CA2217292A1 - Detergent composition comprising a mutant amylase enzyme and oxygen bleaching agent - Google Patents

Abstract

Soaking compositions are disclosed which comprise a bleach, a builder; an anionic surfactant, a proteolytic enzyme; and a stability enhanced amylase enzyme. A process of soaking fabrics is also disclosed, wherein said fabrics are immersed in a soaking liquor comprising water and an effective amount of the composition above.

Description

W o96/31583 PCTrUS96103340 DETE RGENT COMPOSITION COMPRISING A MUTANT AMYLASE ENZYME AND
OXYGEN BLEACHING AGENT

Technical Field The present invention relates to the cleaning of fabrics in soaking conditions, i.e. in co, .dilions where the fabrics are left to soak in a soaking liquor comprising water and deterge,.l ingredients, either as a first step before a typical washing G~,~,dtion, or as a single step.
Backqround Fabric soaking o~aralions have been describad in the art. In such soaking opar~tions, fab.ics are left in co--la~;t with a soaking liquor for a prolonged 30 period of time ranging from a few hours to overnight. This laundering processhas the ad~,ant~ye that it ~--a,.i...i es the cG,-tact time between the fabrics and the key active ingredients of the soaking liquor. It also has the advantage thatit reducss or eliminates the need for a typical laundering operation involving ~ the need for mechanical agitation, or that it improves the efficiency of the 3s s~ ~hsequent typical laundering operation.

W O96/31583 PCTrUS96/03340 Such soaking operations are typically efficient to remove tough stains such as blood and food stains, which comprise a blend of proteic s~ ~hst~rlces as well as starch. Thus it is desirable that soaking compositions should comprise a protease as well as an amylase. However with such a soaking composition, 5 amylases themselves are subject to proteolytic attack, as the soaking composition is dissolved to form a soaking liquor. This is a particularly acute problem in the context of soaking, where the contact time between the ingredients in general, the amylase and the protease in particular, is generallyenormously longer than in typical laundering operation. Thus it is one object of10 the present composition to provide soaking compositions which comprise an amylase and a protease, and where the amylase will resist proteolytic attack for a significant period of time.

Another requirement of soaking cG",positions is that they should con~prise an oxygen bleach, preferably a high amount of such a bleach. Indeed, an oxygen bleach finds suitable conditions for efficacy in a soaking operalion where, contact time between the actives of the compositions and the soiled fabrics is maximized. However, the presence of bleach is also detrimental to the stability of the enzymes throughout the period of the soaking operation. Indeed 20 bleaches, especi~'ly at high level tend to denature the qualer,.a,y stucture of the enzymes, ~ osi, ~y amylase to an even more aggressive proleolytic attack.

Still a"~tl,er requirement of soaki"g co",posilions is that they should comp,isesiyllirlcalll amounts of builders. Indeed the presence of builders is necess~ry 2s to effectively control water hardness. Builders are even more necess~ry in soaking condiliG-,s where contac~ time between fabrics and wash solution is longer, as co",pared to normal washes, thus hardl,ess must be controlled for a much longer time. To comply with the peculiarities of the soaking process, soaking co.-~l~osili~"s must therefor~ conlai" high levels of builders. However,30 the pr~sence of high levels of builders is also delri",ental to the action ofenzymes. Indeed, these builders tend to bind all the calcium p,-ese"t in the soak sol ~tion, a".ongst other things, while some calcium is needed to ensure appro~,. ia~e enzyme activity.

3s Finally, anotl .er requi(e" ,enl of soaking co" ~rositions is that they should co".~rise siy,-iricant amounts of an a..ionic slJIrd~lant. A l~refer,ed anionic su,rd.,lanl class, for cost and pelrur,.,a,lce reass,.s is alkyll,e",e,-e sulpl,onale, W O 96131583 PCTrUS96/03340 particularly linear ones. Certain anionic surfactants, linear alkylbenzene sulphonates (LAS) in particular, however appear to be detrimental to enzyme stability.

5 It is thus an object of the present invention to provide a soaking compositionsuitable for a soaking opercilion, which co"".,ises an amylase in co~-,binalion with a protease, a bleaching system, a high amount of a builder, and an anionic su-rdutant preferably LAS wherein said amylase remains active for a long period of time consitent with the characteristics of a soaking operation.

SummarY of the invention In one ei,lbodi",ent the present invention enco,n~Asses compositions comprising:

- an oxy~en bleach;
- from 5% to 50% by weight of the total co,n~,osition of a builder;
- from 0.5% to 20% by weight of the total composition of an anionic su, ractanl 20 preferably linear allkyl benzene sulphonates - a proteolytic enzyme; and - a stability e,ll,a"ced amylase enzyme said stability enhancement being relative to the parenVnon-mutant form of said amylase enzyme.

25 In another embodiment the present invention encomr~-sses a process of soahi"~ rab, ics, wherein said fabrics are immersed in a soaking liquor cGIll~risill~ water and an effective amount of the cG""~osilion above/

Detailed Des.;, i,~tion of the invention ,, The presei)t invention enco""~-sses a composition and a process of soaking raL"ics. The composiliGn, hereinafter referred to as the soaking CCIIIpOSiliGII is 35 used in the soaking process.

