric acid) and temperature of 60 C.
The pH
was kept at 7 by adding 4 M of sodium hydroxide solution
= 2 cold rinses for 5 minutes at 30:1 liquor ratio
= The denim is dried in a household dryer.
Evaluation of denim legs
Colour tone adjusting of denim legs is evaluated after treatment with a
Minolta Chromameter
CR 310 in the CIE Lab color space with a D 65 light source. For each denim
leg, 8
measurements are taken and the results of the 12 legs (96 measurements) were
averaged.
The results are shown in Table 7.
Table 7
trials L/a/b
Perhydrolase treatment 36.3/-0.29/-15.17
Example 10
Color Adjustment Performance of Cellulase + Laccase + Perhydrolase Sequential
Process on
Indigo-dyed Denim in a Front Loading Washing Machine
Procedure
Denim, 12 legs (ACG denim style 80270) weighing approximately 3 kg, is desized
in a Unimac
UF 50 washing machine under the following conditions:
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= Desizing for 15 minutes at 10:1 liquor ratio 50 C with 0.5 g/I (15 g) of
Optisize 160
amylase (Genencor) and 0.5 g/I (15 g) of a non-ionic surfactant (Ultravon RW
(Huntsman).
= 2 cold rinses for 5 minutes at 30:1 liquor ratio
Following desizing, the denim is stonewashed in a Unimac UF 50 rotary washing
machine
according to the following procedure:
= Cold rinse for 5 minutes at 10:1 liquor ratio
= Stonewashing for 60 minutes at 10:1 liquor ratio 55 C with 1 kg of pumice
stone, pH 4.8
(1 g/I of trisodium citrate 2 H2O + 0.87g/l of citric acid H2O) and 1.17g/l of
Indiage 2XL
(Genencor)
= 2 cold rinse steps of 5 min each
After stonewashing, laccase treatment is performed in a Unimac UF 50 washing
machine
according to the following process:
= 30 minutes at 10:1 liquor ratio, with 3 g/I of ready to use PrimaGreen
EcoFade LT 100
(Genencor) laccase and laccase mediator at pH 6 and temperature of 30 C
= 2 cold rinses for 5 minutes at 30:1 liquor ratio
= The denim is dried in a household dryer.
After brightening with laccase, treatment with perhydrolase is performed in a
Unimac UF 50
washing machine according to the following process:
= 60 minutes at 10:1 liquor ratio, with 1 g/I of perhydrolase (PrimaGreen
EcoWhite 1
(321 U/g)), 6 g/I of H202 solution (30%wt) and 3g/I of PDGA(>99.7%) at pH 8
(8.9 g/I
disodium phosphate 2H20 + 0.4 g/I monosodium phosphate anhydrous) and
temperature of 60 C
= 2 cold rinses for 5 minutes at 30:1 liquor ratio
The denim is dried in a household dryer
Evaluation of denim legs
Brightening of denim legs is evaluated after laccase treatment and after
perhydrolase
treatment with a Minolta Chromameter CR 310 in the CIE Lab color space with a
D 65 light
source. For each denim leg, 8 measurements are taken and the results of the 12
legs (96
measurements) are averaged.
The results are shown in Table 8.
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Table 8
Trials L/a/b
Laccase 40.5/-1.5/-12.1
Laccase + Perhydrolase 44.4/-1.3/-15.2
Example 11
Effect of Perhydrolase Treatment on a Dyed Fabric
Materials
Perhydrolase (S54V variant of M smegmatis, containing 1.5 mg/g active protein)
is used in this
Example. Hydrogen peroxide (analytical grade; 30% w/w) and propylene glycol
diacetate
(>99.7%) are purchased from Sigma Aldrich. Standard dyed fabrics are obtained
from the
Center for Test Materials, Vlaardingen, The Netherlands. The fabrics are dyed
with one of the
dyes (or combinations of dyes) listed in Table 15, which are identified by
their Color Index (CI)
number according to the Society of Dyers and Colourists (UK) and by the
American
Association of Textile Chemists and Colorists (USA).
