MXPA99002827A - Liquid detergents containing proteolytic enzyme and protease inhibitors - Google Patents

Abstract

Aqueous liquid detergent compositions are described which comprise a proteolytic enzyme wherein the proteolytic activity is reversibly inhibited by a peptide protease inhibitor selected from the group consisting of aldhehydes and trifluoromethyl ketones.

Description

LIQUID DETERGENTS CONTAINING PROTEOLYTIC ENZYME AND PROTEASE INHIBITORS TECHNICAL FIELD The present invention relates to liquid detergent compositions containing enzymes. Very specifically, the present invention relates to liquid detergent compositions containing a detersive surfactant, a proteolytic enzyme, and a protease peptide inhibitor selected from the group consisting of peptide aldehydes and peptide trifluromethyl ketones.

BACKGROUND OF THE INVENTION Aqueous liquid detergents containing protease are well known, especially in the context of laundry. A problem commonly found in such liquid aqueous detergents that contain protease is the phenomenon of degradation by the proteolytic enzyme of second enzymes in the composition, such as amylase, lipase, and cellulase, or in the same protease. As a result, the stability of the second enzyme or protease in the detergent composition is affected and the detergent composition consequently yields less.

In response to this problem, several protease inhibitors or stabilizers have been proposed. For example, several protease inhibitors or stabilizers have been proposed. For example, several references have proposed the use of the following compounds to aid in the stabilization of enzymes. Benzamidine hydrochloride, lower aliphatic alcohols or carboxylic acids, mixtures of a polyol and a boron compound, esters of aromatic borate and calcium, particularly calcium formate. It has recently been discovered that certain peptide aldehydes and peptide trifluromethylketone act to stabilize the protease enzyme. Although such compounds have been used to vary the success in liquid detergents, they are not without problems. For example, certain peptide aldehydes can be expensive and create complexities for formulators, especially for liquid detergents. Other inhibitors such as calcium and boric acids are more economical but do not stabilize the enzymes as well as the peptide aldehydes. It is therefore an object of the present invention to provide alternating peptide aldehydes and inhibitors of trifluoromethyl ketone protease which are effective and suitable for use in liquid detergent compositions. TECHNICAL BACKGROUND The use of several protease inhibitors or stabilizers has been proposed. For example, the document E.U.A. 4,566,985 proposes the use of benzamidine hydrochloride; EP 376 705 proposes the use of lower aliphatic alcohols or carboxylic acids; EP 381, 262 proposes the use of a mixture of a polyol and a boron compound; and EP 91870072.5 proposes the use of aromatic borate esters. See also U.S. Patent No. 5,030,378 issued July 9, 1991. See also documents E.U.A. 4,261, 868; E.U.A. 4,404,115; E.U.A 4,318,818; and EP 130,756. The use of peptide derivatives for protein inhibition seems to have been described in therapeutic applications. EP 293,881 discloses the use of peptide boronic acids as inhibitors of serine proteases similar to tripcin. Documents EP 185,390 and E.U.A. 4,399,065 describe the use of certain peptide aldehyde derivatives for the inhibition of blood coagulation. J 90029670 describes the use of optically active alpha-aminoaldehydes for the inhibition of enzymes in general. See also "Inhibition of Trombin and Trypsin by Tripeptide Aldehydes", Int. J. Peptide Protein Res., Vol. 12 (1978), pp. 217-221; Gall, Bacsy & Rappay, and "Tripeptide Aldehyde Protease Inhibors May Depress in Vitro Prolactim and Growth Hormone Relase" Endocrinology, Vol. 116, No. 4 (1985), pp. 1426-1432; Rappay, Makara, Bajusz & Nagy. Certain peptide aldehydes have been described in EP-A-473 502 to inhibit skin irritation mediated by protease. See in particular EP 185,390 WO94 / 04651, published on March 3, 1994 WO94 / 04652, published March 3, 1994, EP 583,536, published February 23, 1994, EP 583,535, published on February 3, 1994. 1994, EP 583,534, published on February 23, 1994, WO93 / 13125, published July 8, 1993, US 4,528,525, USA 4,537,706, E.U.A. 4,537,707 and E.U.A. 5,527,487.