W O 96/31583 PCTrUS96/03340 A - The comPosition:

In its first embodiment, the present invention encomp~sses a composition which comprises a bleach, a builder; an anionic surfactant, a proteolytic t enzyme; and an amylolytic enzyme.

1 - The oxY~en bleach:
As a first essential ingredient, the compositions herein comprise an oxygen bleach. Suitable oxygen bleaches in the cornposition may come from a variety of sources such as hydrogen peroxide or any of the addition compounds of hydrogen peroxide, or organic peroxyacid, or mixtures thereof. By addition 15 cc",pounds of hydloge" peroxide it is meant compounds which are formed by the ~drlition of hydrogen peroxide to a second chemical compound, which may be for example an inorganic salt, urea or organic carboxylate, to provide the ~ddition compound. Examples of the addition compounds of hydrogen peroxide include inorganic perhydrate salts, the compounds hyd~oge,) peroxide forms with or~a"ic ca,boxylates, urea, and co""~ounds in which hydrogen peroxide is clall,raled.

Examples of inorganic perhydrate salts include perborate, percarbonate, per~hospl ,ale and persilicate salts. The inorganic perhydrate salts are normally the alkali metal salts. The alkali metal salt of pe~rbo"ale, perborate or mixtures thereof, are the pre~r,ed inorganic perhydrate salts for use herein.r,ere"eJ alkali metal salt of per~,bonate is sodium perc~irLonale.

Other s~iPhle oxygen bleaches include persulphales, particularly potassium 30 persulphate K2S2O8 and sodium persulphate Na2S2O8.

Soaking compositions in the present invention may comprise from 1% to 60%
by weight of composition of a hydrogen peroxyde source, preferably from 2% to 40~~ and more ~ erer-i~ly from 15% to 40%.

~0 96/31583 PCT~US96~03340 2 - The builder As a second essential ingredient, ths compositions herein comprise a builder, or mixtures thereof, in amounts of from 5 ~/O to 50% by weight of the total co,nposition, ,~,r~rera~ly 10% to 40%, most preferably 15% to 30%. Suitable builders for use herein include inorganic as well as organic builders.

Il ,or~nic or P containing detergent builders include, but are not limited to, the alkali metal, ammonium and alkanolalr,l "o, lium salts of polyphosphates (exemplified by the tripoly~Jl,osphates, pyrophos~ les, and glassy polymeric rnetaphosphates), phosphonates, phytic acid, sili~tes, carbonates (including bicarbonates and ses~ ~rbonates), sulphates, and aluminosilicates.
However, non-phospl,dle builders are required in some locales. Illlpollallll~, 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 sili~tes, particularly those having a SiO2:Na2O ratio in the range 1:6:1 to 3:2:1 and layered silicates, such as the layered sodium silicates des~ibed in the U.S. Patent 4,664,839, issued May 12, 1987 to H.P. Rieck. NaSKS~ is the trademark for a crystalline layered silicate marketed by Hoechst (cor(""o"ly abbreviated herein as "SKS-6"). Unlike zeolite builders, the NaSKS~ silicate builder does not contain aluminum. NaSKS~ has the delta-Na2SiOs ",or~l)ology form of layered sili~ e. It can be prepared by methods such as those described in German DE-A-3,417,649 and DE-A-3,742,043. SKS~ is a highly ~.terer.ed layered si';c~te for use herein, but other such layered silic~tes, such as those having the g6neral formula NaMSixO2x 1 1 YH2O wherein M is sodium or hydrogen, x is a number from 1.9 to 4, p,efer~bly 2, and y is a number from 0 to 20, prererably0 can be used herein. Various other layered silir~tes from Hoechst include NaSKS-5, NaSKS-7 and NaSKS-11, as the alpha, beta and g~",r"a forms. As noted above, the delta-Na2SiOs (NaSKS~ form) is most prefer,ed for use herein. Other silicates may also be useful such as for example magnesium sili~te, which can serve as a crispening agent in granular formulations, as a slahil;~;ng agent for oxygen bleaches, and as a co""~G,)ent of suds control systems.

W O96/31583 PCTrUS96/03340 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 deterge,)t formulations. Aluminosilicate builders include 0 those having the empirical formula:

Mz[(zAl02)y] xH2O

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.~, and x is an integer from about 15 to about 264.

Useful aluminosilicPtes can be crystalline or amorphous in structure and can be naturally-occurring aluminosilicates or synthetically derived. A mtehod for producing aluminosilicate ion excl,a,)ge materials is ~lisclQsed in U.S. Patent 20 3,985,669, Krummel, et al, issued October 12, 1976. Prefel,ed synthetic crystalline aluminosilir~te ion exchange "~alerials useful herein are available under the designations Zeolite A, Zeolite P(B), Zeolite MAP and Zeolite X. In an esperi-'ly prefe"~d el~bo-Jiment, the crystalline aluminosilir~te ion excl ,ange material has the formula:
Na12[(Alo2)12(sio2)12l 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.
30 Pldrerably, the aluminosili~te has a particle size of about 0.1 -10 microns in dia",eter.

