Note: Descriptions are shown in the official language in which they were submitted.
CA 02239335 1998-06-02
WO 97/23606 PCT/US96/20549
Enzyme containing coated granules
Field of the invention
This invention relates to improvements in or relating to enzyme granules, as
well as
s improved processes for producing such granules.
Background of the Invention
Recently the use of enzymes, especially of microbial origin, has been more and
more
common. Enzymes are used in several industries including, for example, the
starch industry,
the dairy industry and the detergent industry. It is well known in the
detergent industry that
the use of enzymes, particularly proteolytic enzymes, has created industrial
hygiene
concerns for detergent factory workers, particularly due to the health risks
associated with
dustiness of the available enzymes.
Since the introduction of enzymes into the detergent business, many
developments in
15 the granulation and coating of enzymes to reduce enzyme dust have been
offered by the
industry. However, in today's state of ever-increasing environmental concern
and
heightened awareness of industrial hygiene, there remains a continuing need
for low dust
enzyme granules. Furthermore, there are additional characteristics desirable
in enzyme
granules not currently available in known granulation products. Some of these
additional
2o characteristics are related to the need to further alleviate industrial
hygiene concerns (lower
dust granules) while optimizing customer and end-user satisfaction
(oxidatively stable and
low residue granules) with the product while simultaneously reducing the cost
of granulation
(improved processing time), thus reducing cost of the overall enzyme product.
Depending on the desired end use of the granule, desirable characteristics may
as include the need for delayed release of the enzyme, preferably without
having to increase
the amount of chlorine scavenger additives currently used in granulation
techniques. This
delayed release has potential benefit, for example, in protecting enzymes from
oxidation or
autolytic degradation in washing machines until sufficient amounts of
stabilizing proteins or
peptides are released from dirty clothing into the wash water. Conversely, if
the granule is to
so be used in an automatic dishwashing detergent (ADD), it would be desirable
to have quick
dissolution of the granule with no residue or film-forming properties on the
surface of the dish
or glassware in the dishwasher. Additional desirable characteristics include
low residue
granule formulations (where low residue is defined as a reduced tendency to
leave
noticeable undissolved residues or filming on clothes or other material such
as glassware or
ss plates, etc.). This characteristic is desirable to the customer (end-user)
of a detergent or
ADD formulation. In addition, improved stability (enhanced shelf life)
formulations are
needed in the industry. Accomplishing all these desired characteristics
simultaneously while
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2
maintaining cost containment for the granule production is a
particularly challenging task. For example, many potential
polymers to delay the release of the enzyme leave behind
insoluble residues, which are undesirable to the user, or
such polymers cause increased processing time, which causes
increased costs. Also, most potential granulating cores,
which are attrition-resistant and therefore suitable for
producing low dust granules, tend to leave behind insoluble
residues.
Therefore, it is an object of the present
invention to provide low dust, low residue enzyme granules.
These granules preferably have increased stability. It is
another object of the present invention to provide processes
and enzyme granule compositions which afford the formation
of such improved granules in much lower processing time,
thus reducing cost of the granular product.
Summary of the Invention
According to one aspect of the present invention,
there are provided improved enzyme-containing granules, such
granules comprising:
a) a core comprising one or more water soluble or
dispersible agent(s), said core being optionally coated with
a vinyl polymer or vinyl copolymer;
b) an enzyme layer comprising one or more
enzymes) and one or more vinyl polymers) or copolymer(s);
and
c) an outer coating comprising one or more
polymers) or copolymers) and, optionally, a low residue
pigment and/or a lubricant.
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In a preferred embodiment of the enzyme granule of
the present invention, the vinyl polymer useful in the core
is a polyvinyl alcohol (PVA), most preferably partially
hydrolyzed PVA, while the vinyl polymer in the enzyme is
polyvinyl pyrrolidone (PVP) and the polymer in the outer
coating is PVA, PVP and/or polyethylene glycol (PEG),
including mixtures thereof.
In a particularly preferred aspect of the
invention, there is provided an enzyme-containing granule
comprising: a) a core comprising a water soluble or
dispersible material coated with a polyvinyl alcohol or
copolymer thereof; b) an enzyme layer comprising one or more
enzymes) and polyvinyl pyrrolidone; and c) an outer coating
layer comprising one or more polymers) selected from the
group consisting of polyvinyl pyrrolidone, polyvinyl alcohol
and polyethylene glycol.
In a further preferred embodiment of the present
invention, the core material is a nonpareil (sugar or salt)
which has been coated with partially hydrolyzed PVA. In
another embodiment of the present invention, the coating on
the core may comprise additional agents such as a
plasticizer.
The enzyme-containing granules of the present
invention may comprise any enzyme; however, in a preferred
embodiment of the present invention, the enzyme is selected
from the group consisting of proteases, amylases, lipases,
cellulases (or components thereof) or oxidases, or mixtures
thereof .
In a preferred embodiment of the present granule,
the enzyme layer comprises a PVP, either alone or in
CA 02239335 2005-02-25
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3a
combination with additional agents such as plasticizers or
anti-agglomeration agents.
For use in ADD applications where quick
dissolution and low filming (low residue) characteristics
are necessary, in addition to low dust, the enzyme-
containing granules of the present invention preferably
comprise an outer coating of PVA, PVP and/or PEG, or
mixtures of such polymers. More preferably, the outer
coating comprises an integral mixture of PVA and PVP, or a
mixture of PVP and PEG, along with a low residue pigment and
a lubricant.
