Note: Descriptions are shown in the official language in which they were submitted.
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DETERGENT TABLETS COMPRISING A PECTATE LYASE
Field of the Invention
The present invention relates to a laundry or automatic dishwashing
composition
in the tablet form, comprising a pectate lyase.
Backgiround of the invention
Performance of a detergent product is judged by a number of factors, including
the ability to remove soils, and the ability to prevent the redeposition of
the soils,
or the breakdown products of the soils on the articles in the wash. Therefore,
detergent compositions include nowadays a complex combination of active
ingredients which fulfill certain specific needs. In particular, current
detergent
formulations generally include detergent enzymes providing cleaning and fabric
care benefits.
Removal of stains stemming from plants, wood, mould-clay based soils, muddy
soils, and fruits is one of today's toughest cleaning task; especially with
the trend
toward low wash temperatures. These stains typically contain complex mixtures
of fibrous material based mainly on carbohydrates and their derivatives :
fibres
and cell wall components. Plant based soils are additionally accompanied with
amylose, sugars and their derivatives. Food soils are often difficult to
remove
effectively from a soiled substrate. Highly coloured or "dried-on" soils
derived
from fruit and/or vegetable juices are particularly challenging to remove.
Specific
examples of such soils would include orange juice, tomato juice, banana, mango
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or broccoli soils. Indeed, pectin polymers are important constituents of plant
cell
walls. Pectin is a hetero-polysaccharide with a backbone composed of
alternating
homogalacturonan (smooth regions) and rhamnogalacturonan (hairy regions).
The smooth regions are linear polymers of 1,4-linked alpha-D-galacturonic
acid.
The galacturonic acid residues can be methyl-esterified on the carboxyl group
to
a varying degree, usually in a non-random fashion with blocks of
polygalacturonic
acid being completely methyl-esterified. The substrates on which pectin
containing stains are commonly found can be fabrics, dishware or hard
surfaces.
In addition, the complex nature of everyday "body" soils typically found on
pillow
cases, T-shirts, collars and socks, provides a continuous thorough cleaning
challenge for detergents. These soils are difficult to remove completely due
in
part to their interaction with the pectin components in the primary cell walls
of
cotton fibers comprising cotton containing fabrics, and often residues build
up on
such fabric leading to dinginess and yellowing. Moreover, body fluid stains,
such
as blood and menstrual fluids, are often difficult to remove effectively from
a
soiled item, especially when the stains have been aged. Everyday body soils
are
also found on sanitary and kitchen surfaces such as bathtubs, toilet bowls and
dishware.
Therefore, pectin degrading enzymes are known to provide soil/stain removal
benefits when used in washing and cleaning operations, specifically to provide
the removal of a broad range of plant, dirt, and fruit based stains and
enhance
the body soil cleaning profile of the detergent compositions. By pectin
degrading
enzyme it is meant herein any enzyme which acts to break down pectin
substances and pectin related substances. Pectin degrading enzymes can be
classified according to their preferential substrate, highly methyl-esterified
pectin
or low methyl-esterified pectin and polygalacturonic acid (pectate), and their
reaction mechanism, beta-elimination or hydrolysis. Pectin degrading enzymes
can be mainly endo-acting, cutting the polymer at random sites within the
chain
to give a mixture of oligomers, or they may be exo-acting, attacking from one
end
of the polymer and producing monomers or dimers. Several pectinase activities
acting on the smooth regions of pectin are included in the classification of
enzymes provided by the Enzyme Nomenclature (1992) such as pectate lyase
(EC 4.2.2.2), pectin lyase (EC 4.2.2.10), polygalacturonase (EC 3.2.1.15), exo-
polygalacturonase (EC 3.2.1.67), exo-polygalacturonate lyase (EC 4.2.2.9) and
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exo-poly-alpha-galacturonosidase (EC 3.2.1.82). The pectin degrading enzymes
are natural mixtures of the above mentioned enzymatic activities.
Each type of pectin degrading enzyme has a unique profile of substrate
specificity, activity and stability under different hardness, pH, temperature,
surfactant and other detergent ingredient matrix conditions. Pectin degrading
enzymes are specifically directed to degrade pectin substances and in
particular
plant cell walls. In particular, pectate lyase enzymes are directed to the
cleavage
of a-D-(1,4) glycosidic bonds in poly-D-galacturonans by the mechanism of ~i-
elimination. It is recognised in the art that many pectate lyases are metal
ion
dependent, in particular Calcium dependent. Therefore, such enzymes may be
unstable in a detergent matrix and may lose their activity when calcium is
sequestered by builders also present in the detergent matrix. Furthermore, it
also
known that enzymes lose their maximal activity at high pH in the presence of
an
oxidising agent like bleach and are degraded by proteases. In summary, when
certain pectate lyases are formulated in a detergent matrix comprising high
levels
of builder, alkalinity, a bleaching system and protease, their enzymatic
activity
may be significantly decreased unless specific steps are taken to stabilise
them.
This significantly limits the number of available Pectate Lyases that can be
used
in detergent applications.
It has been surprisingly found that the cleaning benefits of pectate lyase
enzymes can be optimised and maximised with a time controlled release
technology. In particular, the time controlled technology is a tablet wherein
the
pectate lyase is separated from the inhibiting / deactivating other detergent
ingredients in a different product phase having a different solubility in the
wash. It
has been surprisingly found that optimal performance efficiency of the pectate
lyase enzyme can be achieved when said enzyme is incorporated into a tablet
and such system delivers significant soil and stain cleaning benefits. It has
further been found that such time controlled release technology allows a
broader
range of Pectate Lyases to be used, including those that show a high degree of
instability in standard detergent matrices.
Detergent compositions in tablet form are known in the art. It is understood
that
detergent compositions in tablet form hold several advantages over detergent
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compositions in particulate form, such as ease of dosing, handling,
transportation
and storage.
Detergent tablets are most commonly prepared by pre-mixing components of a
detergent composition and forming the pre-mixed detergent components into a
tablet using any suitable equipment, preferably a tablet press. Tablets are
typically formed by compression of the components of the detergent composition
so that the tablets produced are sufficiently robust to be able to withstand
handling and transportation without sustaining damage. In addition to being
robust, tablets must also dissolve sufficiently fast so that the detergent
components are released into the wash water as soon as possible at the
beginning of the wash cycle. Prior art already tackled the problem of finding
a
balance between tablet robustness and tablet dissolution.
One solution has been to design multi-phase tablets. Multi-phase detergent
tablets described in the prior art are prepared by compressing a first
composition
in a tablet press to form a substantially planar first layer. A further
detergent
composition is then delivered to the tablet press on top of the first layer.
This
second composition is then compressed to form another substantially planar
second layer. Other multi-phase tablets exhibiting differential dissolution
are
prepared such that the second layer is compressed at a lower force than the
first
layer resulting in faster dissolution of the second layer.
The use of pectin degrading enzymes in detergents has already been recognised
in the art. The use of pectin degrading enzyme is also recognised for the
cleaning of contact lenses (US 4,710,313 - J60196724). Enzymes having a
pectinase activity are described in DE 36 35 427 to increase the capacity of
the
detergent for removing inorganic dirt, e.g. sludges, from laundry without
damaging the fibres and without discoloration to allow the use of zeolites and
polycarbonate builders which have a lower capacity for dispersing inorganic
materials than the phosphates. Benefits for the use of pectin degrading
enzymes
in detergent formulations, particularly those designed for use in laundry,
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dishwashing and household cleaning operations have been recognised in
W095/25790. JP 60226599 describes detergent compositions comprising
conventional detergent actives and a cellulase and hydroiase such as
hemicellulase, pectinase, amylase or protease. The combination of cellulase
and
hydrolase is said to give a good washing effect on inorganic fouling together
with
enzymatic activity. W095/09909 describes an enzyme preparation comprising
modified enzymes selected from the group of amylase, lipase, oxidoreductase,
pectinase or hemicellulase; the modified enzymes having an improved
performance due to an alkaline pl and/or increased surface activity obtained
by
chemical modification or amino acid substitution. Modified pectin and/or
pectolytic and/or hemi-cellulolytic and /or lipolytic enzymes are applied
advantageously in the papermaking industry and modified amylase and/or lipase
in laundry and dishwashing.
In particular, Pectate lyases have been cloned from different bacterial genera
such as Erwinia, Pseudomonas, Klebsiella and Xanthomonas, Streptomyces,
Penicillium, Baceriodes, Thermomonospora, Fusarium, and Aspergillus. Also
from Bacillus subtilis (Nasser et al. (1993) FEBS 335:319-326) and Bacillus
sp.
YA-14 (Kim et al. (1994) Biosci. Biotech. Biochem. 58:947-949) cloning of a
pectate lyase has been described. Purification of pectate lyases with maximum
activity in the pH range of 8-10 produced by Bacillus pumilus (Dave and Vaughn
(1971) J. Bacteriol. 108:166-174), B. polymyxa (Nagel and Vaughn (1961) Arch.
Biochem. Biophys. 93:344-352), B. stearothermophilus (Karbassi and Vaughn
(1980) Can. J. Microbiol. 26:377-384), Bacillus sp. (Hasegawa and Nagel (1966)
J. Food Sci. 31:838-845) and Bacillus sp. RK9 (Kelly and Fogarty (1978) Can.
J.
Microbiol. 24:1164-1172) has been reported. WO 98/45393 discloses detergent
compositions containing protopectinase with remarkable detergency against
muddy soiling:
However, the formulation of a pectate lyase into a detergent tablet with time
controlled release, for superior cleaning performance, has never been
previously
recognised.
Summaryr of the invention
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The present invention relates to laundry or automatic dishwashing compositions
in the tablet form, comprising a pectate lyase for improved cleaning
performance,
especially on plant-based and body soils.
Detailed description of the invention
The detergent tablet of the present invention is not only sufficiently robust
to
withstand handling and transportation, but also at least a portion of which
dissolves rapidly in the wash water providing rapid delivery of the pectate
lyase
enzyme.
It is preferred that at least one phase of the tablet dissolves in the wash
water
within the first ten minutes, preferably five minutes, more preferably four
minutes
of the wash cycle of an automatic dishwashing or laundry washing machine.
Preferably the washing machine is either an automatic dishwashing or laundry
washing machine. The time within which the mufti-phase tablet or a phase
thereof or a detergent active component dissolves is determined according to
DIN 44990 using a dishwashing machine available from Bosch on the normal
65°C washing program with water hardness at 18°H using a minimum
of six
replicates or a sufficient number to ensure reproducibility.
Preferably, the pectate lyase and buffer materials are incorporated into the
rapid
dissolving portion of the tablet. Without wishing to be bound by theory, it is
believed that the pectate lyase is released earlier than the inhibiting /
deactivating other detergent ingredients and that optimum pectate lyase
activity
is obtained at the beginning of the wash under buffered conditions, allowing
the
formulation in detergent of pectate lyases in the full range of available
pectate
lyases. Also contemplated are tablets wherein the pectate lyase is released at
different stages of the wash process according to the needs of the pectate
lyase
application and matrix conditions.
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Tablet detergent
The present invention encompasses the following different tablet embodiments
(a) a tablet which comprises a section 1 and a section 2 wherein the section 2
comprises a higher level of pectate lyase.
(b) a tablet as described in (a) wherein the tensile strength of section 1 is
larger,
preferably at least 2% larger, more preferably 5%, even more preferably 10%
and most preferably 30%, than the tensile strength of section 2.
(c) a tablet according to (a) or (b) wherein section 2 has a larger exposed
surface
than section 1.
(d) a tablet according to (a), (b) or (c) wherein section 2 has an exposed
surface
equal to the exposed surface of the tablet.
(e) a tablet wherein section 2 is applied by a coating process.
(f) a tablet according to (a) to (e) wherein section 1 is a slow dissolving
section
and section 2 is a rapid dissolving section.
By "slow dissolving" it is meant herein a tablet dissolving in more than 10
minutes
according to the DIN 44990 method described. By "rapid dissolving" it is meant
herein a tablet dissolving within the first ten minutes, preferably five
minutes,
more preferably four minutes according to the DIN 44990 method described
above.