Organic dete,ye~)t builders suitable for the purposes of the present invention include, but are not restricted to, a wide variety of polycarboyxlate compounds.35 As used herein, pol~,ca,L,oxylate refers to co""~ounds having a plurality of ylate groups, prefer~bly at least 3 ca,Loxylates. Polyc~,L.oxylate builder can gen~rally be added to the co""~osilion in acid form, but can also be added in the forrn of a neutralized salt. When utili~ed in salt form, alkali metals, such as sodium, pot~ssium, and lithium, or alkanola"""onium salts are ~,refer,~d.

Inçluded among the polycarboxylate builders are a variety of ~te~ories of s useful materials. One i",p~,lanl r~tegory of polycarboxylate builders ~"~",I .~sses the ether pol~,cd,6Oxylates, including oxydisur~inate, as ~lislcosed in Berg, U.S. Patent 3,128,287, issued April 7, 1964, and Lamberti etal, U.S. Patent 3,635, 830, issued January 18, 1972. See also '~MS/TDS"
builders of U.S. Patent 4,663,071, issued to Bush et al, on May 5, 1987.
Suitable ether polycarboyxlates also include cyclic compounds, particularly alicyclic cGr"pounds, such as those described in U.S. Patents 3,923,679;

3,835,163; 4,120,874 and 4,102,903.

Other useful deter~ency builders include the ether hydroxypolycarboxylates, 15 such as 1, 3, 5-trihydroxy bel ,~ene-2, 4, 6-trisulphonic acid, and ca, bo~ymethyloxysuccinic acid, the various alkali metal, a" " "onium and substituted amm GI .ium salts of polyacetic acids such as ethylenediamine telr~celic acid and nitrilotriacetic acid, as well as polycarboxylates such as mellitic acid, succinic acid, oxydisu~inic acid, polymaleic acid, benzene 1,3,5-20 l. i~. L,o~ylic acid, ca, bo)cymethyloxysuccinic acid, and soluble salts thereof.

Other useful pol~,ca,L,oxylate buiders for use herein include builders accGrdinyto formula I

R1 yp ( GH2 - CR3--) R2 CO2M n wherein Y is a ~GIllollGlller or ~r"ono,.,er mixture; R1 and R2 are bleach- and alkali-stable polymer-end groups; R3 is H, OH or C14 alkyl; M is H, alkali metal, alkaline earth metal, a"",lol,ium or substituted a"",~onium; p is from 0 to 3s 2; and n is at !east 10, or mixtures thereof.

W O96/31583 PCTrUS96/03340 Prerer-ed polymers for use herein fall into two categories. The first ~tegory belongs to the class of copolymeric polymers which are formed from an unsaturated polycarboxylic acid such as maleic acid, cil,~conic acid, itaconic acid, mesAconic acid and salts thereof as first monomer, and an unsaturated 5 mG. .ocarboxylic acid such as acrylic acid or an alpha -C1 4 alkyl acrylic acid as second monomer. Referring to formula I hereinabove, the polymers belonging to said first class are those where p is not 0 and Y is selected from the acids listed hereinabove. Prerer.ed polymers of this class are those according to formula I hereinabove, where Y is maleic acid. Also, in a preferred 10 e,.,bGdi-..ent, R3 and M are H, and n is such that the polymers have a molecular weight of from 1000 to 400 000 atomic mass units.

The secG. ~d category of prefer-ed polymers for use herein belongs to the class of polymers in which, refer-i-,g to formula I hereinabove, p is 0 and R3 is H orC14 alkyl. In a ~,,efer,ed embodiment n is such that the polymers have a molecular weight of from 1000 to 400 000 atomic mass units. In a highly prerer.ed e,--bodi.nent, R3 and M are H.

The alkali-stable polymer end groups R1 and R2 in formula I hereinabove 20 suitably include alkyl groups, oxyalkyl groups and alkyl carboxylic acid groups and salts and esters thereof.

In the above, n, the degree of polymerization of the polymer can be cJeter",inedfrom the weight average polymer molecular weight by dividing the latter by the 25 average monomer molecular weight. Thus, for a maleic-acrylic copolymer having a weight average molecular weight of 15,500 and comprising 30 mole % of maleic acid derived units, n is 182 (i.e. 15,500 1 (116 x 0.3 + 72 x 0.7)).
Te,nperalure controll2d columns at 40~C agai.,sl sodium polystyrene SUIpl-lGl ~ale polymer aldl ,-Jards, available from Polymer Laboralo, ies Ud., St-r~psl,ire, UK, the polymer standards being 0.15M sodium dihycl~oge"
phos~l,ale and 0.02M tetramethyl ammonium hycl~oxide at pH 7.0 in 80120 waler/acelG"itrile.

35 Of all the above, highly prefer,ed polymers for use herein are those of the first ~e~. y in which n averages from 100 to 800, prererably from 120 to 400.

W 09613~583 P ~ flUS96~3340 Citrate builders, e.g. citric acid and soluble salts thereof (particularly sodium salt), are polyca, L,oxylate builders of particular importance due to their availability from renewable resources and their biodegradability. Citrates can also be used in combination with zeolite and/or layered silicate builders.
5 OXY~;SI ~ ,ales are also especially useful in such compositions and co" Ibinalions.