The present invention also comprises an enzyme-
containing granule comprising: a) a nonpareil core; b) an
enzyme layer comprising one or more enzymes) and polyvinyl
pyrrolidone; and c) an outer coating layer comprising a
mixture of polyvinyl alcohol and polyvinyl pyrrolidone or a
mixture of polyvinyl pyrrolidone and polyethylene glycol.
The present invention further comprises an enzyme-
containing granule comprising: a) a nonpareil sugar or salt
core coated with partially hydrolyzed polyvinyl alcohol
having low viscosity; b) an enzyme layer comprising one or
more enzymes) and polyvinyl pyrrolidone; and c) an outer
coating layer comprising a mixture of partially hydrolyzed
polyvinyl alcohol and polyvinyl pyrrolidone, a low residue
pigment and a linear primary alcohol of 9-15 carbon atom
chain length alkane or alkene, or an ethoxylate or
ethoxysulfate derivative thereof.
The present invention still further comprises an
enzyme-containing granule comprising: a) a nonpareil sugar
or salt core coated with partially hydrolyzed polyvinyl
alcohol having low viscosity; b) an enzyme layer comprising
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one or more enzymes) and polyvinyl pyrrolidone; and c) an
outer coating layer comprising a mixture of polyvinyl
pyrrolidone and polyethylene glycol, a low residue pigment
and a linear primary alcohol of 9-15 carbon atom chain
length alkane or alkene, or an ethoxylate or ethoxysulfate
derivative thereof.
This invention also comprises methods for making
low dust granules. In one method aspect of the present
invention there is provided a method comprising:
a) selecting a core material which is a water
soluble or dispersible agent coated with a suitable vinyl
polymer such as PVA;
b) coating the core of step a) with one or more
enzymes) and one or more suitable vinyl polymers) or
copolymer(s); and
c) coating the product of step b) with one or more
suitable polymers) or copolymer(s), alone or in combination
with a low residue pigment or a lubricant, or a mixture
thereof.
In a preferred method embodiment of the present
invention, the vinyl polymer used in step b) of the process
is a PVP and the polymer in step c) is one or more of PVP,
PVA or PEG (or mixtures thereof).
In a particularly preferred method of the present
invention, there is provided a method for making an enzyme-
containing granule, the method comprising: a) selecting a
suitable core material; b) coating the core material of step
a) with an enzyme layer comprising one or more enzymes) and
polyvinyl pyrrolidone; c) coating the product of step b)
with one or more of polyvinyl alcohol, polyvinyl pyrrolidone
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3c
and polyethylene glycol, alone or in combination with one or
more pigment(s), lubricant(s), or a mixture thereof.
In a more preferred embodiment of the present
invention, the method comprises selecting a core coated with
PVA, utilizing PVP in the enzyme layer, and including PVA,
PVP and/or PEG in the outer coating layer. Most preferably,
the outer coating layer further comprises a lubricant such
as an ionic or nonionic surfactant.
Brief Description of the Drawings
Figure 1 is a cross-sectional diagram of an enzyme
granule.
Figure 2 is a cross-sectional diagram of an enzyme
granule comprising additional layers.
Figure 3 is a graph showing dissolution profiles
of certain enzyme granules.
Figure 4 shows dissolution profiles of enzyme
granules comprising various ratios of polymer:pigment and
polymer: pigment: lubricant.
Detailed Description of the Invention
Surprisingly, it has been found that the
incorporation of a vinyl polymer or copolymer, or a mixture
of vinyl polymers and/or other polymers, and preferably PVA,
PVP and PEG, in one or more of the granule layers provides a
granule having improved characteristics such as low dust,
low residue (upon dissolution), altered enzyme release and
increased stability. It
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has also been found that such improved granules can be made in a much reduced
processing time.
A preferred vinyl polymer useful in the present invention is polyvinyl alcohol
{PVA),
which is defined as a homopolymer or copolymer in which vinyl acetate is a
starting monomer
s unit and in which most or all (70-100%) of the acetate moieties are
subsequently hydrolyzed
to alcohol moieties. Other vinyl polymers which may be useful in the present
invention
include, but are not limited to, polyvinyl acetate and polyvinyl pyrrolidone.
Copolymers such
as PVA-methylmethacrylate copolymers or vinyl acetate-vinyl pyrrolidone
copolymers (such
as Luviskol~ VA commercially available from BASF) may also be used in the
present
~o invention. PVA is commercially available in a wide range of molecular
weights, viscosities
and varying degrees of hydrolysis from the polyvinyl acetate precursor. Table
A sets forth
the parameters for categorizing PVA based on these various characteristics.
Table A
~s Grades of PVA Commercially Available
Degree of Viscosity
Viscosity Centipoise Molecular Weight (MW)
ultra low 3-5 5,000-25,000
low 5-15 25, 000-50, 000
2o medium 15-30 50,000-150,000
hig h 30-70 100, 000-200 , 000
Degree of
Hydrolysis % Hydrolysis
25 partially 70-90
intermediately 90-98
fully 98-99
super 99-100
so Any of the PVAs listed in Table A may be used in the present invention.