Procedure
2 each swatches of dyed fabric (see Table 9) of approximately 12.5 cm x 12.5
cm are treated in
a Launder-O-Meter (Rapid Laboratory Dyeing Machine type H 12) according to the
following
procedure:
= 3 each 450-ml stainless steel reaction vessels are filled with 100 ml of
phosphate buffer
(pH 8; 8.9 g/I of disodium phosphate-2H20 + 1.06 g/L of monosodium phosphate
anhydrous).
= 2 each standard fabric swatches are added to each vessel.
= 6.0 ml/L H202 solution (30% wt/wt) and 3.0 ml/L of PGDA (>99.7%) are added
to 2 each
reaction vessels ("perhydrolase" and "blank"). Buffer only is added to the
third reaction
vessel.
= 1.0 ml/L perhydrolase is added to the "perhydrolase" reaction vessel.
= The reactions vessels are closed and placed in the Launder-O-Meter pre-
heated to
60 C.
Incubation is carried out for 30 minutes. Following incubation, the swatches
are rinsed by
overflow, spin dried in an AEG IPX4 centrifuge, and dried in a Novotronic T
494 C household
type dryer.
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The resulting experimental samples are designated "perhydrolase" (perhydrolase
+ H202 +
PGDA), "blank" (H202 + PGDA), and "buffer only."
Table 9. Dyes applied to the different fabric swatches
Cl Sulphur Black 1
Cl Sulphur Black 1
Cl Diazo component 13 (Fast Scarlet R) and Cl
Coupler 2 (Naphthol AS)
Cl Direct Black 22
Cl Direct Black 22
Cl Direct Black 22
Cl Reactive Black 5
Cl Reactive Black 5
Cl Reactive Orange 16
Cl Reactive Blue 71
Cl Reactive Blue 19
Mixture of Cl Reactive Orange 107, Reactive
Red 198, and Reactive Black 5
Mixture of Cl Reactive Orange 107, Reactive
Red 198, and Reactive Blue 220
Evaluation of standard fabric swatches
The effect of perhydrolase treatment on the standard fabric swatches is
evaluated using a
Minolta Chromameter CR 310 in the CIE Lab color space with a D 65 light
source.
Measurements are performed before and after perhydrolase (or control)
treatment. Three
measurements are performed on each swatch (6 measurements total for each
experimental
condition) and the results averaged. Total color difference (TCD) is
calculated using the
formula: TCD = 4 (AL)2 + (Aa)2 + (Ab)2. The A values are the difference
between the fabric
before treatment and the fabric after treatment. The TCD* values are the
difference between
the values obtained for the blank values and for the perhydrolase treatment.
The results are shown, below, in the following Tables, which indicate the dyes
applied to the
fabrics.
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Table 10. Cl Sulphur Black 1
Swatch L/a/b TCD TCD*
Fabric before treatment 24.5/0.5/-3.0 --
Only buffer 24.1/0.4/-3.2 0.5
Blank 25.8/0.4/-3.5 1.4
Perhydrolase 41.4/0.5/-2.8 16.7 15.3
Table 11. Cl Sulphur Black 1 + CR resin
Swatch L/a/b TCD TCD*
Fabric before treatment 24.4/0.8/-0.9 --
Only buffer 23.4/0.7/-1.1 1.1
Blank 24.1/0.7/-1.7 0.8
Perhydrolase 34.7/0.2/-3.0 10.4 10.6
Table 12. Mixture of Cl Diazo component 13 and Cl Coupler 2
Swatch L/a/b TCD TCD*
Fabric before treatment 37.8/58.9/28.0 --
Only buffer 37.1/59.5/28.6 1.1
Blank 38.3/60.4/30.4 2.9