Suitable for the purpose of the present invention are the single and multi-
phase
detergent tablets for use in automatic dishwashing and laundry, having
improved
strength, especially on long term storage and excellent dissolution
characteristics
as described in the co-pending European Application No. 9818716.4 filed 28
August 1998.
Such detergent tablet is not only sufficiently robust to withstand handling
and
transportation, but also at least a portion of which dissolves rapidly in the
wash
water providing rapid delivery of the pectate lyase enzyme and buffer
materials.
It is preferred that at least one phase, preferably section 2, of the tablet
dissolves
in the wash water within the first ten minutes, preferably five minutes, more
preferably four minutes of the wash cycle of an automatic dishwashing or
laundry
washing machine according to DIN 44990, above.
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The detergent tablets of the present invention comprise a first phase and, in
multi-phase tablet embodiments, also comprise a second and optional
subsequent phases. The first phase is in the form of a shaped body of
detergent composition comprising one or more detergent components as
described below. Preferred detergent components of the first phase include
other builder components, bleach, enzymes and surfactant. The components of
the detergent composition are mixed together by, for example admixing dry
components or spraying-on liquid components. The components are then
formed into a first phase using any suitable compression equipment, but
preferably in a tablet press.
In mould embodiments, the first phase is prepared such that it comprises at
least
one mould in the surface of the shaped body. In a preferred embodiment the
mould is created using a specially designed tablet press wherein the surface
of
the punch that contacts the detergent composition is shaped such that when it
contacts and presses the detergent composition it presses a mould, or multiple
moulds into the first phase of the multi-phase detergent tablet. Preferably,
the
mould will have an inwardly concave or generally concave surface to provide
improved adhesion to the second phase.
The tablets of the invention can also include one or more additional phases
prepared from a composition or compositions which comprise one or more
detergent components as described below. At least one phase (herein referred
to as a second phase) preferably takes the form of a particulate solid (which
term
encompasses powders, granules, agglomerates, and other particulate solids
including mixtures thereof with liquid binders, meltable solids, spray-ons,
etc)
compressed either as a layer or into/within the one or more moulds of the
first
phase of the detergent tablet such that the second phase itself takes the form
of
a shaped body. Preferred detergent components include builders, colourants,
binders, surfactants, disrupting agents and enzymes, in particular Pectate
Lyase
enzymes. In another preferred aspect of the present invention the second and
optional subsequent phases comprise a disrupting agent that may be selected
from either a disintegrating agent or an effervescent agent. Suitable
disintegrating agents include agents that swell on contact with water or
facilitate
water influx and/or efflux by forming channels in the detergent tablet. Any
known
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disintegrating or effervescing agent suitable for use in laundry or
dishwashing
applications is envisaged for use herein. Suitable disintegrating agent
include
starch, starch derivatives such as Arbocel (tradename), Vivapur (tradename)
both available from Rettenmaier, Nymcel (tradename) available from Metsa-serla
alginates, acetate trihydrate, burkeite, monohydrated carbonate formula
Na2C03.H20, carboxymethylcellulose (CMC), CMC-based polymers, sodium
acetate, aluminium oxide. Suitable effervescing agents are those that produce
a
gas on contact with water. Suitable effervescing agents may be oxygen,
nitrogen
dioxide or carbon dioxide evolving species. Examples of preferred effervescent
agents may be selected from the group consisting of perborate, percarbonate,
carbonate, bicarbonate in combination with inorganic acids such as sulphamic
acid and/or carboxylic acids such as citric, malic and malefic acid and
mixtures
thereof.
The components of the detergent composition are mixed together by for example
premixing dry components and admixing, preferably by spray-on, liquid
components. The components of the second and optional subsequent phases
are then compressed to form one or more layers or are fed into and retained
within the mould provided by the first phase.
The preferred mould embodiments of the present invention comprises two
phases; a first and a second phase. The first phase will normally comprise one
mould and the second phase will normally consist of a single detergent active
composition. However, it is envisaged that the first phase may comprise more
than one mould and the second phase may be prepared from more than one
detergent active composition. Furthermore, it is also envisaged that the
second
phase may comprise more than one detergent active composition contained
within one mould. It is also envisaged that several detergent active
compositions
are contained in separate moulds. In this way potentially chemically sensitive
detergent components can be separated in order to avoid any loss in
performance caused by components reacting together and potentially becoming
inactive or exhausted.
In a preferred aspect of the present invention the first, second and/or
optional
subsequent phases may comprise a binder. Where present the binder is
selected from the group consisting of organic polymers, for example
polyethylene
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and/or polypropylene glycols having an average molecular weight of from about
1000 to about 12000, especially those of molecular weight 4000, 6000 and 9000,
polyvinyl pyrolindone (PVP), especially PVP of molecular weight 90 000,
polyacrylates, sugars and sugar derivatives, starch and starch derivatives,
for
example hydroxy propyl methyl cellulose (HPMC) and carboxy methyl cellulose
(CMC); and inorganic polymers, such as hexametaphosphate. The
polyethyleneglycol binders are highly preferred herein.
In a preferred aspect of the present invention the first phase constitutes at
least
50% of the total tablet weight. More preferably the first phase comprises from
60
to 90%, even more preferably from 70 to 85% and most preferably from 80 to
85% of the total tablet weight. The second and optional subsequent phases
comprise less than 40% of the tablet weight. More preferably the second and/or
optional subsequent phases comprise between 20 and 30%, most preferably
from 8 to 15% of the total tablet weight
The detergent tablets are prepared using any suitable tabletting equipment.
Preferably multi-phase tablets herein are prepared by compression in a tablet
press capable of preparing a tablet comprising a mould. In a particularly
preferred embodiment of the present invention the first phase is prepared
using a
specially designed tablet press. The punches) of this tablet press are
modified
so that the surface of the punch that contacts the detergent composition has a
convex surface.
A first detergent composition is delivered into the die of the tablet press
and the
punch is lowered to contact and then compress the detergent composition to
form a first phase. The first detergent composition is compressed using an
applied pressure of at least 250 kg/cm2, preferably between 350 and 2000
kg/cm2, more preferably 500 to 1500 kg/cm2, most preferably 600 to 1200
kg/cm2.
The punch is then elevated, exposing the first phase containing a mould. A
second and optionally a subsequent detergent compositions) comprising the
pectate lyase is then delivered into the mould. The specially designed tablet
press punch is then lowered a second time to lightly compress the second and
optional subsequent detergent compositions) to form the second and optional
subsequent phase(s). In another embodiment of the present invention where an
optional subsequent phase is present the optional subsequent phase is prepared
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in a subsequent compression step substantially similar to the second
compression step described above. The second and optional subsequent
detergent compositions) is compressed at a pressure of preferably less than
350
kg/cm2, more preferably from 40 to 300 kg/cm2, most preferably from 70 to 270
kg/cm2. After compression of the second detergent composition, the punch is
elevated a second time and the mufti-phase detergent tablet is ejected from
the
tablet press. Single and mufti-layer tablets without moulds can be prepared in
a
similar manner except using a tablet punch having a planar surface.
The detergent tablets of the invention are prepared by compression of one or
more compositions comprising detergent active components. Suitably, the
compositions may include a variety of different detergent components including
builder compounds, surfactants, enzymes, bleaching agents, alkalinity sources,
colourants, perfume, lime soap dispersants, organic polymeric compounds
including polymeric dye transfer inhibiting agents, crystal growth inhibitors,
heavy
metal ion sequestrants, metal ion salts, enzyme stabilisers, corrosion
inhibitors,
suds suppressers, solvents, fabric softening agents, optical brighteners and
hydrotropes. In the following, the proportions of these active components are
given by weight of the corresponding composition of active detergent
components, unless specified otherwise.
In mufti-phase tablets, highly preferred detergent components of the first
rapid
dissolving phase include builder, enzymes specifically the pectate lyase,
buffering agent and disrupting agent. Highly preferred detergent components of
the second slower dissolving phase include a builder compound, a surfactant,
an
enzyme and a bleaching agent.
Builders
Detergent builders can optionally be included in the compositions herein to
assist
in controlling mineral hardness. Inorganic as well as organic builders can be
used. Builders are typically used in fabric laundering compositions to assist
in
the removal of particulate soils.
The level of builder can vary widely depending upon the end use of the
composition.
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Inorganic or P-containing detergent builders include, but are not limited to,
the
alkali metal, ammonium and alkanolammonium salts of polyphosphates
(exemplified by the tripolyphosphates, pyrophosphates, and glassy polymeric
meta-phosphates), phosphonates, phytic acid, silicates, carbonates (including
bicarbonates and sesquicarbonates), sulphates, and aluminosilicates. However,
non-phosphate builders are required in some locales. Importantly, the
compositions herein function surprisingly well even in the presence of the so-
called "weak" builders (as compared with phosphates) such as citrate, or in
the
so-called "underbuilt" situation that may occur with zeolite or layered
silicate
builders.
Examples of silicate builders are the alkali metal silicates, particularly
those
having a Si02:Na20 ratio in the range 1.6:1 to 3.2:1 and layered silicates,
such
as the layered sodium silicates described in U.S. Patent 4,664,839, issued May
12, 1987 to H. P. Rieck. NaSKS-6 is the trademark for a crystalline layered
silicate marketed by Hoechst (commonly abbreviated herein as "SKS-6"). Unlike
zeolite builders, the Na SKS-6 silicate builder does not contain aluminum.
NaSKS-6 has the delta-Na2Si05 morphology form of layered silicate. It can be
prepared by methods such as those described in German DE-A-3,417,649 and
DE-A-3,742,043. SKS-6 is a highly preferred layered silicate for use herein,
but
other such layered silicates, such as those having the general formula
NaMSixO2x+1 ~yH20 wherein M is sodium or hydrogen, x is a number from 1.9 to
4, preferably 2, and y is a number from 0 to 20, preferably 0 can be used
herein.
Various other layered silicates from Hoechst include NaSKS-5, NaSKS-7 and
NaSKS-11, as the alpha, beta and gamma forms. As noted above, the delta-
Na2Si05 (NaSKS-6 form) is most preferred for use herein. Other silicates may
also be useful such as for example magnesium silicate, which can serve as a
crispening agent in granular formulations, as a stabilizing agent for oxygen
bleaches, and as a component of suds control systems.
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Examples of carbonate builders are the alkaline earth and alkali metal
carbonates as disclosed in German Patent Application No. 2,321,001 published
on November 15, 1973.
Aluminosilicate builders are useful in the present invention. Aluminosilicate
builders are of great importance in most currently marketed heavy duty
granular
detergent compositions, and can also be a significant builder ingredient in
liquid
detergent formulations. Aluminosilicate builders include those having the
empirical formula:
Mz(zA102)y]~xH20
wherein z and y are integers of at least 6, the molar ratio of z to y is in
the range
from 1.0 to about 0.5, and x is an integer from about 15 to about 264.
Useful aluminosilicate ion exchange materials are commercially available.
These
aluminosilicates can be crystalline or amorphous in structure and can be
naturally-occurring aluminosilicates or synthetically derived. A method for
producing aluminosilicate ion exchange materials is disclosed in U.S. Patent
3,985,669, Krummel, et al, issued October 12, 1976. Preferred synthetic
crystalline aluminosilicate ion exchange materials useful herein are available
under the designations Zeolite A, Zeolite P (B), Zeolite MAP and Zeolite X. In
an
especially preferred embodiment, the crystalline aluminosilicate ion exchange
material has the formula:
Nal2I(A102)12(Si02)121~xH20
wherein x is from about 20 to about 30, especially about 27. This material is
known as Zeolite A. Dehydrated zeolites (x = 0 - 10) may also be used herein.
Preferably, the aluminosilicate has a particle size of about 0.1-10 microns in
diameter.
Organic detergent builders suitable for the purposes of the present invention
include, but are not restricted to, a wide variety of polycarboxyiate
compounds.
As used herein, "polycarboxylate" refers to compounds having a plurality of
carboxylate groups, preferably at least 3 carboxylates. Polycarboxylate
builder
can generally be added to the composition in acid form, but can also be added
in
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the form of a neutralized salt. When utilized in salt form, alkali metals,
such as
sodium, potassium, and lithium, or alkanolammonium salts are preferred.
Included among the polycarboxylate builders are a variety of categories of
usefui
materials. One important category of polycarboxylate builders encompasses the
ether polycarboxylates, including oxydisuccinate, as disclosed in Berg, U.S.