The type of PVA used will depend in part on which layer of the granule the PVA
is
being used in, and will also depend on what characteristics of the granule are
to be affected.
For example, if PVA is used in the core, it is preferably partially hydrolyzed
PVA and low
viscosity (low molecular weight) because this will result in lower residue
upon dissolution of
35 the granule such as in a washing liquor. The PVA preferred for the enzyme
layer is an
intermediately, fully or super hydrolyzed PVA with low to medium viscosity. In
addition, it is '
contemplated that mixtures of PVA may be used in any or all layers of the
granules of the
present invention.
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As noted, other vinyl polymers are useful in the present invention including,
for
example, PVP of varying molecular weights as described in Table B.
~ Table B
s Grades of PVP Commercially Available
' Commercial Product Molecular Weight
(Luviskol~ K)
9,000 K-17
45,000 K-30
to 350,000 K-60
900,000 K-80
'! ,200, 000 K-90
For purposes of toughness of the granule, high MW PVP is preferred, however,
from
~s a processing and cost perspective, lower MW polymers (9,000-45,000 MW) are
advantageous (while maintaining the performance of the granule).
Additionally, polymers such as polyethylene glycol of MW from about 400-8000,
including PEG 3350, can be used. PEG is commercially available from a number
of
suppliers, including Dow Chemical Company. These polymers, PVP and/or PEG may
be
2o preferable when rapid dissolution is required with little or no film
forming on the surface of the
material coming in contact with the dissolved granule (i.e., dish or glass
surface in
dishwasher).
Cores
The core particles suitable for use in the present invention are preferably of
a highly
2s hydratable material, i.e., a material which is readily dispersible or
soluble in water. The core
material should either disperse (fall apart by failure to maintain its
integrity when hydrated) or
solubilize by going into a true aqueous solution. Clays (bentonite, kaolin),
nonpareils and
agglomerated potato starch are considered dispersible. Nonpareils are
spherical particles
consisting of a seed crystal that has been built onto and rounded into a
spherical shape by
so binding layers of powder and solute to the seed crystal in a rotating
spherical container.
Nonpareils are typically made from a combination of a sugar such as sucrose,
and a powder
such as corn starch. Alternate seed crystal materials include sodium chloride
or sulfate
seeds and other inorganic salts which may be built up with ammonium sulfate,
sodium
sulfate, potassium sulfate and the like.
3s Particles composed of inorganic salts and/or sugars and/or small organic
molecules
may be used as the cores of the present invention. Suitable water soluble
ingredients for
incorporation into cores include: sodium chloride, ammonium sulfate, sodium
sulfate, urea,
citric acid, sucrose, lactose and the like. Water soluble ingredients can be
combined with
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water dispersible ingredients. Cores can be fabricated by a variety of
granulation techniques
including: crystallization, precipitation, pan-coating, fluid-bed coating,
rotary atomization,
extrusion, spheronization and high-shear agglomeration.
The cores of the present invention may further comprise one or more of the
following:
s fillers, plasticizers or fibrous materials. Suitable fillers useful in cores
of the present invention
include inert materials used to add bulk and reduce cost, or used for the
purpose of adjusting
the intended enzyme activity in the finished granulate. Examples of such
fillers include, but
are not limited to, water soluble agents such as urea, salts, sugars and water
dispersible
agents such as clays, talc, silicates, carboxymethyl cellulose or starches.
~o Suitable plasticizers useful in the cores of the present invention are
nonvolatile
solvents added to a polymer to reduce its glass transition temperature,
thereby reducing
brittleness and enhancing deformability. (The glass transition temperature, or
Tg, represents
the onset of segmental mobility for a polymer.) Typically, plasticizers are
low molecular
weight organic compounds and are highly specific to the polymer being
plasticized.
~s Examples include, but are not limited to, polyols (pofyhydric alcohofs, for
example, alcohols
with many hydroxyl radical groups such as glycerol, ethylene glycol, propylene
glycol or
polyethylene glycol), polar low molecular weight organic compounds such as
urea, or other
known plasticizers such as dibutyl or dimethyl phthalate, or water.
Suitable fibrous materials useful in the cores of the present invention
include
ao materials which have high tensile strength and which can be formed into
fine filaments
having a diameter of 1 to 50 microns and a length equal to at least four
diameters. Typical
fibrous materials include, but are not limited to: cellulose, glass fibers,
metal fibers, rubber
fibers, azlon (manufactured from naturally occurring proteins in corn, peanuts
and milk) and
synthetic polymer fibers. Synthetics include Rayon~, Nylon~, acrylic,
polyester, olefin,
2s Saran~, Spandex~ and Vinal~. Typical cellulose fibers would have an average
fiber length
of 160 microns with a diameter of about 30 microns.
In a granule embodiment of the present invention, the core is a water soluble
or
dispersible nonpareil (eitP~er sugar or salt as described above) which can be
further coated
by or built up from the seed crystal (nonpareil) using PVA either alone or in
combination with
sa anti-agglomeration agents such as titanium dioxide, talc, or plasticizers
such as sucrose or
polyols. The PVA may be partially hydrolyzed PVA, intermediately hydrolyzed
PVA, fully
hydrolyzed PVA {all as defined above), or a mixture thereof, with a low to
high degree of
viscosity. Preferably, the nonpareil is coated with partially hydrolyzed PVA,
either alone or in
combination with sucrose or such other plasticizer as known in the art.