Also suitable in the deter~e"l c~l"~,osilions of the present invention are the 3,3 clica,Loxy~-oxa-1,6-hexanedioates and the related cGl"pounds disclosed in U.S. Patent 4,566,984, Bush, issued January 28, 1986. Useful succinic acid builders include the Cs-C20 alkyl and alkenyl succinic acids and salts thereof.
A particularly preferred compound of this type is dodecenylsuccinic acid.
Specific examples of succ;"ale builders include: laurylsucc;"dle, myristylsuccil ,ale, palmitylsucci"ale, 2-dodecenylsuccinate (prefe" ed), 2-~15 p~nl~decenylsucc;nale, and the like. Laurylsucc;nales are the prefer, ed builders of this group, and are described in European Patent Application 86200690.5/0,200,263, published November 5, 1986.

Other suitable polycarboxylates are disclqsed in U.S. Patent 4,144,226, ,~o 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-C18 monocarL,o,~ylic acids, can also be incorporated into the co, nposilions alone, or in combination with the aforesaid builders, 2s especially citrate and/or the succina~e builders, to provide additional builder activity.

In situ~ti~ns where pl,ospl)Gius-based builders can be used, and especially in the formulation of bars used for hand-lau"dering operaliGns, the various alkali metal ~ s~l,a~es such as the well-known sodium tripol~phosphales, sodium py. opl)ospl .ale and sodium orthopl .ospl -ate can be used. Phosphonate builders such as ethane-1-hydroxy-1, 1-diphosphonate and other known pl~ospl~G~Iates (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.

W O 96/31583 PCTrUS96/03340 3 - The anionic SU, ra.;tai ll As a third essential element, the compositions herein comprise an anionic surfactant or mixtures thereof, in amounts of from 0.5% to 20% by weight of the s total composition, preferably 3% to 15%, most prererably 5% to 10%.

Suitable anionic s~" raclanl for use herein include water soluble salts or acids of the formula ROSO3M wherein R preferably is a C1 o-C24 hydrocarbyl, ~.referal~ly an alkyl or hydroxyalkyl having a C10-C20 alkyl component, more preferably a C12-C1g alkyl-or hydroxyalkyl, and M is H or a cation, e.g., an alkali metal cation (e.g., sodium, potassium, lithium), or ammonium or substituted am",onium (e.g., methyl-, dimethyl-, and trimethyl ammonium cations and quaternary a",mo"ium cations, such as tetramethyl-ammonium and dimethyl piperdinium caliG"s and quarternary a")"~G"ium cations derived from alkylamines such as ethylamine, diethylamine, triethylamine, and mixtures thereof, and the like). Typically, alkyl chains of C12-16 are preferred for lower wash temperatures (e.g., below about 50~C) and C16-1g alkyl chains are prererldd for higher wash temperatures (e.g., above about 50~C).

Other suitable anionic sulrdctallts for use herein are water soluble salts or acids of the formula RO(A)mSO3M wherein R is an unsubstituted C10-C24 alkyl or hydroxyalkyl group having a C10-C24 alkyl component, pr~rerably a C12-C20 alkyl or hydroxyalkyl, more prererably C12-C1g alkyl or hydroxyalkyl, A is an ethoxy or propoxy unit, m is greater than zero, typically between about 2s 0.5 and about 6, more p(efera~ly between about 0.5 and about 3, and M is H or a cation which can be, for example, a metal cation (e.g., sodium, poPssi~rn, lithium, calcium",.ay"esium, etc.), ammonium or substituted-ar"mGuium cation.
Alkyl ethoxylated sulfates as well as alkyl propoxylated sulfates are conte,npldled herein. Speciflc examples of s~hstitl~te~l a~ G~ium cations include methyl-, dilllelllyl-, trimethyl-all",lonium and qualer"d,y a"~menium calions, such as t~t~a~elhyl-a~mGI)ium~ dimethyl piperdir.ium and cations derived from alkanolamines such as ethylamine, diethylamine, triethylamine, mixtures thereof, and the like. Exemplary su,raclanls are C12-C1g alkyl polyethoxylate (1.0) sulfate, C12-C1gE(1.0)M), C12-C1g alkyl polyethoxylate (2.25) sulfate, C12-C1gE(2.25)M), C12-C1g alkyl polyethoxylate (3.0) sulfate C12-C18E(3.0)~ and C12-C18 alkyl polyethoxylate (4.0) sulfate C12-C1 gE(4.0)M), wherein M is convenie, .lly selected from sodium and roPssi! Im.