Patent 3,128,287, issued April 7, 1964, and Lamberti et al, U.S. Patent
3,635,830, issued January 18, 1972. See also "TMSlTDS" builders of U.S.
Patent 4,663,071, issued to Bush et al, on May 5, 1987. Suitable ether
polycarboxylates also include cyclic compounds, particularly alicyclic
compounds,
such as those described in U.S. Patents 3,923,679; 3,835,163; 4,158,635;
4,120,874 and 4,102,903.
Other useful detergency builders include the ether hydroxypolycarboxylates,
copolymers of malefic anhydride with ethylene or vinyl methyl ether, 1, 3, 5-
trihydroxy benzene-2, 4, 6-trisulphonic acid, and carboxymethyloxysuccinic
acid,
the various alkali metal, ammonium and substituted ammonium salts of
polyacetic acids such as ethylenediamine tetraacetic acid and nitrilotriacetic
acid,
as well as polycarboxylates such as mellitic acid, succinic acid,
oxydisuccinic
acid, polymaleic acid, benzene 1,3,5-tricarboxylic acid,
carboxymethyloxysuccinic
acid, and soluble salts thereof.
Citrate builders, e.g., citric acid and soluble salts thereof (particularly
sodium
salt), are polycarboxylate builders of particular importance for heavy duty
liquid
detergent formulations due to their availability from renewable resources and
their biodegradability. Citrates can also be used in granular compositions,
especially in combination with zeolite and/or layered silicate builders.
Oxydisuccinates are also especially useful in such compositions and
combinations.
Also suitable in the detergent compositions of the present invention are the
3,3-
dicarboxy-4-oxa-1,6-hexanedioates and the related compounds disclosed in U.S.
Patent 4,566,984, Bush, issued January 28, 1986. Useful succinic acid builders
include the C5-C2p alkyl and alkenyl succinic acids and salts thereof. A
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particularly preferred compound of this type is dodecenylsuccinic acid.
Specific
examples of succinate builders include: laurylsuccinate, myristylsuccinate,
palmitylsuccinate, 2-dodecenylsuccinate (preferred), 2-pentadecenylsuccinate,
and the like. Laurylsuccinates are the preferred builders of this group, and
are
described in European Patent Application 86200690.5/0,200,263, published
November 5, 1986.
Other suitable polycarboxylates are disclosed in U.S. Patent 4,144,226,
Crutchfield et al, issued March 13, 1979 and in U.S. Patent 3,308,067, Diehl,
issued March 7, 1967. See also Diehl U.S. Patent 3,723,322.
Fatty acids, e.g., C12-C1g monocarboxylic acids, can also be incorporated into
the compositions alone, or in combination with the aforesaid builders,
especially
citrate and/or the succinate builders, to provide additional builder activity.
Such
use of fatty acids will generally result in a diminution of sudsing, which
should be
taken into account by the formulator.
In situations where phosphorus-based builders can be used, and especially in
the formulation of bars used for hand-laundering operations, the various
alkali
metal phosphates such as the well-known sodium tripolyphosphates, sodium
pyrophosphate and sodium orthophosphate can be used. Phosphonate builders
such as ethane-1-hydroxy-1,1-diphosphonate and other known phosphonates
(see, for example, U.S. Patents 3,159,581; 3,213,030; 3,422,021; 3,400,148 and
3,422,137) can also be used.
Surfactants suitable herein include anionic surfactants such as alkyl
sulfates,
alkyl ether sulfates, alkyl benzene sulfonates, alkyl glyceryl sulfonates,
alkyl and
alkenyl sulphonates, alkyl ethoxy carboxylates, N-acyl sarcosinates, N-acyl
taurates and alkyl succinates and sulfosuccinates, wherein the alkyl, alkenyl
or
acyl moiety is C5-C20 , preferably C10-C1g linear or branched; cationic
surfactants such as choline esters (US-A-4228042, US-A-4239660 and US-A-
4260529) and mono Cg-C1g N-alkyl or alkenyl ammonium surfactants wherein
the remaining N positions are substituted by methyl, hydroxyethyl or
hydroxypropyl groups; low and high cloud point nonionic surfactants and
mixtures thereof including nonionic alkoxylated surfactants (especially ethoxy-
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16
lates derived from Cg-C1g primary alcohols), ethoxylated-propoxylated alcohols
(e.g., Olin Corporation's Poly-Tergent0 SLF18), epoxy-capped
poly(oxyalkylated)
alcohols (e.g., Olin Corporation's Poly-TergentC~ SLF18B - see WO-A-94/22800),
ether-capped poly(oxyalkylated) alcohol surfactants, and block polyoxyethylene-
polyoxypropylene polymeric compounds such as PLURONIC~, REVERSED
PLURONIC~, and TETRONIC~ by the BASF-Wyandotte Corp., Wyandotte,
Michigan; amphoteric surfactants such as the amine oxides and alkyl
amphocarboxylicc surfactants such as MiranolT"" C2M; and zwitterionic
surfactants such as the betaines and sultaines; and mixtures thereof.
Surfactants suitable herein are disclosed, for example, in US-A-3,929,678 , US-
A- 4,259,217, EP-A-0414 549, WO-A-93/08876 and WO-A-93/08874.
Surfactants are typically present at a level of from about 0.2% to about 30%
by
weight, more preferably from about 0.5% to about 10% by weight, most
preferably from about 1 % to about 5% by weight of composition.
Enzymes suitable for use in section 1 herein include enzymes like protease,
amylase, lipase, cutinase and/or cellulase.
Suitable proteases are the subtilisins which are obtained from particular
strains of
B. subtilis and B. licheniformis (subtilisin BPN and BPN'). One suitable
protease
is obtained from a strain of Bacillus, having maximum activity throughout the
pH
range of 8-12, developed and sold as ESPERASE~ by Novo Industries A/S of
Denmark, hereinafter "Novo". The preparation of this enzyme and analogous
enzymes is described in GB 1,243,784 to Novo. Other suitable proteases
include ALCALASE~, DURAZYM~ and SAVINASE~ from Novo and
MAXATASE~, MAXACAL~, PROPERASE~ and MAXAPEM~ (protein
engineered Maxacal) from Gist-Brocades. Proteolytic enzymes also encompass
modified bacterial serine proteases, such as those described in European
Patent
Application Serial Number 87 303761.8, filed April 28, 1987 (particularly
pages
17, 24 and 98), and which is called herein "Protease B", and in European
Patent
Application 199,404, Venegas, published October 29, 1986, which refers to a
modified bacterial serine protealytic enzyme which is called "Protease A"
herein.
Suitable is the protease called herein "Protease C", which is a variant of an
alkaline serine protease from Bacillus in which lysine replaced arginine at
position 27, tyrosine replaced valine at position 104, serine replaced
asparagine
at position 123, and alanine replaced threonine at position 274. Protease C is
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17
described in EP 90915958:4, corresponding to WO 91/06637, Published May 16,
1991. Genetically modified variants, particularly of Protease C, are also
included
herein.
A preferred protease referred to as "Protease D" is a carbonyl hydrolase
variant
having an amino acid sequence not found in nature, which is derived from a
precursor carbonyl hydrolase by substituting a different amino acid for a
plurality
of amino acid residues at a position in said carbonyl hydrolase equivalent to
position +76, preferably also in combination with one or more amino acid
residue
positions equivalent to those selected from the group consisting of +99, +101,
+103, +104, +107, +123, +27, +105, +109, +'126, +128, +135, +156, +166, +195,
+197, +204, +206, +210, +216, +217, +218, +222, +260, +265, and/or +274
according to the numbering of Bacillus amyloliquefaciens subtilisin, as
described
in W095/10591 and in the patent application of C. Ghosh, et al, "Bleaching
Compositions Comprising Protease Enzymes" having US Serial No. 08/322,677,
filed October 13, 1994. Also suitable is a carbonyl hydrolase variant of the
protease described in W095/10591, having an amino acid sequence derived by
replacement of a plurality of amino acid residues replaced in the precursor
enzyme corresponding to position +210 in combination with one or more of the
following residues : +33, +62, +67, +76, +100, +101, +103, +104, +107, +128,
+129, +130, +132, +135, +156, +158, +164, +166, +167, +170, +209, +215,
+217, +218, and +222, where the numbered position corresponds to naturally-
occurring subtilisin from Bacillus amyloliquefaciens or to equivalent amino
acid
residues in other carbonyl hydrolases or subtilisins, such as Bacillus lentus
subtilisin (co-pending patent application US Serial No. 60/048,550, filed June
04,
1997).
Also suitable for the present invention are proteases described in patent
applications EP 251 446 and WO 91/06637, protease BLAP~ described in
W091/02792 and their variants described in WO 95/23221.
See also a high pH protease from Bacillus sp. NCIMB 40338 described in WO
93/18140 A to Novo. Enzymatic detergents comprising protease, one or more
other enzymes, and a reversible protease inhibitor are described in WO
92/03529 A to Novo. When desired, a protease having decreased adsorption
and increased hydrolysis is available as described in WO 95/07791 to Procter &
Gamble. A recombinant trypsin-like protease for detergents suitable herein is
described in WO 94/25583 to Novo. Other suitable proteases are described in
EP 516 200 by Unilever.
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18
The proteolytic enzymes are incorporated in the detergent compositions of the
present invention a level of from 0.0001 % to 2%, preferably from 0.001 % to
0.2%, more preferably from 0.005% to 0.1 % pure enzyme by weight of the
composition.
The cellulases usable in the present invention include both bacterial or
fungal
cellulases. Preferably, they will have a pH optimum of between 5 and 12 and a
specific activity above 50 CEVU/mg (Cellulose Viscosity Unit). Suitable
cellulases
are disclosed in U.S. Patent 4,435,307, Barbesgoard et al, J61078384 and
W096/02653 which discloses fungal cellulase produced respectively from
Humicola insolens, Trichoderma, Thielavia and Sporotrichum. EP 739 982
describes cellulases isolated from novel Bacillus species. Suitable cellulases
are
also disclosed in GB-A-2.075.028; GB-A-2.095.275; DE-OS-2.247.832 and
W095/26398.
Examples of such cellulases are cellulases produced by a strain of Humicola
insolens (Humicola grisea var. thermoidea), particularly the Humicola strain
DSM
1800.
Other suitable cellulases are cellulases originated from Humicola insolens
having
a molecular weight of about 50KDa, an isoelectric point of 5.5 and containing
415
amino acids; and a "43kD endoglucanase derived from Humicola insolens, DSM
1800, exhibiting cellulase activity; a preferred endoglucanase component has
the
amino acid sequence disclosed in PCT Patent Application No. WO 91/17243.
Also suitable cellulases are the EGIII cellulases from Trichoderma
longibrachiatum described in W094/21801, Genencor, published September 29,
1994. Especially suitable cellulases are the cellulases having color care
benefits.
Examples of such cellulases are cellulases described in European patent
application No. 91202879.2, filed November 6, 1991 (Novo). Carezyme and
Celluzyme (Novo Nordisk A/S) are especially useful. See also W091/17244 and
W091/21801. Other suitable cellulases for fabric care and/or cleaning
properties
are described in W096/34092, W096/17994 and W095/24471.
Said cellulases are normally incorporated in the detergent composition at
levels
from 0.0001 % to 2% of pure enzyme by weight of the detergent composition.
Peroxidase enzymes are used in combination with oxygen sources, e.g.
percarbonate, perborate, persulfate, hydrogen peroxide, etc and with a
phenolic
substrate as bleach enhancing molecule. They are used for "solution
bleaching",
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19
i.e. to prevent transfer of dyes or pigments removed from substrates during
wash
operations to other substrates in the wash solution. Peroxidase enzymes are
known in the art, and include, for example, horseradish peroxidase, ligninase
and
haloperoxidase such as chloro- and bromo-peroxidase. Peroxidase-containing
detergent compositions are disclosed, for example, in PCT International
Application WO 89/099813, W089/09813 and in European Patent application EP
No. 91202882.6, filed on November 6, 1991 and EP No. 96870013.8, filed
February 20, 1996. Also suitable is the laccase enzyme.