Partially hydrolyzed
ss PVA is prefen-ed because it is believed to result in a lower amount of
residue upon
dissolution of the granule than fully hydrolyzed PVA. The level of PVA in the
coating of the
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-- 7 --
nonpareil may represent from about 0.5% to 20% of the weight of the coated
nonpareil. The
core of the granules of the present invention, including any coating on such
core material as
described above, preferably comprises between about 30-85% by weight of the
entire coated
granule.
In a process embodiment of the present invention, the core material, which may
be
any material described herein, is charged into the granulator for coating with
the first layer,
i.e., the enzyme layer.
Enzymes
Any enzyme or combination of enzymes may be used in the present invention.
~o Enzymes are typically coated from relatively impure solutions or slurries
in which the active
enzyme constitutes only a portion of the total dissolved and suspended solids.
Other
suspended solids present in the fermentation broth include other proteins,
peptides,
carbohydrates, other organic molecules and salts. Preferred enzymes include
those
enzymes capable of hydrolyzing substrates, e.g., stains. These enzymes are
known as
~s hydrolases, which include, but are not limited to, proteases (bacterial,
fungal, acid, neutral or,
alkaline), amylases (alpha or beta), lipases, cellulases, and mixtures
thereof. Particularly
preferred enzymes are subtilisins and cellulases. Most preferred are
subtilisins such as
described in US RE 34,606, EP Patent 130 756 B1 and EP Patent Application WO
91/06637, and cellulases or cellulase
2o components isolated from Trichoderma reesei such as Cellulase 123T"" and
MultifectT"" L250,
commercially available from Genencor International, or mixtures thereof, or
those described
in commonly owned PCT Application PCT/US91I07269:
The enzyme layer of the present invention contains, in addition to the enzyme
per se,
a vinyl polymer and preferably PVP. This polymer allows rapid release cf the
enzyme, while
2s avoiding undesirable residue or filming which is common with many polymers.
In a preferred
embodiment of the present invention, the enzyme layer comprises PVP of various
MW PVP
polymers (such as those in Table B above), which PVP is used in an amount of
about 0.1-
5.0% (preferably 1 %) of the granule weight.
The enzyme layer may also further comprise plasticizers and anti-agglomeration
3o agents. Suitable plasticizers useful in the present invention include
polyols such as sugars,
sugar alcohols or polyethylene glycols (PEGs) having a molecular weight less
than 1000,
ureas or other known plasticizers such as dibutyl or dimethyl phthalate, or
water. Suitable
anti-agglomeration agents include fine insoluble material such as talc, TiOz,
clays and
amorphous silica.
The enzyme layer of the present invention, including any nonenzyme solids and
PVP
therein, comprises between about 5%-70% by weight of the coated granule.
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Coating Layers
The granules of the present invention may comprise one or more coating layers.
For
example, such coating Payers may be one or more intermediate coating layers,
or such
coating layers may be one or more outside coating layers, or a combination
thereof. Coating
s layers may serve any of a number of functions in a granule composition,
depending on the
end use of the enzyme granule. For example, coatings may render the enzyme
resistant to
oxidation by bleach, or coating layers may bring about the desirable rate of
dissolution upon
introduction of the granule into an aqueous medium, or provide a barrier
against ambient
moisture in order to enhance the storage stability of the enzyme and reduce
the possibility of
~a microbial growth within the granule.
In an embodiment of the present invention, the outer coating layer comprises
one or
more vinyl polymers) andlor one or more polymers) and, optionally, a low
residue pigment
or other excipients such as lubricants. Such excipients are known to those
skilled in the art.
Furthermore, coating agents may be used in conjunction with other active
agents of the
~s same or different categories.
Suitable vinyl polymers include PVA and/or PVP or mixtures of both. If PVA is
used,
it may be partially hydrolyzed, fully hydrolyzed or intermediately hydrolyzed
PVA having low
to high degrees of viscosity (preferably partially hydrolyzed PVA having low
viscosity). Other
vinyl polymers which may be useful include polyvinyl acetate and polyvinyl
pyrrolidone.
go Useful copolymers include, for example, PVA-methylmethacrylate copolymer.
Other
polymers such as PEG may also be used in the outer layer.
Preferred coatings, particularly for granules needing low dust, rapid
dissolution and
no filming or residue on surfaces, include a mixture of PVP and PVA or a
mixture of PVP and
PEG.
as The coating layers of the present invention may further comprise one or
more of the
following: plasticizers, pigments, lubricants such as surfactants or
antistatic agents and,
optionally, additional enzymes. Suitable plasticizers useful in the coating
layers of the
present invention are plasticizers including, for example, polyols such as
sugars, sugar
alcohols or polyethylene glycols (PEGs) having a molecular weight less than
1000, ureas or
so other known plasticizers such as dibutyl or dimethyl phthalate, or water.
Suitable pigments
useful in the coating layers of the present invention include, but are not
limited to, finely
divided whiteners such as titanium dioxide or calcium carbonate, or colored
pigments, or a
combination thereof. Preferably such pigments are low residue pigments upon
dissolution.
As used herein "lubricants" mean any agent which reduces surface friction,
lubricates
ss the surtace of the granule, decreases static electricity or reduces
friability of the granules.