BRIEF DESCRIPTION OF THE INVENTION The invention herein is the liquid detergent composition comprising: a) an effective amount of a detersive surfactant; b) an active proteolytic enzyme; and c) a protease peptide inhibitor having the formula: Z-B-NH-CH (R) -C (O) -X wherein B is a peptide chain comprising 2 to 5 amino acid portions; X is hydrogen or CF3; Z is an N-blocking portion selected from the group consisting of sulfonamides, phosphoramidates, thioureas, sulfonamides, sulfonic acids, phosphinamides, thiocarbamates, amidophosphates, sulfamoyl derivatives and phosphamides; R is selected from the group consisting of straight branched C1-C6 unsubstituted alkyl, phenyl, and C7-C9 alkylaryl moieties. The preferred compositions further comprise a source of calcium ion or boric acid.

Preferably, the liquid detergent compositions herein comprise, by weight of the composition: a) from about 1 to about 95%, preferably from about 8% to about 70% of said detersive surfactant; b) from about 0.0001% to about 5%, preferably from about 0.003% to about 0.1%, of an active proteolytic enzyme. c) from about 0.0001% to about 5%, preferably from about 0.0001 to about 1%, most preferably from about 0.0006% to about 0.5% of the described peptide protease inhibitor; d) optionally, from about 0.01% to about 1%, preferably about 0.05% to about 0.5% calcium ion; and e) optionally, from about 0.25% to about 10%, preferably from about 0.5% to about 5% boric acid or a compound capable of forming boric acid, preferably diol. The proteolytic enzyme useful herein is preferably a protease of the subtilisin type and can be selected from the group consisting of alcalase R, subtilisin BPN ', protease A, protease B and mixtures thereof.

The calcium ion source for use herein is preferably selected from the form of calcium, calcium xylene sulfonate, calcium chloride, calcium acetate, calcium sulfate and mixtures thereof. The dishwashing compositions herein may also contain adjunct detersives, including but not limited to one or more of the following: foam impellers, chelators, polyacrylate polymers, dispersing agents, colorants, perfumes, processing aids, and mixtures thereof. In addition to dishwashing compositions, the liquid detergent compositions may further comprise an effective amount of amylase enzyme. Additionally, dishwashing compositions may optionally comprise an effective amount of a boric acid source and a diol. Typically, the dishwashing compositions optionally, preferably, will comprise from about 0.25% to about 10%, preferably from about 0.5% to about 5%, most preferably from about 0.75% to about 3% by weight of boric acid or a compound capable of forming boric acid and a diol, i.e., 1,2-propondiol. In a preferred embodiment for heavy duty detergent compositions useful in laundry washing, the liquid detergent composition further comprises an effective amount of one or more of the following enzymes: lipase, amylase, cellulase and mixtures thereof. Preferably for laundry compositions, the second enzyme is lipase and is obtained by cloning Humicola Lanuainosa and expressing the gene in Aspergillus Orvzae. The lipase is used in an amount of about 10 to about 18,000 units of lipase per gram, preferably from about 60 to about 6,000 units per gram. In another preferred composition useful for washing clothes, the second enzyme, is a cellulase derivative of Humicola Lanuainosa and is used in an amount of about 0.0001% to about 0.1% by weight of the total composition of said cellulase. The compositions herein may also contain adjunct detersives, including but not limited to one or more of the following: foam impellers, builders, soil release polymers, polyacrylate polymers, dye transfer agents, dye transfer inhibitors, dyes, perfumes, processing aids, brighteners and mixtures thereof. Additionally, for laundry washing compositions, the detersive surfactant is typically present in an amount of about 10% to about 70% by weight of the total composition. In addition, the laundry compositions may optionally comprise an effective amount of a boric acid source and a diol. Typically laundry compositions optionally, but preferably comprise from about 0.25% to about 10%, preferably from about 0.5% to about 5%, most preferably from about 0.75% to about 3% by weight of boric acid or a compound capable of forming boric acid and a diol, i.e., 1,2-propanediol. All per hundred and proportions herein are by weight, and all references cited are incorporated herein by reference, unless specifically indicated otherwise.