Perhydrolase 39.0/60.8/32.3 4.8 2.0
Table 13. Cl Direct Black 22
Swatch L/a/b TCD TCD*
Fabric before treatment 23.2/-0.4/-1.0 --
Only buffer 22.7/-0.6/-1.5 0.7
Blank 23.2/-0.6/-1.6 0.7
Perhydrolase 26.7/-0.3/-8.6 8.3 7.8
Table 14. Cl Direct Black 22 + cationic finish
Swatch L/a/b TCD TCD*
Fabric before treatment 23.6/-0.1/-1.2 --
Only buffer 22.8/0.1/-1.3 0.9
Blank 22.8/-0.2/-1.4 0.9
Perhydrolase 25.8/0.7/-5.5 4.9 5.1
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Table 15. Cl Direct Black 22 + CR resin
Swatch L/a/b TCD TCD*
Fabric before treatment 23.2/-0.2/-1.0 --
Only buffer 22.7/-0.2/-1.2 0.5
Blank 22.9/-0.4/-1.4 0.7
Perhydrolase 24.1/-0.3/-2.9 2.1 1.9
Table 16. Cl Reactive Black 5 (light)
Swatch L/a/b TCD TCD*
Fabric before treatment 44.1/-0.2/-17.7 --
Only buffer 43.7/-0.1/-17.8 0.4
Blank 44.1/0.1/-17.8 0.3
Perhydrolase 47.1/-0.4/-15.0 4.0 4.1
Table 17. Cl Reactive Black 5 (dark)
Swatch L/a/b TCD TCD*
Fabric before treatment 26.1/1.9/-12.8 --
Only buffer 25.5/2.0/-12.7 0.5
Blank 25.5/2.0/-12.7 0.6
Perhydrolase 27.4/1.2/-12.0 1.6 2.1
Table 18. Cl Reactive Orange 16
Swatch L/a/b TCD TCD*
Fabric before treatment 60.0/50.5/35.8 --
Only buffer 59.5/50.4/35.3 0.7
Blank 59.8/50.6/35.3 0.6
Perhydrolase 61.1/49.5/34.6 1.9 1.8
Table 19. Cl Reactive Blue 71
Swatch L/a/b TCD TCD*
Fabric before treatment 64.1/-21.3/-35.3 --
Only buffer 63.3/-21.3/-36.0 1.0
Blank 63.4/-21.2/-35.7 0.8
Perhydrolase 64.0/-22.3/-33.6 2.0 2.4
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Table 20. Cl Reactive Blue 19
Swatch L/a/b TCD TCD*
Fabric before treatment 41.4/11.3/-42.8 --
Only buffer 40.6/11.7/-43.4 1.1
Blank 40.9/11.8/-43.6 1.1
Perhydrolase 41.4/10.9/-41.9 1.0 1.9
Table 21. Mixture of Cl Reactive Orange 107, Reactive Red 198, and Reactive
Black 5
Swatch L/a/b TCD TCD*
Fabric before treatment 32.3/4.7/3.2 --
Only buffer 31.6/4.6/3.1 0.7
Blank 31.6/4.9/3.4 0.7
Perhydrolase 33.7/5.2/5.3 2.7 2.9
Table 22. Mixture of Cl Reactive Orange 107, Reactive Red 198, and Reactive
Blue 220
Swatch L/a/b TCD TCD*
Fabric before treatment 41.2/14.0/7.6 --
Only buffer 39.7/14.0/8.2 1.6
Blank 40.3/14.0/8.0 1.0
Perhydrolase 41.8/12.9/6.2 1.9 2.6
Perhydrolase-mediated decolorization is observed for all the dyed swatches
tested. These
results demonstrate that enzymatic decolorization is effective for use with a
wide range of
dyes.
Although the foregoing invention has been described in some detail by way of
illustration and
examples for purposes of clarity of understanding, it will be apparent to
those skilled in the art
that certain changes and modifications may be practiced without departing from
the spirit and
scope of the invention. Therefore, the description should not be construed as
limiting the
scope of the invention, which is delineated by the appended claims.
All publications, patents, and patent applications cited herein are hereby
incorporated by
reference in their entireties for all purposes and to the same extent as if
each individual
publication, patent, or patent application were specifically and individually
indicated to be so
incorporated by reference.