CA 022l7292 l997-l0-02 fi~31S83 PCTAUS96/0334 Other anionic surfactants useful for detersive purposes can also be used herein. These can include salts (including, for example, sodium, potassium, a~l~m~ ~ium, and substituted ammonium salts such as mono-, di- and s triethanolamine salts) of soap, Cg-C2~ linear alkylbe"~e,lesulfonates, Cg-C22 primary or secondary alkanesulfonates, Cg-C24 olefinsulfonates, sulfonated polycarboxylic acids prepared by sulfonation of the pyrolyzed product of alkaline earth metal citrates, e.g., as described in British patent specification No. 1,082,179, Cg-C24 alkylpolyglycolethersulfates (containing up to 10 moles of ethylene oxide); alkyl ester sulro-,ales such as C14 16 methyl ester sulfonates; acyl glycerol sulfonates, fatty oleyl glycerol sulfates, alkyl phenol ethylene oxide ether sulfates, paraffn sulfonates, alkyl phosphates, isethionates such as the acyl isethionates, N-acyl taurates, alkyl succir,dl"ales and sulfosl~inales, monoesters of sulfosuccinate (especi-lly saturated and 115 unsaturated C12-C18 monoesters) diesters of sulfosuccinate (especially saturated and unsaturated C6-C1 4 diesters), acyl sarcosinates, sulfates of alkylpol~saccl)arides such as the sulfates of alkylpolyglucoside (the nonionic nonsulfated compounds being described below), branched primary alkyl sulfates, alkyl polyethoxy carboxylates such as those of the formula ;!0 RO(CH2CH20)kCH2COO-M+ wherein R is a C8-C22 alkyl, k is an i"leger from 0 to 10, and M is a soluble salt-f~""ing cation. Resin acids and hydrogenated resin acids are also suitable, such as rosin, hydrogenated rosin, and resin acids and hyclro~~el~ated resin acids present in or derived from tall oil. Further examples are given in "Surface Active Agents and Detergenls" (Vol. I and ll by ;~5 Schwartz, Perry and Berch). A variety of such sulrdclants are also gel)erdlly disrlQse-l in U.S. Patent 3,929,678, issued Decelnber 30, 1975 to Laughlin, et al. at Column 23, line 58 through Column 29, line 23 (herein i"cGr~,oraled by refer~, .ce).

~o Highly ~Jrere.,ed s~"racla.-ls for use in the co",positions herein are the alkyl ber~ene sulronates, espsciq~ly the linear ones.

4 - The Protease -'35 As a fourth essenlial element, the cG"~,uositions herein comprise a protease or mixtures thereof. Protease enzymes are usually prese"l in ~l~rerer, ed Lodi...e.lts of the invention at levels sufficient to provide from 0.005 to 0.2 W O96/31583 PCTrUS96/03340 Anson units (AU) of activity per gram of composition. The proteolytic enzyme can be of animal, veyetable or, prererably microorganism prefer,ed origin.
More prefer,ed is serine proteolytic enzyme of bacterial origin. Purified or nonpurified forms of enzyme may be used. Proteolytic enzymes produced by s chemically or genetically modified mutants are incl~lded by definition, as areclose structural enzyme variants. Particularly prerer,ed by way of proteolytic enzyme is bacterial serine proteolytic enzyme obtained from Bacillus, Bacillus subtilis and/or Bacillus licheniformis. Suitable commercial proteolytic enzymes include Alc~l~se ~, Esperase ~, Durazym ~, Savinase ~, M~Y~t~-se ~, M~ AI ~, and Maxapem (~ 15 (protein engineered M~Y~I); Purafect ~ and subtilisin BPN and BPN' are also commercially available. Preferred proteolytic enzymes also encompass modified bacterial serine proteases, such as those des~ibed in European Patent Application Serial Number 87303761.8, filed April 28, 1987 (particularly pages 17, 24 and 98), and which is called herein 15 r, otease B", and in European Patent Application 199,404, Venegas, published October 29, 1986, which refers to a modified bacte, ial serine proteolytic enzyme, which is called "r,otease A" herein. More prefer,ed is what is called herein "Protease C", which is a triple variant of an alkaline serine protease from Bacillus in which tyrosine replaced valine at position 104, serine20 repl ~d aspa(ag;ne at position 123, and alanine replaced threonine at pgsilioll 274. Protease C is described in EP 90915958.4, corresponding to WO 91 /06637, Published May 16, 1991, which is incorporated herein by refere,.ce. Ge--~lically modified variants, particularly of Protease C, are alsoincluded herein.
2s Also s~it~hle for use herein is a protease herein rerened to as "Protease 1~"
which is a c~lLGn~l hydrolase variant having an amino acid sequence not found in nature, which is derived from a percursor carbonyl hydrolase by s~hstitllting a dirr~r~nl amino acid for a plurality of amino acid residues at a30 pOSitiGII in said c~lL.GI~I hydrolase equivalent to posilion +76 in combination with one or more amino acid residue positions equivalent to those selected from the gorup ccnsisliny of +99, +101, +103, +107 and +123 in Bacillus amylolique~ciQns subtilisin as described in the concurrently flled patent applicaliGIls of A. Baeck, C.K. Ghosh, P.P. Greycar, R.R. Bott and L.J. Wilson, enlilled "Protease-Col.lai--i"y Cleaning Colllposilio"s" having U.S. Serial No.
08/136,797 (P&G Case 5040), and "Bleaching Compositions Comprising W O 96/31~83 PCTAUS96/~3340 Protease Enzymes" having U.S. Serial No. 08/136,626, which are incorporated herein by rerere,lce.