Lubricants can also play a related role in improving the coating process, by
reducing the
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tackiness of binders in the coating. Thus, lubricants can serve as anti-
agglomeration agents
and wetting agents.
In a preferred embodiment of the present invention, from both a granule and
processing perspective, the outer coating layer comprises a lubricant. The
lubricant reduces
attritional dust even further than the coating alone, dramatically decreases
processing time
and also improves solubility of the granule. It is contemplated that the
lubricant added to the
outer coating may comprise or replace at least about 30% of the polymer or
pigment used in
the coating. In a more preferred embodiment, the lubricant is added to the
granule as an
integral mixture of pigmentlpolymeNlubricant. As used herein, "integral
mixture" means a
~o layer resulting from coating well mixed solutions of the components
(pigment/polymer/lubricant) as opposed to the separate addition (layered
addition) of each
component. As used herein, "pigment" means low residue pigment, such as
titanium dioxide,
and "polymer' means a polymer, vinyl polymer or copolymer, as defined herein,
and
preferably PVP, PVA andlor PEG or a mixture thereof.
~s Suitable lubricating agents include, but are not limited to, surfactants
(ionic, nonionic
or anionic), fatty acids, antistatic agents and antidust agents. Preferably
the lubricant is a
surfactant, and most preferably is an alcohol-based surtactant such as a
linear, primary
alcohol of a 9 to 15 carbon atom chain length alkane or alkene or an
ethoxylate or
ethoxysulfate derivative thereof. Such surfactants are commercially available
as the
2o Neodol~ product line from Shell International Petroleum Company. Other
suitable lubricants
include, but are not limited to, antistatic agents such as StaticGuardTM,
DowneyT"", Triton M
X100 or 120 and the like, antidust agents such as TefIonTM and the like, or
other lubricants
known to those skilled in the art.
The outer coating layer of the present invention preferably comprises between
about
2s 1-20% by weight of the coated granule.
Other Adjunct Ingredients
Adjunct ingredients may be added to the enzyme granules of the present
invention.
Adjunct ingredients may include: metallic salts, solubilizers, activators,
antioxidants, dyes,
inhibitors, binders, fragrances, enzyme protecting agentslscavengers such as
ammonium
so sulfate, ammonium citrate, urea, guanidine hydrochloride, guanidine
carbonate, guanidine
sulfonate, thiourea dioxide, monethyanolamine, diethanolamine,
triethanolamine, amino
acids such as glycine, sodium glutamate and the like, proteins such as bovine
serum
albumin, casein and the like, etc., surfactants, including anionic
surfactants, ampholytic
surfactants, nonionic surfactants, cationic surfactants and long-chain fatty
acid salts,
a~ builders, alkalis or inorganic electrolytes, bleaching agents, bluing
agents and fluorescent
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1 p __
dyes, and caking inhibitors. These surfactants are all described in commonly
assigned PCT
Application PCT/US92/00384.
The granules described herein may be made by methods known to those skilled in
the art of enzyme granulation, including fluidized bed spray-coating, pan-
coating and other
techniques for building up a granule by adding consecutive layers on top of a
starting core
material.
The following examples are representative and not intended to be limiting. One
skilled in the art could choose other enzymes, cores, particles, methods and
coating agents
based on the teachings herein.
~o
Experimental
Example 1
A batch of PVA/sucrose coated nonpareils was produced by coating a PVA/sucrose
solution onto a standard batch of nonpareils. i 00 pounds of -25/+40 mesh
sucrose/starch
~s nonpareils were charged into a 200 Ib capacity coating pan rotating at 45
rpm and heated to
a bed temperature of 150 to 170°F. A coating solution was prepared by
mixing 112 ibs of an
TM
18~o w/w solution of partially hydrolyzed PVA with low viscosity (Airvol 705S,
commercially
available from Air Products, Inc.) with 144 Ibs of a 67% sucrose solution. A
total of 38.4 Ibs
of this unheated mixture were pumped onto the uncoated nonpareils over a
period of twelve
zo hours, providing a coating composed of 2.6% wlw PVA and 12.4°Yo wlw
sucrose, on the basis
of the final product weight. This material was screened to -20/+45 mesh,
yielding 101 Ibs of
usable product and 15 Ibs of scrap. A 20 minute Heubach attrition test on 13.5
mgs of
coated nonpareil cores (prior to enzyme application) resulted in a total dust
reading of 4.2
mg.
2s In a Glatt GPCG-5 fluidized bed spray-coater, 6300 grams of PVA/sucrose
coated
nonpareil cores were charged and fluidized to a bed temperature of
44°C. 11.62 kg of
protease ultrafiltration concentrate produced from B. subfilis, at a
concentration of 5.27% wlw
protease and 25.7% w/w total solids (such that protease represented 20.5% of
total feed
concentrate solids), were mixed with a 1.53 kg solution of a 10% wlw fully
hydrolyzed PVA
3o with low viscosity (Elvanol 90-50, commercially available from E.I. du Pont
de Nemours and
TM
Co., Inc.), and 153 grams of amorphous silica (Zeothix 265, commercially
available from J.M.