DETAILED DESCRIPTION OF THE INVENTION Definitions. The present detergent compositions comprise an "effective amount" of a "spot removal enhancement amount" of individual components defined herein. An "effective amount" or "amount of stain removal enhancement" is capable in any amount to measurably improve dirt removal or removal of stains from a substrate, i.e., dirty cloth or dirty dishes, when washed by the consumer. In general, said amount may vary widely. The liquid aqueous detergent compositions according to the present invention comprise three essential ingredients: (A) a protease peptide inhibitor selected from the group consisting of aldehydes and trifluoromethyl ketones, a mixture thereof, as described herein, (B) a proteolytic enzyme or a mixture thereof; and (C) a detersive surfactant. The compositions according to the present invention preferably further comprise (D) a source of calcium ions (E) a second enzyme compatible with the detergent or a mixture thereof, (F) boric acid and a diol, and furthermore can understand (G) other optional ingredients. Protease Peptide Inhibitors.- The detergent compositions according to the present invention comprise as a first essential ingredient, a protease peptide inhibitor selected from the group consisting of aldehydes and trifluoromethyl ketones, or mixtures thereof, having the formula: ZB-NH- CH (R) -C (O) -X wherein B is a peptide chain comprising 2 to 5 amino acid portions; X is hydrogen or CF3; Z is an N-blocking portion selected from the group consisting of phosphoramidate [(R "O) 2 (O) P-], sulfenamide [(SR") 2-], sulfonamide [(R "(O) 2S-], sulfonic acid [SO3H], phosphinamide [(R ".2 (0) P-], sulfamoyl derivative [R" O (O) 2S-], thiourea [(R ") 2N (0) C-], thiocarbamate [ R "O (S) C-], phosphonate [R" -P (O) OH], and amidophosphate [R "O (OH) (O) P-], wherein each R" is independently selected from the group consisting unsubstituted straight or branched C ^ -CQ alkyl, phenyl, C7-C9 alkylaryl, and cycloalkyl portions, wherein the cycloalkyl ring may be C4-C8 and may contain one or more heteroatoms selected from the group consisting of , N and S, and R is selected from the group consisting of unsubstituted straight or branched C- | -C6 alkyl, phenyl and C7-C0 alkylaryl moieties.

Preferred portions R are selected from the group consisting of methyl, isopropyl, secbutyl, isobutyl, -C6H5, -CH2-C6H5, and -CH2CH2-C5H5, which respectively can be derived from the amino acids Ala, Val, He, Leu, PGIy (phenylglycine), Phe and HPhe (homophenylalanine) by converting the carboxylic acid group to an aldehyde or trifluoromethyl ketone group. Although such portions are not amino acids (and may or may not have been synthesized from an amino acid precursor), for purposes of simplifying the exemplification of inhibitors useful herein, the aldehyde portion of the inhibitors is indicated as derived from amino acids by the addition of "H" after the analogous amino acid [ie, "-AlaH" represents the chemical moiety "-NHCH (CH3) C (O) H"]. Trifluoromethyl ketones are similarly represented by the addition of "CF3" after the analogous amino acid (ie, "-AlaCF3" represents the chemical moiety "-NHCH (CH3) C (O) CF3".] Preferred peptide chains B are they are selected from the group consisting of peptide chains having the amino acid sequences according to the general formula: Z-A5-A-A3-A2-A1-NH-CH (R) -C (O) -X Thus, the following amino acids, when present, are: A1 is selected from Ala, Gly; A2 is selected from Val, Ala, Gly, He; A3, if present, is selected from Phe, Leu, Val, He; A4 if present, is any amino acid, but is preferably selected from Gly, Ala; A5 if present, is any amino acid, but preferably Gly, Ala, Lys. The aldehydes of the present invention can be prepared from the corresponding amino acid wherein the C-terminal end of said amino acid is converted from a carboxylic group to an aldehyde group. Said aldehydes can be prepared by known methods, for example as described in documents E.U.A. 5015627, EP 185930, EP 583,534, and DE 3200812. The trifluoromethyl ketones of the present invention can be prepared from the corresponding amino acid wherein the C-terminal end of said amino acid is converted from a carboxylic group to the trifluoromethyl ketone group. Said trifluoromethyl ketones can be prepared by known methods, for example, as described in EP 583,535. Although not intended to be limited by theory, it is believed that the protease peptide inhibitors according to the present invention bind to the proteolytic enzyme in the liquid detergent composition, thereby inhibiting said proteolytic enzyme. With dilution in water, the proteolytic activity is restored by the dissociation of the proteolytic enzyme inhibitor peptide protease complex.