Some prefer~ed proteolytic enzymes are selected from the group consisting of Savinase ~, Esperase ~, M~Y~C~I ~, Purafect ~, BPN', Protease A and Protease B, and mixtures thereof. Bacterial serine protease enzymes obtained from Bacillus subtilis and/or Bacillus licheniformis are prefer,ed. Particularlyprefe, ~ed are Savinase @), Al~l~se ~, Protease A and Protease B.

5 - The amYlase As a fifth essential element, the compositions herein comprise an amylase.
Engineering of enzymes for improved stability, e.g. oxidative stability is known.
See, for example J. Biological Chem., vol. 260, No. 11, June 1985, pp 6518-6521.

"Reference amylase" hereinafter refers to an amylase outside the scope of the amylase compGne"t of this invention and against which stability of any amylase 20 within the invention can be measured.

The presenl invention thus makes use of amylases having improved stability in cJelergenls, especi-lly improved oxidative stability. A convenient ~hsolute stability refer6"ce-point against which amylases used in the instant invention 2s ,~,~,r~s~nt a measurable improvement is the stability of TERMAMYL (R) in co."",ercial use in 1993 and available from Novo Nordisk AIS. This TERMAMYL (R) amylase is a "reference amylase". Amylases within the spirit and scope of the present invention share the cl ,aracterislic of being "stability-t7nl,a"~J amylases, characteri ed, at a minimum, by a measurable 30 improvement in one or more of: oxidative stability, e.g. to hydrogen peroxide/lel~aacelylethylenediamine in buffered sol~tion at pH 9-10; thermal stability, e.g. at col"",on wash temperatures such as about 60~C; or alkaline stability, e.g. at a pH from about 8 to about 11, all measured versus the above-idel)ried re~erel)ce-amylase. Preferred amylases herein can del "onsl~ ale 35 further improvement versus more challenging reference amylases, the latter r~f~r~l ,ce amylases being illusl, aled by any of the precursor amylases of which the amylases within the invention are variants. Such precursor amylases may CA 022l7292 l997-l0-02 ll,e",selves be natural or be the product of genetic engineering. Stability can be measured using any of the art~isclosed technical tests. See references disclosed in WO 94/02597, itself and documents therein referred to being incorpated by reference.
In general, stability-enhanced amylases respecting the invention can be obtair,ed from Novo Nordisk A/S, or from Genencor International.

rrerer,ed amylases herein have the ~"~n,onality of being derived using site-directed mulagenesis from one or more of the Bacillus amylases, especially the Bacillus alpha-amylases, regardless of whether one, two or multiple amylase strains are the immediate precursors.

As noted, "oxidative slal,ilily~llha.lced amylases are prere~,ed for use herein.Such amylases are non-li",ilingly illusl,aled by the following:

(a) An amylase according to the hereinbefore incorporated WO/94/02597, Novo Nordisk A/S, published February 3, 1994, as further illustrated by a mutant in which substitution is made, using alanine or ll ,reG"ine (prererably threonine), of the methionine residue located in position 197 of the Bacjllus licheniformis alpha-amylase, known as TERMAMYL (R), or the homologous posiliGn variation of a similar parent amylase, such as Bacillus amyloliquefaciens, Bacillus subfilis, or Ri3cPlus stearothermophilus;
(b) Stability-enl ,a"ced amylases as described by Genencor Inter"dlional in a paper entitled "Oxidatively Resistant alpha-Amylases" prese"led at the 207th ~"erican Chemical Society National Meeting, March 13-17, 1994, by C. Mitchinson. Therein it was noted that bleaches in automatic dishwashing dete~ents inactivate alpha-amylases but that improved oxiddtive stability amylases have been made by Genencor from Bacillus licheniformis NCIB8061. Methionine (Met) was identified as the most likely residue to be modified. Met was substituted, one at a time, in posilions 8,15,197,256,304,366 and 438 leading to specific mutants, particularly i",pG,lant being M197L and M197T with the M197T variant being the most stable expressed variant. Stability was measured in WO 96~31~83 PCI~/US9610334 CASCADE (R) and SUNLIGHT (R); Such enzymes are commercially available from Genencor under the trade name Plurafact Oxam ~.

(c) Particularly ~referred herein are amylase variants having additional modification in the immediate parent available from Novo Nordisk AIS.
These amylases do not yet have a tradename but are those referred to by the supplier as QL37+M197T. Such enzymes are commercially available under the trade name SP 703 from Novo.

10 Any other oxidative stability-enhanced amylase can be used, for example as derived by site~irected mutagenesis from known chimeric, hybrid or simple mutant parent forms of available amylases.

6 - OPtionals:
The soaking com~,osilion of the presenl invention may further comprise a variety of other ingredients.