Enhancers are generally comprised at a level of from 0.1 % to 5% by weight of
total composition. Preferred enhancers are substitued phenthiazine and
phenoxasine 10-Phenothiazinepropionicacid (PPT), 10-ethylphenothiazine-4-
carboxylic acid (EPC), 10-phenoxazinepropionic acid (POP) and 10-
methylphenoxazine (described in WO 94/12621 ) and substitued syringates (C3-
C5 substitued alkyl syringates) and phenols. Sodium percarbonate or perborate
are preferred sources of hydrogen peroxide.
Said peroxidases are normally incorporated in the detergent composition at
levels from 0.0001 % to 2% of pure enzyme by weight of the detergent
composition.
Other preferred enzymes that can be included in the detergent compositions of
the present invention include lipases. Suitable lipase enzymes for detergent
usage include those produced by microorganisms of the Pseudomonas group,
such as Pseudomonas stutzeri ATCC 19.154, as disclosed in British Patent
1,372,034. Suitable lipases include those which show a positive immunological
cross-reaction with the antibody of the lipase, produced by the microorganism
Pseudomonas fluorescent IAM 1057. This lipase is available from Amano
Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name Lipase P
"Amano," hereinafter referred to as "Amano-P". Other suitable commercial
lipases include Amano-CES, lipases ex Chromobacter viscosum, e.g.
Chromobacter viscosum var. lipolyticum NRRLB 3673 from Toyo Jozo Co.,
Tagata, Japan; Chromobacter viscosum lipases from U.S. Biochemical Corp.,
U.S.A. and Disoynth Co., The Netherlands, and lipases ex Pseudomonas
gladioli. Especially suitable lipases are lipases such as M 1 LipaseR and
LipomaxR (Gist-Brocades) and LipolaseR and Lipolase UItraR(Novo) which have
found to be very effective when used in combination with the compositions of
the
present invention. Also suitables are the lipolytic enzymes described in EP
258
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068, WO 92/05249 and WO 95/22615 by Novo Nordisk and in WO 94/03578,
WO 95/35381 and WO 96/00292 by Unilever.
Also suitable are cutinases [EC 3.1.1.50] which can be considered as a special
kind of lipase, namely lipases which do not require interfacial activation.
Addition
of cutinases to detergent compositions have been described in e.g. WO-A-
88/09367 (Genencor); WO 90/09446 (Plant Genetic System) and WO 94/14963
and WO 94/14964 (Unilever).
The lipases and/or cutinases are normally incorporated in the detergent
composition at levels from 0.0001 % to 2% of pure enzyme by weight of the
detergent composition.
Amylases (a and/or f3) can be included for removal of carbohydrate-based
stains.
W094/02597, Novo Nordisk A/S published February 03, 1994, describes
detergent compositions which incorporate mutant amylases. See also
W095/10603, Novo Nordisk A/S, published April 20, 1995. Other amylases
known for use in detergent compositions include both a- and ~-amylases. a-
Amylases are known in the art and include those disclosed in US Pat. no.
5,003,257; EP 252,666; WO/91/00353; FR 2,676,456; EP 285,123; EP 525,610;
EP 368,341; and British Patent specification no. 1,296,839 (Novo). Other
suitable
amylases are stability-enhanced amylases described in W094/18314, published
August 18, 1994 and W096/05295, Genencor, published February 22, 1996 and
amylase variants having additional modification in the immediate parent
available
from Novo Nordisk A/S, disclosed in WO 95/10603, published April 95. Also
suitable are amylases described in EP 277 216, W095/26397 and W096/23873
(all by Novo Nordisk).
Examples of commercial a-amylases products are Purafect Ox Am~ from
Genencor and Termamyl~, Ban~ ,Fungamyl~ and Duramyl~, all available from
Novo Nordisk A/S Denmark. W095/26397 describes other suitable amylases : a-
amylases characterised by having a specific activity at least 25% higher than
the
specific activity of Termamyl~ at a temperature range of 25°C to
55°C and at a
pH value in the range of 8 to 10, measured by the Phadebas~ a-amylase activity
assay. Suitable are variants of the above enzymes, described in W096/23873
(Novo Nordisk). Other amylolytic enzymes with improved properties with respect
to the activity level and the combination of thermostability and a higher
activity
level are described in W095/35382.
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21
The amylolytic enzymes are incorporated in the detergent compositions of the
present invention a level of from 0.0001 % to 2%, preferably from 0.00018% to
0.06%, more preferably from 0.00024% to 0.048% pure enzyme by weight of the
composition.
The above-mentioned enzymes may be of any suitable origin, such as vegetable,
animal, bacterial, fungal and yeast origin. Origin can further be mesophilic
or
extremophilic (psychrophilic, psychrotrophic, thermophilic, barophilic,
alkalophilic,
acidophilic, halophilic, etc.). Purified or non-purified forms of these
enzymes may
be used. Nowadays, it is common practice to modify wild-type enzymes via
protein / genetic engineering techniques in order to optimise their
performance
efficiency in the detergent compositions of the invention. For example, the
variants may be designed such that the compatibility of the enzyme to commonly
encountered ingredients of such compositions is increased. Alternatively, the
variant may be designed such that the optimal pH, bleach or chelant stability,
catalytic activity and the like, of the enzyme variant is tailored to suit the
particular cleaning application.
In particular, attention should be focused on amino acids sensitive to
oxidation in
the case of bleach stability and on surface charges for the surfactant
compatibility. The isoelectric point of such enzymes may be modified by the
substitution of some charged amino acids, e.g. an increase in isoelectric
point
may help to improve compatibility with anionic surfactants. The stability of
the
enzymes may be further enhanced by the creation of e.g. additional salt
bridges
and enforcing calcium binding sites to increase chelant stability. Special
attention
must be paid to the cellulases as most of the cellulases have separate binding
domains (CBD). Properties of such enzymes can be altered by modifications in
these domains.
Said enzymes are normally incorporated in the detergent composition at levels
from 0.0001 % to 2% of pure enzyme by weight of the detergent composition. The
enzymes can be added as separate single ingredients (prills, granulates,
stabilized liquids, etc. containing one enzyme ) or as mixtures of two or more
enzymes (e.g. cogranulates ).
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22
Other suitable detergent ingredients that can be added are enzyme oxidation
scavengers which are described in co-pending European Patent application
92870018.6 filed on January 31, 1992. Examples of such enzyme oxidation
scavengers are ethoxylated tetraethylene polyamines.
A range of enzyme materials and means for their incorporation into synthetic
detergent compositions is also disclosed in WO 9307263 A and WO 9307260 A
to Genencor International, WO 8908694 A to Novo, and U.S. 3,553,139, January
5, 1971 to McCarty et al. Enzymes are further disclosed in U.S. 4,101,457,
Place
et al, July 18, 1978, and in U.S. 4,507,219, Hughes, March 26, 1985. Enzyme
materials useful for liquid detergent formulations, and their incorporation
into
such formulations, are disclosed in U.S. 4,261,868, Hora et al, April 14,
1981.
Enzymes for use in detergents can be stabilised by various techniques. Enzyme
stabilisation techniques are disclosed and exemplified in U.S. 3,600,319,
August
17, 1971, Gedge et al, EP 199,405 and EP 200,586, October 29, 1986,
Venegas. Enzyme stabilisation systems are also described, for example, in U.S.
3,519,570. A useful Bacillus, sp. AC13 giving proteases, xylanases and
cellulases, is described in WO 9401532 A to Novo.bacterial and fungal
cellulases
such as Carezyme and Celluzyme (Novo Nordisk A/S); peroxidases; lipases
such as Amano-P (Amano Pharmaceutical Co.), M1 LipaseR and LipomaxR (Gist-
Brocades) and LipolaseR and Lipolase UItraR (Novo); cutinases; proteases such
as EsperaseR, AlcalaseR, DurazymR and SavinaseR (Novo) and MaxataseR,
MaxacalR, ProperaseR and MaxapemR (Gist-Brocades); and a and ~i amylases
such as Purafect Ox AmR (Genencor) and TermamylR, Bang, FungamylR,
DuramylR, and NatalaseR (Novo); and mixtures thereof. Enzymes are preferably
added herein as prills, granulates, or cogranulates at levels typically in the
range
from about 0.0001 % to about 2% pure enzyme by weight of composition.
Bleaching agents suitable herein include chlorine and oxygen bleaches,
especially inorganic perhydrate salts such as sodium perborate mono-and
tetrahydrates and sodium percarbonate optionally coated to provide controlled
rate of release (see, for example, GB-A-1466799 on sulfate/carbonate
coatings),
preformed organic peroxyacids and mixtures thereof with organic peroxyacid
bleach precursors and/or transition metal-containing bleach catalysts
(especially
manganese or cobalt). Inorganic perhydrate salts are typically incorporated at
levels in the range from about 1 % to about 40% by weight, preferably from
about
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23
2% to about 30% by weight and more preferably from abut 5% to about 25% by
weight of composition. Peroxyacid bleach precursors preferred for use herein
include precursors of perbenzoic acid and substituted perbenzoic acid;
cationic
peroxyacid precursors; peracetic acid precursors such as TAED, sodium
acetoxybenzene sulfonate and pentaacetylglucose; pernonanoic acid precursors
such as sodium 3,5,5-trimethylhexanoyloxybenzene sulfonate (iso-NOBS) and
sodium nonanoyloxybenzene sulfonate (NOBS); Phenolsulfonate ester of N-
nonanoyl-6-aminocaproic acid (NACA-OBS, described in W094/28106), which
are perhydrolyzed to form a peracid as the active bleaching species, leading
to
improved bleaching effect, amide substituted alkyl peroxyacid precursors (EP-A-
0170386); and benzoxazin peroxyacid precursors (EP-A-0332294 and EP-A-
0482807). Bleach precursors are typically incorporated at levels in the range
from about 0.5% to about 25%, preferably from about 1 % to about 10% by
weight of composition while the preformed organic peroxyacids themselves are
typically incorporated at levels in the range from 0.5% to 25% by weight, more
preferably from 1 % to 10% by weight of composition. Bleach catalysts
preferred
for use herein include the manganese triazacyclononane and related complexes
(US-A-4246612, US-A-5227084); Co, Cu, Mn and Fe bispyridylamine and related
complexes (US-A-5114611); and pentamine acetate cobalt(III) and related
complexes(US-A-4810410).
Other suitable components herein include organic polymers having dispersant,
anti-redeposition, soil release or other detergency properties invention in
levels of
from about 0.1 % to about 30%, preferably from about 0.5% to about 15%, most
preferably from about 1 % to about 10% by weight of composition. Preferred
anti-
redeposition polymers herein include acrylic acid containing polymers such as
Sokalan PA30, PA20, PA15, PA10 and Sokalan CP10 (BASF GmbH), Acusol
45N, 480N, 460N (Rohm and Haas), acrylic acid/maleic acid copolymers such as
Sokalan CP5 and acrylic/methacrylic copolymers. Preferred soil release
polymers herein include alkyl and hydroxyalkyl celluloses (US-A-4,000,093),
polyoxyethylenes, polyoxypropylenes and copolymers thereof, and nonionic and
anionic polymers based on terephthalate esters of ethylene glycol, propylene
glycol and mixtures thereof.
Heavy metal seguestrants and crystal growth inhibitors are suitable for use
herein in levels generally from about 0.005% to about 20%, preferably from
about
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24
0.1 % to about 10%, more preferably from about 0.25% to about 7.5% and most
preferably from about 0.5% to about 5% by weight of composition, for example
diethylenetriamine penta (methylene phosphonate), ethylenediamine
tetra(methylene phosphonate) hexamethylenediamine tetra(methylene
phosphonate), ethylene diphosphonate, hydroxy-ethylene-1,1-diphosphonate,
nitrilotriacetate, ethylenediaminotetracetate, ethylenediamine-N,N'-
disuccinate in
their salt and free acid forms.
The compositions herein, especially for use in dishwashing, can contain a
corrosion inhibitor such as organic silver coating agents in levels of from
about
0.05% to about 10%, preferably from about 0.1 % to about 5% by weight of
composition (especially paraffins such as Winog 70 sold by Wintershall,
Salzbergen, Germany), nitrogen-containing corrosion inhibitor compounds (for
example benzotriazole and benzimadazole - see GB-A-1137741 ) and Mn(II)
compounds, particularly Mn(II) salts of organic ligands in levels of from
about
0.005% to about 5%, preferably from about 0.01 % to about 1 %, more preferably
from about 0.02% to about 0.4% by weight of the composition.