Huber Corporation). The enzyme concentrate contained 0.25% sorbitol and 0.5%
sodium
benzoate as formulation chemicals. This enzymeIPVA mixture was then sprayed
onto the
fluidized cores at a starting rate of 40 glminute, romping up to 110 g/minute
over a three hour
35 period, resulting in a weight gain of 3.28 kg. The bed temperature was
gradually reduced
,CA 02239335 2005-02-25
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1 -_
from 46 to 37°C, and the inlet temperature was held at about 57 to
60°C over the course of
the feed ramp. Atomization air pressure was held at 4.0 bar.
After enzyme application, 7.65 kg of a 40% w/w solution of ammonium sulfate
was
sprayed onto the granules, at similar conditions to enzyme application, but at
an atomization
pressure of 3.5 bar. This added another 3.05 kg to the weight of granules. The
mass
balance of the solids weight gain for these two steps was 99.8%. Finally, a
protective
coating solution was applied, made by suspending 765 grams of titanium dioxide
in 1.147 kg
water, then adding 5.10 kg of a 15~o partially hydrolyzed PVA with low
viscosity (Elvanol 51-
05) stock solution, to provide 6.95 kg of a suspension with net 11 % w/w PVA
and 11 % wlw
~o Ti02 concentrations. The coating suspension was sprayed onto the ammonium
sulfate
coated granule at rates of 50-80 g/minute, an inlet temperature of 63 to
67°C, an outlet
temperature of 45 to 49°C, and an atomization air pressure of 4.0 bar.
The final product was
harvested at 13.285 kg, representing a 78% mass balance for the final coating
step, and an
overall 89% mass balance for all spray-coating steps. In terms of percent
weight gain, the
~s enzyme layer representeri a 52% weight gain over the starting core, and the
combined three
layers represented a 119% weight gain over the core. Product was screened
through a 20
mesh screen to remove any agglomerates.
Example 2
Two separate lots of an identical enzyme granule formulation were made in a
Glatt
2o Uniglatt laboratory fluidized bed spray-coater. Processes for the two tots
were virtually
identical, so only the second run is described. The starting material was made
by charging
595 grams of -201+50 mesh PVA coated nonpareils into the fluidized bed. These
cores were
coated by a process similar to that described in Example 1, except that the
coating solution
consisted of an 18% PVA solution (Airvol 705S) without any sucrose added, and
the PVA
2s solution was sprayed onto sucrose/starch nonpareils until the applied PVA
coating
represented 18% of the weight of the final coated nonpareil mass. (The 18% PVA-
coated
nonpareils registered 21.0 mg total dust in a Heubach attrition test prior to
addition of
enzyme.) A 436 gram sample of protease concentrate at a 54 gJkg enzyme
concentration
and 26.1 % total solids concentration was mixed with 94 grams of a 10% PVA
(Elvanol 90-50)
so solution. (Thus, the enzyme represented 20.7% of the total solids in the
feed.) The mixture
was spray-coated onto the fluidized cores at a rate of 7 glminute in the
Uniglatt, with inlet
and outlet temperatures of 55°C and 45°C, respectively, and an
atomization air pressure of
4.0 bar.
Once the enzyme was applied, 588 grams of a 40% ammonium sulfate solution and
~s 539 grams of a suspension containing 11 % PVA (Elvanol 51-05) and 11 % Ti02
were applied
under similar process conditions, with coating rates of 17 glminute and 7
g/minute,
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respectively. The final product weighed 1023 grams, prior to sieving, a 90%
yield on overall
solids gain. This represents a net weight gain over the core weight of .about
18°Yo for the
enzyme layer and about 72% overall. Product was sieved between 20 and 50 mesh
screens
to remove agglomerates and fines.
s Example 1 and Lot 2 of Example 2 were subjected to several tests and
compared
with a comparable commercial product, Savinase 6.0T (available from Novo
Nordisk Industri
AIS). In a Heubach attrition test, using a fill volume of about 17 cc in a 20
minute test time
with an airflow rate of 20 liters per minute desiccated air, the following
comparative dust
levels were obtained:
Total dust ma) Enzyme dust (mA)
Savinase 5.7 24
Example 1 0.6 7
Example 2, Lot 2 0.6 4
~s In a test for potential residue left by enzyme granules after a standard
wash cycle at
60°F, Savinase 6.0T left a fine white residue on the cloth, indicating
the presence of some
insolubles. Examples 1 and 2 left equivalent or lower levels of residue than
Savinase 6.0T.
Dissolution profiles indicating rates of enzyme release under realistic
detergent
conditions are shown in Figure 3 for both lots of Example 2. These profiles
were generated
2o in a dissolution tester with detergent present at 120°F, 10 grains
per gallon hardness and a
fixed medium stir rate. Activities were measured using a synthetic substrate
rate assay.
Even at these high temperatures, it can be seen that the rate of enzyme
release is delayed.
This can be an advantage in that it allows time for scavenging of residue wash
water chlorine
by the ammonium sulfate in the granule and protein materials released from the
clothing.
zs The delay also protects the enzyme against high temperature autolysis until
released
proteins and peptides are available to inhibit autofysis via peptide
inhibition.
Example 3
A granulated cellulase for textile applications was produced in a Uniglatt
spray-water.