Optional but prefer. ed ingredients for use herein are bleach activators.
20 Examples of suitable compounds of this type are disclosed in British Patent GB
1 586 769 and GB 2 143 231. Prerer,ed examples of such compounds are tel. acet~l ethylene diamine, (TAED), sodium 3, 5, 5 trimethyl hexd.,oyloxyL.er,~ene sulphonale, diperoxy dode~noic acid as described for inslance in US 4 818 425 and nonylamide of peroxyadipic acid as described for 25 il ,alanc~ in US 4 259 201 and n-nonanoyloxybenzenesulphonate (NOBS), and acetyl triethyl citrate (ATC) such as desaibed in Ewopea,l patent application 91870207.7. Also particularly prefer,ed are N-acyl caprolactam selected from the group co"sisling of substituted or unsubstituted benzoyl caprolactam, octanyl caprola.,ld.n, nonanoyl caprolactam, hexanoyl caprolactam, decanoyl 30 caprola~a---, undecenoyl caprolactam, formyl caprolactam, acetyl caprolactam,~,ropa. Ioyl ~rola.,ld~ ", butanoyl caprolactam pentanoyl caprolactam. The soaking co~"~osilions herein may comprise mixtures of said bleach activators.
.~
r, efe" ed mixtures of bleach activators herein CG m ,ul ise n-. IGl .a. Ioyloxybel ,~el ,esulpl ,Gndle (NOBS) together with a second bleach activator having a low tendency to generate diacyl peroxide, but which delivers mainly perac;d. Said second bleach activators may include tet,dcelyl ethylene W O96/31583 PCTrUS96/03340 diami,)e (TAED), acetyl triethyl citrate (ATC), acetyl caprolactam, benzoyl caprolactam and the like, or mixtures thereof. Said mixtures of bleach activators are preferably used in the embodiment of the present invention where the soaking liquors are controlled to a pH below 9.5. Indeed, it has been found that mixtures of bleach activators comprising n-nonanoyloxybenzenesulphonate and said second bleach activators, allow to boost particulate soil cleaning performance while exhibiting at the same time good pe.r~r"~a"ce on diacyl peroxide sensitive soil (e.g. beta carotene) and on peracid sensitive soil (e.g. body soils).

Accordingly, the soaking compositions herein may comprise from 0% to 15%
by weight of the total composition of n-nonanoyloxybenzenesulphonate, preferably from 1% to 10% and more preferably from 3% to 7% and from 0% to 15% by weight of the total co"~position of said second bleach activator prefe(ably from 1% to 10% and more preferably from 3% to 7%.

It may be desirable for them to further comprise chelating agents which help to control the level of free heavy metal ions in the soaking liquors, thus avoidingrapid decG" ,position of the oxygen released by said source of available 20 oxygen. Suitable amino carLoxylate chelating agents which may be used herein include diethylene triamino pentacetic acid, ethylenediamine tel, ~Ac~t~les (EDTA), N-hydroxyethylethylenediamine tri~cet~tes, nitrilol, i~c~t~es, ethylenediamine telrapru~rionales, triethylenetet, aamine lIeX~AC t~es, and ethanoldiglycines, alkali metal ammonium and substituted 25 am,..Gn.lJm salts ll ,er~of or mixtures thereof. Further suitable chelating agents include ethylenedia"line-N,N'~ s~inic acids (EDDS) or alkali metal, alkaline earth metal, ammonium, or substituted ammol.ium salts thereof. Particularly suitable EDDS co",pounds are the free acid form and the sodium or ~syllesium salt or complex thereof. Also others s~it~hle chelating agents may be the Of ~anic pl ,ospl .ol ,ales, including amino alkylene poly(alkylene pl ,os,cl .onale), alkali metal ethane 1 -hydroxy diphospl)onales, nitrilo l,i",elh~lene phospl-onales, ethylene diamine tetra methylene pl,ospl,ona~es and diethylene triamine penta methylene ~I-os~hGnates. The phospl,Gnale ~,."~ounds may be present either in their acid form or in the form of their metal 35 alkali salt. r~ererably the organic pl,ospl,o.)ate compounds where present are in the form of their magnesium salt.

~NO96131583 PCT~US96/03340 The soaking compositions in the present inventlon may accordingly co",,u,ise from 0% to 5% by weight of the total compositions of said chelating agents, ,c,referdbly from 0% to 3%, more ~referably from 0.05% to 2%.

s Soaking compositions in the present invention may further comprise other Gptional ingredients such as additional surfactants, other than anionics, fillers, optical L,ri~l,le,lers, additional enzymes, soil release agents, photoactivated bleaches such as Zn phthalocyanine sulphonale, dyes, dye transfer inhibitors, pigments and perfumes. Said optio"al ingredients can be added in varying 10 amounts as desired.

The compositions herein can be manufactured in solid, preferably granular, or liquid form.

B - The Process:

In a second embodiment, the present invention encomp~sses a process of soaking fabrics. As used herein, the expression ",urocess of soaking faL,rics"
~o refers to the action of leaving fabrics to soak in a soaking liquor col"~,ising water and a co" ,1 ~osilion as described hereinabove, for a period of time sufficient to clean said fabrics. The soaking process can be performed i..dep~r,denlly from any other process, such as a typical laundering operation, or a first step before a second, typical lau"deri"g step. In the prefe"ed ;2s soaki"y processes of the invention, fabrics are left to soak for a period of time ranging from 10 minutes to 24 hours, prefera61y from 30 min to 24 hours, most p(efe(aL,ly 4 hours to 24 hours. After the fabrics have been i"""er~ed in said soaking liquor for a sufficient period of time, they can be removed and rinsed with water. The fablics can also be washed in a normal lau"dering operation after they have been soaked, with or without having been rinsed inbetween the soaking operatiG,. and the sl ~hsequent launderir,g operalion.