Other suitable components herein include colourants, water-soluble bismuth
compounds such as bismuth acetate and bismuth citrate at levels of from about
0.01 % to about 5%, enzyme stabilizers such as calcium ion, boric acid,
propylene glycol and chlorine bleach scavengers at levels of from about 0.01 %
to
about 6%, lime soap dispersants (see WO-A-93/08877), suds suppressors (see
WO-93/08876 and EP-A-0705324), polymeric dye transfer inhibiting agents,
optical brighteners, perfumes, fillers and clay and cationic fabric softeners.
The detergent tablets herein are preferably formulated to have a not unduly
high
pH, preferably a pH in 1 % solution in distilled water of from about 8.0 to
about
12.5, more preferably from about 9.0 to about 11.8, most preferably from about
9.5 to about 11.5.
Preferably, the tablet will be the tablets described in the co-pending
European
application No. 9815525.2 filed 17 July 1998.
Such tablets are multi-phase detergent tablets for use in a washing machine,
the
tablet comprising:
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WO 00/42148 PCT/US99/00800
a) a slower dissolving phase in the form of a shaped body having at least one
mould therein (Section 1); and
b) a second rapid dissolving phase in the form of a particulate solid
compressed
within said mould (Section 2), comprising the pectate lyase enzyme of the
present invention.
In preferred embodiments, the first phase is a compressed shaped body
prepared at an applied compression pressure of at least about 350 kg/cm2 (3.43
kN/cm2), preferably from about 400 to about 2000, especially from about 600 to
about 1200 kg/cm2 (compression pressure herein is the applied force divided by
the cross-sectional area of the tablet in a plane transverse to the applied
force -
in effect, the transverse cross-sectional area of the die of the rotary
press). It is
also preferred that the particulate solid of the second phase (which
terminology is
intended to include the possibility of multiple 'second' phases, sometimes
referred to herein as 'optional subsequent phases') be compressed into said
mould at an applied compression pressure less than that applied to the first
phase and preferably at a compression pressure of less than about 350 kg/cm2,
preferably in the range from about 40 kg/cm2 to about 300 kg/cm2 and more
preferably from about 70 to about 270 kg/cm2, such tablets being preferred
herein from the viewpoint of providing optimum tablet integrity and strength
(measured for example by the Child Bite Strength [CBS] test) and product
dissolution characteristics. The tablets of the invention preferably have a
CBS of
at least 10kg, preferably greater than 12kg, more preferably greater than
14kg,
CBS being measured per the US Consumer Product Safety Commission Test
Specification. Also, the compression pressures applied to the first and second
phases will generally be in a ratio of at least about 2:1, preferably at least
about
4:1.
Thus, according to a further aspect of the invention, there is provided a
multi-
phase detergent tablet for use in a washing machine, the tablet comprising:
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WO 00/42148 PCT/US99/00800
26
a) a first slow dissolving phase in the form of a compressed shaped body
having
at least one mould therein, the shaped body being prepared at a compression
pressure of at least about 350 kg/cm2; and
b) a second phase in the form of a particulate solid compressed within said
mould, the second phase being compressed at a pressure of less than about 350
kg/cm2, comprising the pectate lyase enzyme of the present invention
In other preferred embodiments, the second phase is in the form of a
compressed or shaped body adhesively contained, for example by physical or
chemical adhesion, within the at least one mould of the first body. It is also
preferred that the first and second phases are in a relatively high weight
ratio to
one another, for example at least about 6:1, preferably at least about 10:1;
also
that the tablet composition contain one or more detergent actives (for example
enzymes, bleaches, bleach activators, bleach catalysts, surfactants, chelating
agents etc) which is predominantly concentrated in the second phase, for
example, at least about 50%, preferably at least about 60%, especially about
80% by weight of the active (based on the total weight of the active in
tablet) is in
the second phase of the tablet. In this case specifically the preferred active
is the
pectate lyase enzyme. Again, such compositions are optimum for tablet
strength,
dissolution, cleaning, and pH regulation characteristics providing, for
example,
tablet compositions capable of dissolving in the wash liquor so as to deliver
at
least 50%, preferably at least 60%, and more preferably at least 80% by weight
of the pectate lyase to the wash liquor within 10, 5, 4 or even 3 minutes of
the
start of the wash process.
Thus, according to another aspect of the invention, there is provided a multi-
phase detergent tablet for use in a washing machine, the tablet comprising:
a) slow dissolving first phase in the form of a shaped body having at least
one
mould therein (Section 1 ), and
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WO 00/42148 PCT/LJS99/00800
27
b) a rapid dissolving second phase containing pectate lyase and in the form of
a
particulate solid compressed within said mould (Section 2),
and wherein section 2 of the tablet comprises at 70%, preferably at least 85%,
more preferably at least 95% by weight of the pectate lyase which is delivered
to
the wash within the first 10 minutes, preferably within the first 5 minutes,
and
more preferably within the first 3 minutes of the wash process.
Also suitable are the following tablets, specifically designed for laundry
purposes
Tablet Manufacture
Detergent tablets can be prepared simply by mixing the solid ingredients
together
and compressing the mixture in a conventional tablet press as used, for
example,
in the pharmaceutical industry. Preferably the principal ingredients, in
particular
gelling surfactants, are used in particulate form. Any liquid ingredients, for
example surfactant or suds suppressor, can be incorporated in a conventional
manner into the solid particulate ingredients.
In particular for laundry tablets, the ingredients such as builder and
surfactant
can be spray-dried in a conventional manner and then compacted at a suitable
pressure. Preferably, the tablets according to the invention are compressed
using
a force of less than 100000N, more preferably of less than 50000N, even more
preferably of less than 5000N and most preferably of less than 3000 N. Indeed,
the most preferred embodiment is a tablet suitable for laundry compressed
using
a force of less than 2500N, but tablets for auto dish washing may also be
considered for example, whereby such auto dish washing tablets are usually
more compressed than laundry tablets.
The particulate material used for making the tablet of this invention can be
made
by any particulation or granulation process. An example of such a process is
spray drying (in a co-current or counter current spray drying tower) which
typically gives low bulk densities 600g/I or lower. Particulate materials of
higher
density can be prepared by granulation and densification in a high shear batch
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28
mixer/granulator or by a continuous granulation and densification process
(e.g.
using Lodige~ CB and/or Lodige~ KM mixers). Other suitable processes include
fluid bed processes, compaction processes (e.g. roll compaction), extrusion,
as
well as any particulate material made by any chemical process like
flocculation,
crystallisation sentering, etc. Individual particles can also be any other
particle,
granule, sphere or grain.
The components of the particulate material may be mixed together by any
conventional means. Batch is suitable in, for example, a concrete mixer, Nauta
mixer, ribbon mixer or any other. Alternatively the mixing process may be
carried
out continuously by metering each component by weight on to a moving belt, and
blending them in one or more drums) or mixer(s). Non-gelling binder can be
sprayed on to the mix of some, or all of, the components of the particulate
material. Other liquid ingredients may also be sprayed on to the mix of
components either separately or premixed. For example perfume and slurries of
optical brighteners may be sprayed. A finely divided flow aid (dusting agent
such
as zeolites, carbonates, silicas) can be added to the particulate material
after
spraying the binder, preferably towards the end of the process, to make the
mix
less sticky.
The tablets may be manufactured by using any compacting process, such as
tabletting, briquetting, or extrusion, preferably tabletting. Suitable
equipment
includes a standard single stroke or a rotary press (such as Courtoy~, Korch~,
Manesty~, or Bonals~). The tablets prepared according to this invention
preferably have a diameter of between 20mm and 60mm, preferably of at least
35 and up to 55 mm, and a weight between 25 and 100 g. The ratio of height to
diameter (or width) of the tablets is preferably greater than 1:3, more
preferably
greater than 1:2. The compaction pressure used for preparing these tablets
need
not exceed 100000 kN/m2, preferably not exceed 30000 kN/m2, more preferably
not exceed 5000 kN/m2, even more preferably not exceed 3000kN/m2 and most
preferably not exceed 1000kN/m2. In a preferred embodiment according to the
invention, the tablet has a density of at least 0.9 g/cc, more preferably of
at least
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29
1.0 g/cc, and preferably of less than 2.0 g/cc, more preferably of less than
1.5
g/cc, even more preferably of less than 1.25 g/cc and most preferably of less
than 1.1 g/cc.
Multi layered tablets are typically formed in rotating presses by placing the
matrices of each layer, one after the other in matrix force feeding flasks. As
the
process continues, the matrix layers are then pressed together in the pre-
compression and compression stages stations to form the multilayer layer
tablet.
With some rotating presses it is also possible to compress the first feed
layer
before compressing the whole tablet.
Hydrotro~e compound
In a preferred embodiment of the invention, a highly soluble compound having a
cohesive effect is integrated to the tablet of the invention, whereby this
compound is also a hydrotrope compound. Such hydrotrope compound may be
generally used to favour surfactant dissolution by avoiding gelling, so that
they
may be for example advantageously comprised in a softer faster dissolving
layer
which also contains the pectate lyase. A specific compound is defined as being
hydrotrope as follows (see S.E. Friberg and M. Chiu, J. Dispersion Science and
Technology, 9(5&6), pages 443 to 457, (1988-1989)):
1. A solution is prepared comprising 25% by weight of the specific compound
and 75% by weight of water.
2. Octanoic Acid is thereafter added to the solution in a proportion of 1.6
times
the weight of the specific compound in solution, the solution being at a
temperature of 20°Celsius. The solution is mixed in a Sotax beaker with
a
stirrer with a marine propeller, the propeller being situated at about 5mm
above the bottom of the beaker, the mixer being set at a rotation speed of
200 rounds per minute.
3. The specific compound is hydrotrope if the the Octanoic Acid is completely
solubilised, i.e . if the solution comprises only one phase, the phase being a
liquid phase.
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It should be noted that in a preferred embodiment of the invention, the
hydrotrope compound is a flowable material made of solid particles at
operating
conditions between 15 and 60° Celsius.
Hydrotrope compounds include the compounds listed thereafter:
A list of commercial hydrotropes could be found in McCutcheon's Emulsifiers
and
Detergents published by the McCutcheon division of Manufacturing
Confectioners Company. Compounds of interest also include:
1. Nonionic hydrotrope with the following structure:
R - O - (CH2CH20)x( CH -CH20)yH
CH3
where R is a C8-C10 alkyl chain, x ranges from 1 to 15, y from 3 to 10.
2. Anionic hydrotropes such as alkali metal aryl sulfonates. This includes
alkali
metal salts of benzoic acid, salicylic acid, bezenesulfonic acid and its many
derivatives, naphthoic acid and various hydroaromatic acids. Examples of these
are sodium, potassium and ammonium benzene sulfonate salts derived from
toluene sulfonic acid, xylene sulfonic acid, cumene sulfonic acid, tetralin
sulfonic
acid, naphtalene sulfonic acid, methyl- naphtalene sulfonic acid, dimethyl
naphtalene sulfonic acid, trimethyl naphtalene sulfonic acid=
Other examples include salts of dialkyl benzene sulfonic acid such as salts of
di-
isopropyl benzene sulfonic acid, ethyl methyl benzene sulfonic acid, alkyl
benzene sulfonic acid with an alkyl chain length with 3 to 10, (pref. 4 to 9),
linear
or branched alkyl sulfonates with an alkyl chain with 1 to 18 carbons.
3. Solvent hydrotropes such as alkoxylated glycerines and alkoxylated
glycerides, esters slakoxylated glycerines, alkoxylated fatty acids, esters of
glycerin, polyglycerol esters. Preferred alkoxylated glycerines have the
following
structure:
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31
R
CH2-O(-CH2CH-O-)mH
I R
CH -O -CH CH-O- H
2 ( 2 )m
R
CH2-O(- H2CH-O-)nH
where I, m and n are each a number from 0 to about 20, with I+m+n = from about
2 to about 60, preferably from about 10 to about 45 and R represents H, CH3 or
CZHs
Preferred alkoxylated glycerides have the following struture
H2 -R~
H _R2 R3
H C-O- GH CH-O -H
2 ( 2 )
where R1 and R2 are each C~COO or -(CH2CHR3 O),-H where R3 = H, CH3 or
CZHS and I is a number from 1 to about 60, n is a number from about 6 to about
24.