842 grams of -30/+50 mesh regular sucrose/starch nonpareils were charged into
the coater
so and fluidized at a bed temperature of 42 to 48°C. A 321 gram
solution of cellulase
ultrafiltration concentrate from T. reesei, containing 6% wlw protein and 21 %
w/w total solids,
was mixed with a 235 gram solution of 10% fully hydrolyzed PVA (Airvol 107).
This mixture
was sprayed onto the fluidized nonpareils at a rate of about 9 g/minute,
resulting in a weight
gain of 88 grams, or about 10.5% w/w. Over the enzyme/PVA layer, 275 grams of
a coating
3s suspension containing 12.7% PVA (Airvol 205) and 12.7% Ti02 was sprayed on,
bringing the
total granule weight to 1000 grams, a total weight gain of 18.8%. Of the final
granule, 6.0%
w/w was cellulase protein.
.:
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In comparison with a competitive product, Denimax Acid T (commercially
available
'from Novo Nordisk lndustri AIS), Example 3 had identical total Heubach dust,
25 mg in both
cases. The polyvinyl alcohol binder and coating provided the granule of
Example 3 with
superior stability of the cellulase activity at high temperature and high
humidity.
s The cellulase granules produced in this example were evaluated for storage
stability
in comparison with a commercial product, Denimax Acid T (Novo Nordisk Industri
AIS). The
amount of residual activity measured after storage for eight days at
37°C, at low and high
relative humidity was as follows:
Relative Humidity (R.H.)
Residual Activity Residual Activity
at 0% R.H, at 60% R.H.
Example 3 117 . 112
Denimax Acid T 102 26
~s Thus, the use of polyvinyl alcohol in the enzyme layer, and especially in
the outer
coating, confers excellent protection against the destabilizing effects of
high temperature in
combination with high humidity.
Example 4
A granulated detergent lipase was produced in the Uniglatt spray-coater. 456
grams
20 of the PVAlsucrose coated nonpareils described in Example 1 were charged
into the reactor.
1.882 liters of a lipase ultrafiltration concentrate containing 10 g/l_ enzyme
and 16.5% w/w
total solids was sprayed cnto the cores without admixed PVA. Inlet and outlet
temperatures
averaged 60 and 45°C, respectively, allowing a coating rate of about 8
glminute at 4 bar
atomization. A 432 gram suspension of 11 % w/w PVA (Elvanol 51-05) and 11 %
wlw Ti02
2s was sprayed onto the lipase coated cores. The lipase application added 307
grams to the
cores, a 67% weight gain. The final product weight was 808 grams, prior to
screening, a net
77% weight gain. A Heubach attrition test yielded a total dust level of 0.8
mg; no
measurement of active lipase dust content was available.
Example 5
3o A granulated detergent protease was produced in a Glatt fluidized bed spray-
water
substantially as described in Examples 1 and 2; however, the outer coating
formulation
applied to the granulation product was an integral mixture of Ti021PVA/Neodol~
in quantities
provided below:
Dry Weight
3s k~c Solution % On Granule
Ti02 47.5 11.0 5
PVA (51-05) 38.0 8.8 4
Neodol~ 9.5 2.2 1
Water 337.0 78.0 0
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432.0 100.0
The TiO~/PVA/Neodol~ coating mixture described above was applied to 684 kg of
uncoated product at a 10% level to yield 760 kg of final product. The coating
was applied at
s a maximum spray rate of about 1.6 kg/min.
The overall coating time for application of this coating mixture Was measured
and
compared to the overall coating time for previous examples (particularly
Example 1) where a
50-50% mixture of PVA/TiOz and 0% Neodol~ were used. Such comparison showed
that
coating time was reduced by about 50% When a surtactant was incorporated in
the coating
~o mixture as compared to coating time for a pigmentlpolymer layer only.
The total dust for the product of Example 5 was 0.6 mg/13.5 g of product, as
measured by the Heubach dust assay referred to in Example 2. Product wash
performance
and solubility were similar to Examples 1 and 2.
Example 6
Following procedures substantially as described in Examples 1-3, three
granulated
celtulase and protease containing products were produced in a Uniglatt spray-
coater.
The three experimental lots comprised the following compositions, coated on
nonpareil core materials:
ao Active Protease Surfactant Pigm/Polym
Exp. # Cellufase (%) Protease (wt %) (~ %~
(%)
A 6.5 0.56 4.49 3.67
B 6.5 0.52 0 3
14
C 6.5 0.59 * .
6.11*
25
* Surtactant suspended in polymer coating; therefore, total polymer and
surfactant
shown in Pigment/Polymer column
1n Experiment A, the non-enzyme coating comprised individual applications of
3G pigment/polymer, TiOz/PVA (Elvanol 51-05), and surfactant (Triton X120).
Experiment B
utilized no surfactant, only a pigment/polymer coating of Ti02/PVA {Elvanol 51-
05). In
Experiment C, the on-enzyme coating comprised surfactant suspended in the
polymer; thus,
the coating in this experiment was Triton X120/Elvanol 51-05.
The three experimental lots (A, B and C) were tested far total dust level in
the
ss Heubach test referred to in Example 2. The following comparative dust
levels were obtained:
EXp # Average dust (mgV13 5 g
A 42.5
B 255.0
C 4.2
ao
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These results show that the distinct surtactant layer of Experiment A seems to
lower
dust; however, a more dramatic effect of lowering dust is evidenced when the
surfactant is
suspended in the PVA forming an integral mixture (see Experiment C).