In the soaking process herein, a soaking cGI"posilion des~ibed hereinabove is diluted in an appropriate amount of water to produce a soaking liquor. Suitable doses may range from 45 to 50 grams of soaking cor"posilion in 3.5 to 5 liters of water, down to 90 to 100 grams of soaking co""~osition in 20 to 45 liters of water. Typically one dose is 45-50 grams in 3.5 to 5 Lt for a conce, Itl aled soak W O96131583 PCT~US96103340 (buckeVsink). For washing machine soaks the dose is 90-100 grams in about 20 (Europe) to 45 (US) liter of water. The fabrics to be soaked are then i"""er~ed in the soaking liquor for an apl~r~priate period of time.

There are factors which may influence overall performance of the process on partic~JI~'e dirt/soils. Such factors include prolonged soaking time. Indeed thelonger fabrics are soaked the better the end results. Ideally soaking time is ove",i~l,t i.e. 12 hours up to 24 hours. Another factor is the initial warm or warmluke tei "peralure. Indeed higher initial temperatures of the soaking 10 liquors ensure large benefits in performance.

ExamPles The following compositions are made by mmixing the listed ingredients in the listed pro~o,lions. Soaking liquors are formed by diluting each time 45 9 of said compositions in between 3.5 lit. to 5.0 lit. of water. 0.5 to 2 Kg of fabrics are then eadt time immersed in said soaking liquor for a time ranging from 10 minutes to 24 hours. Finaliy the fabrics are removed from the soaking liquors rinsed with water and washed with a regular washing process handwash or 20 washing machine wash with a regular detergent with or without re-using the soaki"~ liquor then said fabrics are left to dry.

Inqredient 1 2 3 (% wlw) (% wlw) (% wlw) Plurafact Oxam ~ or SP 703 0.29 0.29 0.29 (~4~~ active protein) Protease B; ~4% active protein) 0.26 0.52 0.52 Sodium p6,borate ."onol.ydrate 29 29 29 Sodium percarbonale 0 0 O
Citric acid 11 11 11 7eolit~A 12 12 12 Polyacrylate (Acusol 445ND) 10 10 10 Sodium silicate (~,.,or~,hous; 1.6r) 0.3 0.3 0.3 Anionic (LAS) 7 7 7 TAED ~ ~ 5 DTPA 0.2 0.2 0.2 Other inerts and minors up to up to up to W 096131583 PCT~US96/03340 Inqredient 4 5 6 (% wlw) (% wlw) (% wlw) Plurafact Oxam ~ or SP 703 0.26 0.26 0.26 (~4% active protein) Protease (B; ~4% active protein) 1.00 0.50 0.50 Sodium perborate monohydrate 0 0 0 Sodium perca(LGIld~e 31 31 31 Citric acid 10 10 10 Zeolite A 0 0 0 Polyacrylate (Acusol 445ND) 11 11 11 Sodium silicate (ar"or~hous; 1.6r) 0.4 0.4 0.4 Anionic(LAS) 4 4 4 DTPA 0.2 0.2 0.2 Other, inerts and minors up to upto upto

Claims (12)

Claims

1 A composition comprising:

- an oxygen bleach;
- from 5% to 50% by weight of the total composition of a builder;
- from 0.5% to 20% by weight of the total composition of an anionic surfactant;
- a proteolytic enzyme; and - a stability enhanced amylase enzyme, said stability enhancement being relative to the parent/non-mutant form of said amylase enzyme.

2. A composition according to claim 1 wherein said anionic surfactant is a linear alkyl benzene sulphonate.

3. A composition according to the any of the preceding claims which comprises 3% to 15% of said anionic surfactant, preferably 5% to 10%.

4. A cleaning composition according to the preceding claims wherein said stability-enhanced amylase enzyme is oxidative stability-enhanced.

5. A cleaning composition, according to claim 4 wherein said oxidative stability is enhanced from substitution using threonine of the methionine residue located in position 197 of Bacillus licheniformis or the homologous position variation of a similar parent amylase.

6. A composition according to the any of the preceding claims which comprises 10% to 40% of said builder, preferably 15% to 30%.

7. A composition according to any of the preceding claims which comprises from 1% to 60% of said oxygen bleach, preferably from 15% to 40%.

8. A composition according to any of the preceding claims wherein said oxygen bleach is an alkali metal salt of perborate or percarbonate.

9. A composition according to any of the preceding claims which further comprises a bleach activator.

10. A composition according to any of the preceding claims wherein said protease is Savinase ~, Alcalase ~, Protease A and Protease B.

11. A process of soaking fabrics, wherein said fabrics are immersed in a soaking liquor comprising an effective amount of a composition according to any of the preceding claims, for an effective period of time, then removed from said soaking liquor.

12. A process according to claim 11 wherein said time ranges from 10 minutes to 24 hours.