4. Polymeric hydrotropes such as those described in EP636687:
R R~
-(CH -C)x - ( H2-C)y-
E RZ
where E is a hydrophilic functional group,
R is H or a C1-C10 alkyl group or is a hydrophilic functional group;
R1 is H a lower alkyl group or an aromatic group,
R2 is H or a cyclic alkyl or aromatic group.
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32
The polymer typically has a molecular weight of between about 1000 and
1000000.
5. Hydrotrope of unusual structure such as 5-carboxy-4-hexyl-2-cyclohexene-1-
yl
octanoic acid (Diacid~)
Use of such compound in the invention would further increase the dissolution
rate of the tablet, as a hydrotrope compound facilitates dissolution of
surfactants,
for example. Such a compound could be formed from a mixture or from a single
compound.
Coating
In another embodiment of the present invention, the solidity of the tablet
according to the invention may be further improved by making a coated tablet,
the coating covering a non-coated tablet according to the invention and
containing the pectate lyase enzyme, thereby further improving the mechanical
characteristics of the tablet while allowing rapid dissolution of the pectate
lyase
enzyme.
This very advantageously applies to multi-layer tablets according to the
invention,
whereby the dissolution characteristics of the outside layer can be tailored
to
allow fast release of the coating ingredients, thus combining the advantage of
the
coating with the advantage of time release.
In one embodiment of the present invention, the tablets may be coated so that
the tablet does not absorb moisture, or absorbs moisture at only a very slow
rate.
The coating is also strong so that moderate mechanical shocks to which the
tablets are subjected during handling, packing and shipping result in no more
than very low levels of breakage or attrition. Finally the coating is
preferably
brittle so that the tablet breaks up when subjected to stronger mechanical
shock.
Furthermore it is advantageous if the coating material is dissolved under
alkaline
conditions, or is readily emulsified by surfactants to allow release of the
Pectate
Lyase. This also contributes to avoiding the problem of visible residue in the
window of a front-loading washing machine during the wash cycle, and also
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33
avoids deposition of undissolved particles or lumps of coating material on the
laundry load.
Water solubility is measured following the test protocol of ASTM E1148-87
entitled, "Standard Test Method for Measurements of Aqueous Solubility".
Suitable coating materials that can be used in combination with pectate lyase
are
dicarboxylic acids. Particularly suitable dicarboxylic acids are selected from
the
group consisting of oxalic acid, malonic acid, succinic acid, glutaric acid,
adipic
acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, undecanedioic
acid,
dodecanedioic acid, tridecanedioic acid and mixtures thereof.
The coating material has a melting point preferably of from 40 °C to
200 °C.
The coating can be applied in a number of ways. However, the preferred method
when pectate lyase is contained in the coating is to coat with a solution of
the
material. In this method, the coating is applied as a solution, the solvent
being
dried to leave a coherent coating. The substantially insoluble material can be
applied to the tablet by, for example, spraying or dipping. Clearly
substantially
insoluble materials having a melting point below 40 °C are not
sufficiently solid at
ambient temperatures and it has been found that materials having a melting
point
above about 200 °C are not practicable to use. Preferably, the
materials melt in
the range from 60 °C to 160 °C, more preferably from 70
°C to 120 °C.
By "melting point" is meant the temperature at which the material when heated
slowly in, for example, a capillary tube becomes a clear liquid.
A coating of any desired thickness can be applied according to the present
invention. For most purposes, the coating forms from 1 % to 10%, preferably
from 1.5% to 5%, of the tablet weight.
The tablet coatings of the present invention are very hard, provide extra
strength
to the tablet, and allow for early release of the pectate lyase.
In a preferred embodiment of the present invention the fracture of the coating
in
the wash is improved by adding a disintegrant in the coating. This
disintegrant
will swell once in contact with water and break the coating in small pieces.
This
will improve the dissolution of the coating in the wash solution. The
disintegrant is
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34
suspended in the coating melt at a level of up to 30%, preferably between 5%
and 20%, most preferably between 5 and 10% by weight. Possible disintegrants
are described in Handbook of Pharmaceutical Excipients (1986). Examples of
suitable disintegrants include starch: natural, modified or pregelatinized
starch,
sodium starch gluconate; gum: agar gum, guar gum, locust bean gum, karaya
gum, pectin gum, tragacanth gum; croscarmylose Sodium, crospovidone,
cellulose, carboxymethyl cellulose, algenic acid and its salts including
sodium
alginate, silicone dioxide, clay, polyvinylpyrrolidone, soy polysacharides,
ion
exchange resins and mixtures thereof.
Tensile Strength
For the purpose of measuring tensile strength of a layer, the layer may be
considered as a tablet itself.
Depending on the composition of the starting material, and the shape of the
tablets, the used compacting force may be adjusted to not affect the tensile
strength, and the disintegration time in the washing machine. This process may
be used to prepare homogenous or layered tablets of any size or shape.
For a cylindrical tablet, the tensile strength corresponds to the diametrical
fracture stress (DFS) which is a way to express the strength of a tablet or
layer,
and is determined by the following equation
Tensile strength = 2 F/ ~Dt
Where F is the maximum force (Newton) to cause tensile failure (fracture)
measured by a VK 200 tablet hardness tester supplied by Van Kell industries,
Inc. D is the diameter of the tablet or layer, and t the thickness of the
tablet or
layer. For a non round tablet, ~D may simply be replaced by the perimeter of
the
tablet.
(Method Pharmaceutical Dosage Forms : Tablets Volume 2 Page 213 to 217).
A tablet having a diametral fracture stress of less than 20 kPa is considered
to be
fragile and is likely to result in some broken tablets being delivered to the
consumer. A diametral fracture stress of at least 25 kPa is preferred.
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This applies similarly to non cylindrical tablets, to define the tensile
strength,
whereby the cross section normal to the height of the tablet is non round, and
whereby the force is applied along a direction perpendicular to the direction
of
the height of the tablet and normal to the side of the tablet, the side being
perpendicular to the non round cross section.
Effervescent
In another preferred embodiment of the present invention the tablets further
comprises an effervescent.
Effervescency as defined herein means the evolution of bubbles of gas from a
liquid, as the result of a chemical reaction between a soluble acid source and
an
alkali metal carbonate, to produce carbon dioxide gas,
i.e. CgH8O7 + 3NaHC03 -~ Na3CgH507 + 3C02 T + 3H20
Further examples of acid and carbonate sources and other effervescent systems
may be found in : (Pharmaceutical Dosage Forms : Tablets Volume 1 Page 287
to 291 ).
An effervescent may be added to the tablet mix in addition to the detergent
ingredients. The addition of this effervescent to the detergent tablet
improves the
disintegration time of the tablet. The amount will preferably be between 5 and
20
and most preferably between 10 and 20% by weight of the tablet. Preferably
the effervescent should be added as an agglomerate of the different particles
or
as a compact, and not as separated particles.
Due to the gas created by the effervescency in the tablet, the tablet can have
a
higher D.F.S. and still have the same disintegration time as a tablet without
effervescency. When the D.F.S. of the tablet with effervescency is kept the
same
as a tablet without, the disintegration of the tablet with effervescency will
be
faster.
Further dissolution aid could be provided by using compounds such as sodium
acetate or urea. A list of suitable dissolution aid may also be found in
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36
Pharmaceutical Dosage Forms: Tablets, Volume 1, Second edition, Edited by
H.A. Lieberman et all, ISBN 0-8247-8044-2.
Other components
Surfactant are comprised in the tablet according to the invention. Suitable
surfactants are described herein above. Also suitable for the present tablet
are
builders, bleaching agents, enzymes and enzymes herein above described
Non ctelling binders
Non gelling binders can be integrated to the particles forming the tablet in
order
to further facilitate dissolution.
If non gelling binders are used, suitable non-gelling binders include
synthetic
organic polymers such as polyethylene glycols, polyvinylpyrrolidones,
polyacrylates and water-soluble acrylate copolymers. The handbook of
Pharmaceutical Excipients second edition, has the following binders
classification: Acacia, Alginic Acid, Carbomer, Carboxymethylcellulose sodium,
Dextrin, Ethylcellulose, Gelatin, Guar gum, Hydrogenated vegetable oil type I,
Hydroxyethyl cellulose, Hydroxypropyl methylcellulose, Liquid glucose,
Magnesium aluminum silicate, Maltodextrin, Methylcellulose, polymethacrylates,
povidone, sodium alginate, starch and zein. Most preferable binders also have
an
active cleaning function in the laundry wash such as cationic polymers, i.e.
ethoxylated hexamethylene diamine quaternary compounds, bishexamethylene
triamines, or others such as pentaamines, ethoxylated polyethylene amines,
malefic acrylic polymers.
Non-gelling binder materials are preferably sprayed on and hence have an
appropriate melting point temperature below 90°C, preferably below
70°C and
even more preferably below 50°C so as not to damage or degrade the
other
active ingredients in the matrix. Most preferred are non-aqueous liquid
binders
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37
(i.e. not in aqueous solution) which may be sprayed in molten form. However,
they may also be solid binders incorporated into the matrix by dry addition
but
which have binding properties within the tablet.
Non-gelling binder materials are preferably used in an amount within the range
from 0.1 to 15% of the composition, more preferably below 5% and especially if
it
is a non laundry active material below 2% by weight of the tablet.
It is preferred that gelling binders, such as nonionic surfactants are avoided
in
their liquid or molten form. Nonionic surfactants and other gelling binders
are not
excluded from the compositions, but it is preferred that they be processed
into
the detergent tablets as components of particulate materials, and not as
liquids.
The Pectate Lyase enzyme
An essential element of the detergent tablets of the present invention is a
pectate
lyase enzyme.
Pectate lyase is classified within the classification of enzymes provided by
the
Enzyme Nomenclature (1992) as EC 4.2.2.2. Said enzyme is known to split the
a-1,4,glucoside bond of galacturonic acid found in pectin substances, creating
a
double bond between C4 and C5 and is substantially free for other pectin
degrading activities, i.e having less than 25%, preferably less than 15%, more
preferably less than 5% by weight of the enzyme compound of other pectin
degrading enzyme activities.
Pectate lyases have been cloned from different bacterial genera such as
Erwinia,
Pseudomonas, Klebsiella and Xanthomonas, Streptomyces, Penicillium,
Baceriodes, Thermomonospora, Fusarium, and Aspergillus. Also from Bacillus
subtilis (Nasser et al. (1993) FEBS 335:319-326) and Bacillus sp. YA-14 (Kim
et
al. (1994) Biosci. Biotech. Biochem. 58:947-949) cloning of a pectate lyase
has
been described. Purification of pectate lyases with maximum activity in the pH
range of 8-10 produced by Bacillus pumilus (Dave and Vaughn (1971) J.
Bacteriol. 108:166-174), 8. polymyxa (Nagel and Vaughn (1961) Arch. Biochem.
Biophys. 93:344-352), 8. stearothermophilus (Karbassi and Vaughn (1980) Can.
J. Microbiol. 26:377-384), Bacillus sp. (Hasegawa and Nagel (1966) J. Food
Sci.
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38
31:838-845) and Bacillus sp. RK9 (Kelly and Fogarty (1978) Can. J. Microbiol.
24:1164-1172) has been reported. WO 98/45393 discloses detergent
compositions containing protopectinase with remarkable detergency against
muddy soils.
Further suitable pectate lyases for use in the present invention are the
protopectinases having an optimum reaction pH of 7.0 or higher when
polygalacturonic acid is used as a substrate such as described in W098/45393
and the pectic acid lyase having the amino acid sequence SEQ no 1 of EP 870
843 or having such amino acid sequence with one or more amino acid being
deleted, added or substituted.