In addition to lowering dust levels, it was observed that the addition of the
surfactant
s as an integral mixture of the PVA coating layer, enhanced feed rate; thus
reducing the total
processing time, as compared to feed rate for normal pigment/polymer coatings
(without
surtactant).
Granules made as described herein have improved dust characteristics when
compared to other granules known in the art. These improved dust
characteristics are
1o achieved while other desirable characteristics of the granules, such as
solubility, stability,
delayed release and low residue, are maintained or improved. In addition, in
certain
embodiments of the present invention (i.e., Example 5), feed rate may be
enhanced with
resulting reduction in coating time without adversely affecting the desirable
characteristics of
the claimed granules. Thus, cost may be reduced white enhancing product
characteristics.
Example 7
17.8 kg of an alkaline protease solution of 15% solids was spray coated onto
15.5 kg
of PVA coated non-pareils using a Glatt GPGC-5 fluidized bed such that the
fins! product
contained 4% activity by weight. Included with the enzyme solution was low
molecular
2o weight PVP (Luviskol K-17 commercially available from BASF) in an amount
such that the
final product contained 1 % by weight. It was applied with an inlet
temperature between 82-
92°C and an exhaust temperature of 50-60°C. An atomizing air
pressure of 3.5 bar was
utilized throughout the process. After all of the solutions were applied,
water was sprayed
and the product was dried to completion.
1.01 kg of the previous material was coated with 1.2 kg of a 30% solution of
sodium
sulfate using a Vector FL-1 fluidized bed machine. This solution was applied
such that a
20% final product weight was sodium sulfate. It was spray coated with an inlet
temperature
between 70-85°C and an exhaust temperature of 40-50°C. 50 psi
atomizing air pressure
was maintained throughout the process. When the solution was finished, a small
amount of
so water was applied and the product dried to completion.
- Again in the Vector FL-1, 0.9 kg of solution containing 12.5% titanium
dioxide, 7.6%
low molecular weight PVP (Luviskol K-17), 2.5% polyethylene glycol (MW=3350)
and 1.2%
Neodol 23-6.5 was applied. The solution was applied such that the final
product contained
13.3% by weight of the solids. An inlet temperature of 55-62°C and an
exhaust temperature
of 38-41°C were maintained. 50 psi atomizing air pressure was utilized.
After the solution
was applied, a small amount of water was sprayed and the product was dried to
completion.
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To this a 10% Neodol 23-6.5 solution was applied such that the final product
weight
was 0.25% by weight. The solution was applied with an inlet temperature
between 60-65°C
and an exhaust temperature of 40-50°C. After application of the Neodol
solution, a small
amount of water was applied and the product was dried to completion.
s The end product was then sieved through US standard 20 and 50 mesh screens.
99.9% of the product was contained in this fraction. The resulting product was
tested for
dust levels utilizing a Heubach Type III dustmeter. 13.5 grams of product was
tested using a
air flow rate of 200 deca liters per minute for twenty minutes. The level of
active enzyme
dust liberated was measured by dissolving the dust in buffer and detecting
activity using
~o synthetic AAPF substrate. The active dust level obtained was 31.99 ug per
13.5 g sample.
Example 8
55 kg of a protease solution of 15% solids was spray coated onto 25 kg of PVA
TM
coated non-pareil using a Vector FL-60 fluidized bed such that the final
product contained
4% activity by weight. Included with the enzyme solution was low molecular
weight PVP
~s (Luviskol K-17) in an amount such that the final product contained 1 % by
weight. The
enzyme with PVP was applied with an inlet temperature between about 80-
95°C and an
exhaust temperature of between about 65-70°C. An atomizing air pressure
of 75 psi was
utilized throughout the process. When all of the solution was applied, water
was sprayed
and the product was dried to completion.
2o To this, 37 kg of a 30% solution of sodium sulfate was added. This solution
was
applied such that a 20% final product weight was sodium sulfate. It was spray
coated with
an inlet temperature between about 55-60°C and an exhaust temperature
of between about
35-55°C. Again, 75 psi atomizing air pressure was maintained. When the
solution was
finished a small amount of water was applied and the product dried to
completion.
To this, 33 kg of a solution containing 9.4% titanium dioxide, 7.5% low
molecular
weight PVP (Luviskol K-17), 2.5% partially hydrolyzed polyvinyl alcohol and
2.5% Neodol 23-
6.5 was applied. The solution was applied such that the final product
contained 11.6% by
weight of the solids. An inlet temperature of between about 54-56°C and
an exhaust
temperature of between about 38-44°C were maintained. 75 psi atomizing
air pressure was
3G utilized. Once all of the solution was applied, a small amount of water was
sprayed and the
product was dried to completion.
To this a 10% Neodol 23-6.5 solution was applied such that the final product
weight
was 0.5% by weight. The solution was applied with an inlet temperature between
about 55-
60°C and an exhaust temperature of between about 40-45°C. When
the solution was
35 frnished, a small amount of water was applied and the product was dried to
completion.
.;
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The end product was then sieved through US standard 20 and 50 mesh screens.
94% of the product was contained in this fraction. The resulting product was
tested for
toughness utilizing a Heubach Type III dustmeter as in Example 7. The active
dust level
obtained was 0.65 ug per 13.5 g sample.