Preferred are the pectate lyase enzymes described in the international co-
pending application PCT/DK98/00515, first filed in Denmark on November 24,
1997
- A pectate lyase comprising a first amino acid sequence consisting of seven
(7)
amino acid residues having the following sequence: Asn Leu Asn Ser Arg Val
Pro (NLNSRVP);
- A pectate lyase which is
i) a polypeptide produced by Bacillus agaradhaerens, NCIMB 40482 or
DSM 8721, or by a Bacillus species having a 16S rDNA sequence
homology to Bacillus agaradhaerens, DSM 8721, of at least 99%, or
ii) a polypeptide comprising an amino acid sequence as shown in positions
27-359 of SEQ ID N0:2 of PCT/DK98/00515, or
iii) an analogue of the polypeptide defined in i) or ii) which is at least 45%
homologous with said polypeptide, or
iv) is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the arginine in position 240, and
optionally also the arginine in position 245, is conserved and the derived
polypeptide is at least 42% homologous with said polypeptide, or
v) is immunologically reactive with a polycional antibody raised against
said polypeptide in purified form;
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39
- A pectate lyase which is
i) a polypeptide produced by Bacillus licheniformis, ATCC 14580, or by a
Bacillus species having a 16S rDNA sequence homology to Bacillus
licheniformis, ATCC 14580, of at least 99%, or
ii) a polypeptide comprising an amino acid sequence as shown in positions
28-341 of SEQ ID N0:4 of PCT/DK98100515, or
iii) an analogue of the polypeptide defined in i) or ii) which is at least 45%
homologous with said polypeptide, or
iv) is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the arginine in position 233, and
optionally also the arginine in position 238, is conserved and the derived
polypeptide is at least 42% homologous with said polypeptide, or
v) is immunologically reactive with a polyclonal antibody raised against
said polypeptide in purified form;
- A pectate lyase which is
i) a polypeptide produced by a Bacillus species having the 16S rDNA
sequence of SEQ ID N0:14 or by a Bacillus species having a 16S rDNA
sequence homology to SEQ ID N0:14 higher than 97.3%; or
ii) a polypeptide comprising an amino acid sequence as shown in positions
181-509 of SEQ ID N0:6, or
iii) an analogue of the polypeptide defined in i) which is at least 50%
homologous with said polypeptide, or
iv) is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the arginine in position 390, and
optionally also the arginine in position 395, is conserved and the derived
polypeptide is at least 44% homologous with said polypeptide, or
v) is immunologically reactive with a polyclonal antibody raised against
said polypeptide in purified form,
- A pectate lyase which is
i) a polypeptide produced by the species Bacillus halodurans, or
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ii) a polypeptide comprising an amino acid sequence as shown in positions
42-348 of SEQ ID N0:8 of PCT/DK98/00515, or
iii) an analogue of the polypeptide defined in i) or ii) which is at least 45%
homologous with said polypeptide, or
iv) is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the arginine in position 240, and
optionally also the arginine in position 245, is conserved and the derived
polypeptide is at least 40% homologous with said polypeptide, or
v) is immunologically reactive with a polyclonal antibody raised against
said polypeptide in purified form,
- A pectate lyase which is
i) a polypeptide produced by a Bacillus species having the 16S rDNA
sequence of SEQ ID N0:13 PCT/DK98/00515or by a Bacillus species
having a 16S rDNA sequence homology to SEQ ID N0:13 of
PCT/DK98/00515higher than 98.1 %; or
ii) a polypeptide comprising an amino acid sequence as shown in positions
25-335 of SEQ ID N0:10 of PCT/DK98/00515, or
iii) an analogue of the polypeptide defined in i) or which is at least 45%
homologous with said polypeptide, or
iv) is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the arginine in position 227, and
optionally also the arginine in position 232, is conserved and the derived
polypeptide is at least 41 % homologous with said polypeptide, or
v) is immunologically reactive with a polyclonal antibody raised against
said polypeptide in purified form.
Similarly preferred is the pectate lyase enzyme described in the international
co-
pending application PCT/DK98/00514, first filed in Denmark on November 24,
1997 and which is
~> a polypeptide produced by Bacillus licheniformis, ATCC 14580, or
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41
a polypeptide comprising an amino acid sequence as shown in
positions 28-221 of SEQ ID N0:4 of PCT/DK98/00514, or
an analogue of the polypeptide defined in i) or ii) which is at least
60% homologous with said polypeptide, or
is derived from said polypeptide by substitution, deletion or addition of
one or several amino acids, provided that the lysines in positions 133
and 155 and the arginine in position 158 are conserved and the
derived polypeptide is at least 66% homologous with positions 60-158
of SEQ ID N0:4 of PCT/DK98/00514, or
~> is immunologically reactive with a polyclonal antibody raised against
said polypeptide in purified form.
More preferred pectate lyases for the purpose of the present invention are
those
having optimum activity at pH's >7.0 and derived from Streptomyces fradiae,
Streptomyces nitrosporeus, Erwinia carotovora, Bacillus spheroides,
Thermomonospora fusca, Pseudomonas solanacearum, Bacteroides
thetaiotaomicron, Fusarium solani, Xanthomonas campestris, Bacillus
agaradhaerens, and/or Bacillus licheniformis.
Most preferred pectate lyase for the purpose of the present invention is the
Pectate lyase from Bacillus agaradhaerens, NCIMB 40482 or DSM 8721.
The pectate lyase is incorporated into the tablet of the invention preferably
at a
level of from 0.0001 % to 2%, more preferably from 0.0005% to 0.1 %, most
preferred from 0.001% to 0.02% pure enzyme by weight of the composition.
Preferably more than 70%, more preferably more than 85%, most preferably
more than 95% of the total amount of the pectate lyase enzyme will be
comprised in section 2 of the detergent tablet of the present invention.
The pectate lyase of the invention, in addition to the enzyme core comprising
the
catalytically domain, may also contain a cellulose binding domain (CBD), the
cellulose binding domain and enzyme core (the catalytically active domain) of
the
enzyme being operably linked. The cellulose binding domain (CBD) may exist as
an integral part of the encoded enzyme, or a CBD from another origin may be
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42
introduced into the enzyme thus creating an enzyme hybrid. In this context,
the
term "cellulose-binding domain" is intended to be understood as defined by
Peter
Tomme et al. "Cellulose-Binding Domains: Classification and Properties" in
"Enzymatic Degradation of Insoluble Carbohydrates", John N. Saddler and
Michael H. Penner (Eds.), ACS Symposium Series, No. 618, 1996. This definition
classifies more than 120 cellulose- binding domains into 10 families (I-X),
and
demonstrates that CBDs are found in various enzymes such as cellulases,
xylanases, mannanases, arabinofuranosidases, acetyl esterases and chitinases.
CBDs have also been found in algae, e.g. the red alga Porphyra purpurea as a
non-hydrolytic polysaccharide-binding protein, see Tomme et al., op.cit.
However, most of the CBDs are from cellulases and xylanases, CBDs are found
at the N and C termini of proteins or are internal. Enzyme hybrids are known
in
the art, see e.g. WO 90/00609 and WO 95/16782, and may be prepared by
transforming into a host cell a DNA construct comprising at least a fragment
of
DNA encoding the cellulose- binding domain ligated, with or without a linker,
to a
DNA sequence encoding the pectate lyase enzyme and growing the host cell to
express the fused gene. Enzyme hybrids may be described by the following
formula:
CBD - MR - X
wherein CBD is the N-terminal or the C-terminal region of an amino acid
sequence corresponding to at least the cellulose binding domain; MR is the
middle region (the linker), and may be a bond, or a short linking group
preferably
of from about 2 to about 100 carbon atoms, more preferably of from 2 to 40
carbon atoms; or is preferably from about 2 to about 100 amino acids, more
preferably of from 2 to 40 amino acids; and X is an N-terminal or C-terminal
region of the pectate lyase of the invention.
The above-mentioned enzymes may be of any suitable origin, such as vegetable,
animal, bacterial, fungal and yeast origin. Origin can further be mesophilic
or
extremophilic (psychrophilic, psychrotrophic, thermophilic, barophilic,
alkalophilic,
acidophilic, halophilic, etc.). Purified or non-purified forms of these
enzymes may
be used. Nowadays, it is common practice to modify wild-type enzymes via
protein / genetic engineering techniques in order to optimise their
performance
efficiency in the detergent compositions of the invention. For example, the
variants may be designed such that the compatibility of the enzyme to commonly
encountered ingredients of such compositions is increased. Alternatively, the
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43
variant may be designed such that the optimal pH, bleach or chelant stability,
catalytic activity and the like, of the enzyme variant is tailored to suit the
particular cleaning application.
In particular, attention should be focused on amino acids sensitive to
oxidation in
the case of bleach stability and on surface charges for the surfactant
compatibility. The isoelectric point of such enzymes may be modified by the
substitution of some charged amino acids, e.g. an increase in isoelectric
point
may help to improve compatibility with anionic surfactants. The stability of
the
enzymes may be further enhanced by the creation of e.g. additional salt
bridges
and enforcing metal binding sites to increase chelant stability.
Preferably, the detergent tablets of the present invention will comprise a
buffering
agent together with the pectate lyase enzyme. Such buffering agents might be
required to generate the optimum pH for activity of the pectate lyase. Any
standard buffering agent can be used. Preferred are those having optimum
buffer capacity at the pH where the pectate lyase shows optimum activity.
Examples include NaH2P04, NaHC03, Na2C03 and Citric Acid,
tris(Hydroxymethyl)aminomethane (Trizma (TM) from Sigma), triethanol amine,
NN,bis(2-Hydroxyethyl)glycine, N-tris(Hydroxymethyl)methyl-3-aminopropane-
sulfonic acid and/or mixtures thereof. Such buffering agent is typically
present at
5% or less by weight of the rapid, pectate lyase containing-dissolving phase."
Method of washinu
The compositions of the invention may be used in essentially any washing or
cleaning methods, including soaking methods, pretreatment methods and
methods with rinsing steps for which a separate rinse aid composition may be
added.
The process of the invention is conveniently carried out in the course of the
cleaning process. The method of cleaning is preferably carried out at
5°C to
95°C, especially between 10°C and 60°C. The pH of the
treatment solution is
preferably from 7 to 12.
A preferred machine dishwashing method comprises treating soiled articles
selected from crockery, glassware, silverware, metallic items, cutlery and
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44
mixtures thereof, with an aqueous liquid having dissolved or dispensed therein
an effective amount of a the herein described compositions. By an effective
amount is meant from 8g to 60g of product dissolved or dispersed in a wash
solution of volume from 3 to 10 litres, as are typical product dosages and
wash
solution volumes commonly employed in conventional machine dishwashing
methods. Preferably the detergent tablets are from 15g to 40g in weight, more
preferably from 20g to 35g in weight.
Machine laundry methods herein typically comprise treating soiled laundry with
an aqueous wash solution in a washing machine having dissolved or dispensed
therein an effective amount of the herein described compositions. By an
effective amount is meant from 20g to 300g of product dissolved or dispersed
in
a wash solution of volume from 5 to 65 litres, as are typical product dosages
and
wash solution volumes commonly employed in conventional machine laundry
methods.
Examples
The following examples are meant to exemplify compositions of the present
invention, but are not necessarily meant to limit or otherwise define the
scope of
the invention.
Dishwashing Examples
In examples I-IV, the abbreviated component identifications in the detergent
compositions have the following meanings, and all levels are quoted as parts
by
weight:
STPP : Sodium tripolyphosphate: 50% hexahydrate, 6%
Phase I and 44% Phase II
Bicarbonate : Sodium hydrogen carbonate
Citric Acid : Anhydrous Citric acid
Carbonate : Anhydrous sodium carbonate
Silicate : Amorphous Sodium Silicate (Si02:Na20 ratio =
2.0)
SKS-6 : Crystalline layered silicate of formula 8-Na2Si205
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PB1 : Anhydrous sodium perborate monohydrate
Nonionic : C13-C15 mixed ethoxylated/propoxylated fatty
alcohol with an average degree of ethoxylation
of
3.8 and an average degree of propoxylation
of 4.5,
sold under the tradename Plurafac by BASF
TAED : Tetraacetyl ethylene diamine
HEDP : Ethane 1-hydroxy-1,1-diphosphonic acid
PAAC : Pentaamine acetate cobalt (III) salt
Paraffin : Paraffin oil sold under the tradename Winog
70 by
Wintershall.
Protease : Proteolytic enzyme sold under the tradename
Savinase, Alcalase, Durazym by Novo Nordisk
A/S, Maxacal, Maxapem sold by Gist-Brocades
and proteases described in patents W091/06637
and/or W095/10591 and/or EP 251 446.
Amylase : Amylolytic enzyme sold under the tradename
Purafact Ox Am~described in WO 94/18314,
W096/05295 sold by Genencor; Termamyl~,
Fungamyl~ and Duramyl~, all available from
Novo
Nordisk A/S and those described in W095/26397
(sold under the tradename Natalase By Novo
Nordisk).
Pectate Lyase : Pectate Lyase from Bacillus agaradhaerens,
NCIMB 40482 or DSM 8721
BTA : Benzotriazole
Sulphate : Anhydrous sodium sulphate.
PEG 3000 : Polyethylene Glycol molecular weight
approximately 3000 available from Hoechst
PEG 6000 : Polyethylene Glycol molecular weight
approximately 6000 available from Hoechst
The following illustrates examples detergent tablets of the present invention
suitable for use in a dishwashing machine.
I II III IV V VI
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46
I II III IV V VI
Phase 1
STPP 9.6 9.6 10.4 9.6 9.6 11.5
Silicate 0.5 0.7 1.6 1.0 1.0 2.4
SKS-6 1.5 1.5 - 2.3 2.25
Carbonate 2.3 2.7 3.5 3.6 4.1 5.2
HEDP 0.2 0.2 0.2 0.3 0.3 0.3
PB1 2.4 2.4 2.4 3.7 3.7 3.7
PAAC 0.002 0.002 0.002 0.003 0.004 0.004
Amylase 0.1 0.1 0.11 0.2 0.2 0.2
Protease 0.06 0.06 0.06 0.09 0.09 0.09
Nonionic 0.4 0.8 0.8 1.2 1.2 1.2
PEG 6000 0.4 0.26 0.26 0.4 0.4 0.4
BTA 0.04 0.04 0.04 0.06 0.06
Paraffin 0.1 0.10 0.10 0.1 0.1 0.15
Perfume 0.02 0.02 0.02 0.01 0.01 0.01
Sulphate 0.5 0.05 2.8
Total 17.7g 18.5g 19.6g 23.Og 23.Og 23.Og
Phase 2
Pectate Lyase0.005 0.50 0.001 0.002 0.02 0.001
Amylase 0.003 0.003 0.002 0.003 0.003 0.002
Protease 0.01 0.009 0.01 0.01 0.009 0.01
Citric acid 0.3 - 0.3 0.3 - 0.30
Sulphamic - 0.3 - - 0.3 -
acid
Bicarbonate 1.1 0.4 0.4 1.1 0.4 0.4
Carbonate - 0.5 - - 0.5 -
Silicate - - 0.6 - - 0.6
CaCl2 - 0.07 - - 0.07 -
PEG 3000 0.06 0.06 0.06 0.06 0.06 0.06
Total 2.05g 2.50g 2.1 2.20g 2.02g 2.15g
g
The tablet are ared The detergent
compositions prep as active
follows.
composition phase is prepared admixingthe granular
of 1 by and liquid
components the of a
and is then die conventional
passed into rotary
press.
The
press includes itably cross-section
a punch shaped
su for
forming
a mould.
The
of the die roximately30x38
is app mm.
The
composition
is
then
subjected
to
to
a
compression kg/cm2 d the exposing
force of an punch the
940 is
then
elevated
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47
first phase of the tablet containing the mould in its upper surface. The
detergent
active composition of phase 2 is prepared in similar manner and is passed into
the die. The particulate active composition is then subjected to a compression
force of 170 kg/cm2, the punch is elevated, and the multi-phase tablet ejected
from the tablet press. The resulting tablets dissolve or disintegrate in a
washing
machine as described above within 12 minutes, phase 2 of the tablets
dissolving
within 5 minutes. The tablets display improved strength, especially on long-
term
storage, together with excellent dissolution characteristics.
Examples VII to XI
The following automatic dishwashing tablets were made in accordance with the
present invention (g of raw material and enzymes are expressed in pure enzyme)
VII VIII IX X XI
Tablet body
STPP 10.3 9.5 10.6 10.6 10.1
Carbonate 5.2 5.2 2.8 3.5 3.5
Silicate 2.4 1.6 2.9 1.6 1.1
S KS-6 2.2 2.2 - 1. 1.
5 5
HEDP 0.3 0.3 0.2 0.2 0.2
Protease 0.003 0.003 0.002 0.002 0.002
Amylase 0.001 0.001 0.001 0.001 0.001
Perborate 3.7 3.7 2.8 2.4 2.4
C13-15 EO/PO nonionic1.2 0.9 0.4 0.8 0.6
PEG4000 0.4 - - 0.3 -
PEG6000 - 0.4 - - 0.3
BTA 0.09 0.09 0.06 0.06 0.06
Paraffin 0.1 0.1 0.1 0.1 0.1
Perfume - - 0.02 0.02 0.02
Tablet body total 26.4 24.5 20.1 21.3 20.1
Dimple
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VII VIII (X X XI
Protease 0.01 0.01 0.01 0.01 0.01
Amylase 0.003 0.003 0.004 0.003 0.003
Pectate lyase 0.2 0.05 0.2 0.3 0.3
Citric 0.2 0.2 0.6 0.2 0.2
Bicarbonate 0.6 0.6 0.6 0.6 0.6
Triacetin - - 1.2 - -
PEG400 0.02 0.02 - 0.02 0.02
PEG6000 0.08 0.08 - 0.08 0.08
PEG6000 - - 1.2 - -
CaCl2 - - 0.1 - -
Dimple total 1.5 1.5 3.5 1.5 1.5
Total tablet 27.9 26.0 23.6 22.8 21.6
The following illustrates examples of detergent tablets of the present
invention
suitable for use in a laundry machine.
i) Detergent powder of compositions I-IV (see tables under) was prepared as
follows: all the particulate materials of base composition were mixed together
in a mixing drum or spray drum to form a homogenous particulate mixture.
During this mixing the spray-on of the binder system was carried out. After
this stage, the matrix was separated in two different samples. The DIBS sticky
hydrotope was added to only one of the samples and then processed
independently in a Loedige KM 600~. The layer with DIBS was used for a
harder bottom layer and the layer without DIBS was used for a softer top layer
of a dual layer tablet.
ii) Using a Bonals~ rotary press both matrices were filled in two independent
force feeding flasks. The matrix with DIBS is consecutively filled first in
the
turret stations, followed by the second matrix (the without DIBS matrix). Both
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49
layers are compressed together in the pre-compression and compression
stations to form a dual layer tablet with a harder bottom layer.
iii) In this particular example, the tablets have a rectangular cross section
of 62.5
by 38.5 mm, a height of 20.5 mm and a weight of 48 gr. The height of the
bottom layer corresponded to 25% of the total height of the tablet. If a round
tablet is made of the bottom layer matrix with the same density as in the
rectangular tablet (983 g/I), the tensile strength of the layer is 7.8 kPa.
Using
the same experiment (for a density of 991 g/I), the top layer of the tablet
has
an equivalent tensile strength of 5.1 kPa. Elasticity measurements gave
values of 1.8 J/kN for the top layer and 3.3 J/kN for the bottom layer.
Presented below are Examples for base particulate material composition for
making laundry detergent tablets according to the invention, whereby a harder
layer may be more compressed than a softer layer, or whereby different
compositions may be used or adapted for each layer.
The enzymes levels are expressed by pure enzyme by weight of the total
composition and unless otherwise specified, the detergent ingredients are
expressed by weight of the total compositions.
I II III IV
Anionic Agglomerates 1 21.0 21.0 8.6 31.5
Anionic Agglomerates 2 12.6 12.6 22.0 -
Nonionic agglomerates - - 9.1 -
Cationic Agglomerate 5.4 5.4 4.6 5.0
Layered Silicate 10.8 10.8 9.7 11.5
Sodium percarbonate 14.2 14.2 12.2 16.2
Bleach activator agglomerates5.5 5.5 6.1 4.7
Sodium carbonate 13.8 12.6 7.3 3.3
Sodium bicarbonate - - - 2.0
Sodium sulfate - - - 2.4
EDDS/Sulphate particle 0.5 0.5 0.5 0.5
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I II III IV
Tetrasodium salt of Hydroxyethane0.7 0.7 0.6 0.8
Diphosphonic acid
Soil Release Polymer 0.3 0.3 0.3 0.3
Fluorescer 0.2 0.2 0.2 0.1
Zinc Phthalocyanide sulphonate0.02 0.02 0.03 0.02
encapsulate
Soap powder 1.4 1.4 1.2 -
Suds Suppressor 1.9 1.9 2.8 2.1
Citric acid 7.1 7.1 5.5 2.0
Pectate Lyase 0.008 0.008 0.001 0.01
Protease 0.03 0.03 0.04 0.03
Lipase 0.003 0.003 0.004 0.0003
Cellulase 0.0001 0.0001 0.0001 0.0001
Amylase 0.009 0.09 0.009 0.005
Binder Spray-on-system 1 1.3 2.5 - -
Binder Spray-on-system 2 - - 3.05 -
Polymer particle - - - 3.0
Nonionic spray-on system - - - 5.2
Zeolite - - - 6.2
Perfume Spray-on - - 0.5 0.3
Perfume encapsulates - - - 0.2
Sodiumdi isoalkylbenzene sulfonate- - 2.1
TOTAL 100.00 100.00 100.00 100.00
Anionic agglomerates 1 comprise of 40% anionic surfactant, 27% zeolite and
33% carbonate.
Anionic agglomerates 2 comprise of 40% anionic surfactant, 28% zeolite and
32% carbonate.
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Nonionic agglomerate comprise 26% nonionic surfactant, 6% Lutensit K-HD 96,
40% Sodium acetate anhydrous, 20% carbonate and 8% zeolite.
Cationic agglomerates comprise of 20% cationic surfactant, 56% zeolite and 24%
sulphate.
Layered silicate comprises of 95% SKS 6 and 5% silicate.
Bleach activator agglomerates comprise of 81 % TAED, 17% acrylic/maleic
copolymer (acid form) and 2% water.
Ethylene diamine N,N-disuccinic acid sodium salt/Sulphate particle comprise of
58% of Ethylene diamine N,N-disuccinic acid sodium salt, 23% of sulphate and
19% water.
Zinc phthalocyanine sulphonate encapsulates are 10% active.
Suds suppressor comprises of 11.5% silicone oil (ex Dow Corning); 59% of
zeolite and 29.5% of water.
Binder spray-on system 1 comprises of 50% Lutensit K-HD 96 and 50% PEG
(polyethylene glycol).
Binder spray-on system 2 comprises of 0.5 parts of Lutensit K-HD 96 and 2.5
parts of PEGs.
Perfume encapsulates comprise 50% perfume and 50% starch.
Polymer particle comprises 36%, 54% zeolite and 10% water
The Nonionic spray-on system comprises of 67% C12-C15 AE5 (alcohol with an
average of 5 ethoxy groups per molecule), 24% N-methyl glucose amide and 9%
water.
Protease is selected from: proteolytic enzyme sold under the tradename
Savinase, Alcalase, Durazym by Novo Nordisk A/S, Maxacal, Maxapem sold by
Gist-Brocades and proteases described in patents W091/06637 and/or
W095/10591 and/or EP 251 446 and/or mixutres thereof.
Amylase is selected from: Amylolytic enzyme sold under the tradename Purafact
Ox AmR described in WO 94/18314, W096/05295 sold by Genencor; Termamyl
~, Fungamyl~ and Duramyl~, all available from Novo Nordisk A/S and those
described in W095/26397(Sold under the tradename Nataiase by Novo Nordisk
A/S) and/or mixtures thereof.
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52
Lipase is selected from: Lipolytic enzyme sold under the tradename Lipolase,
Lipolase Ultra by Novo Nordisk A/S and Lipomax by Gist-Brocades, and/or
mixutres thereof.
Pectate lyase from Bacillus agaradhaerens, NCIMB 40482 or DSM 8721.
Cellulase is selected from: Cellulytic enzyme sold under the tradename
Carezyme, Celluzyme and/or Endolase by Novo Nordisk A/S; and/or mixtures
thereof.