The N-terminal end of said protease inhibitors according to the present invention are protected by one of the protecting groups of the N-blocking portion selected from the group consisting of sulfonamides, phosphonamides, thioureas, sulfenamides, sulfonic acids, phosphinemides, thiocarbamates, amidophosphates and phosphonamides. However, in a highly preferred embodiment of the present invention, the N-terminus of said protease inhibitor is protected by a methyl, ethyl or benzylsulfonamide [CH3SO2-; CH3CH2SO2-; or C6H5CH2SO2-], and methyl, ethyl or benzyl amidophosphate [CH3? (OH) (O) P-; CH 3 CH 2 O (OH) (O) P-; or groups C6H5CH2O (OH) (O) P-]. The synthesis of N-blocking groups can be found in the following reference: Protective Groups in Oraanic Chemistrv. Greene, T., Wuts, P., John Wiley & Sons, New York, 1991, pp 309-405; March, J, Advanced Organic Chemistrv. Wiley Interscience, 1985, pp. 445, 469, Carey, F. Sundberg, R., Advanced Organic Chemistry, Part B, Plenum Press, New York, 1990, p. 686-89; Atherton, E., Sheppard, R., Solid Phase Peptide Svnthesis, Pierce Chemical, 1989,? P.3-4; Grant, G., Svnthetic Peptides. W.H. Freeman & Co. 1992, pp. 77-103; Stewart, J., Young, J., Solid Phase Peptide Svnthesis. 2nd Edition, IRL Press, 1984, pp. 3,5,11,14-18, 28-29. Bodansky, M., Principies of Peptide Svnthesis. Springer-Verlag, 1988, pp. 62,203, 59-69; Bodansky, M., Peptide Chemistrv. Springer-Verlag, 1988, pp. 74-81, Bodansky, M., Bodansky, A., The Practice of Peptide Svnthesis. Springer-Verlag, 1984, pp. 9-32.

Examples of protease inhibitors for use herein are: CH3S02Phe-Gly-Ala-Leu-H, CH3S02Val-Ala-Leu-H, C6H5CH2? (OH) (O) P-Val-Ala-Leu-H, C6H5CH2O (OH) (O) P-Val-Ala-Leu-CF3, CH3CH2S? 2-Phe-Gly-Ala-Leu-H, C6H5CH2SO2 -Val-Ala-Leu-H, C6H5CH2O (OH) (O) P-Leu-Ala-Leu-H, C6H5CH2O (OH) (O) P-Phe-Ala-Leu-H, CH3? (OH) (O) P-Leu-Gly-Ala- Leu-H. In the synthesis examples of the subsequent methods are described to synthesize certain methods of said peptide protease inhibitors.

EXAMPLE OF SYNTHESIS 1 Synthesis of tripeptide trifluoromethyl ketone Moc-Phe-GIv-Ala-LeuCFi (a) N-trityl-leucine methyl ester: To a solution of 2.50 g (13.8 mmol) of Leu-OMe.HCl in 100 ml of CH2Cl2 was added 3.86 ml of TEA (27.5 mmol) by dripping. After the addition was complete, 3.76 g (13.5 mmol) of triphenylmethyl chloride in 15 ml of CH2Cl2 was added dropwise. The mixture was stirred for 4 h. The solution was diluted with 5% EtOAc / petroleum ether and washed with water. The organic phase was dried (MgSO 4), filtered and the solvent removed. The residue was chromatographed on silica to give 4.8 g of pure product (90% yield). (b) N-trityl-leucine: To a cold (0 ° C) solution of 4.70 g (12.2 mmol) of N-trityl-leucine methyl ester in 100 ml of THF was added 28.1 ml of a 1.5 M solution of diisobutylaluminum hydride (42.2 moles) in THF by dripping. The solution was stirred for 6 h at this temperature and the reaction was quenched with saturated Na-K tartrate, extracted with EtOAc, dried (MgSO 4), filtered and the solvent removed. 4.13 g of the desired material that was used without purification was recovered. To a solution of 1.29 g (14.9 mmol) of oxalyl chloride in 20 ml of CH2Cl2 at -78 ° C was added 2.26 ml of DMSO (29.8 mmol) in 5 ml of CH2Cl2 by dropping. After the addition was complete, 4.13 g (1 1.5 mmol) of crude N-trityl-leucinol in 10 ml of CH2Cl2 was added. The solution was warmed to 0 ° C and poured into a mixture of water and ether. The phases were separated and the ether phase was dried (MgSO4) and evaporated to obtain 1.37 g of the desired compound. (c) 5-Methyl-3-tritylamino-1,1,1-trifluoro-2-hexanol: to a solution of 1.37 g (3.83 mmol) of N-trityl-leucinal and 0.653 ml (4.59 mmol) of CF3TMS in THF 0.121 g (0.383 mmol) of tetrabutylammonium fluoride trihydrate was added in one portion. The solution was stirred for 3 h at room temperature and stirred into the solvent. The residue was dissolved in EtOAc, washed with water, dried (MgSO 4), and the solvent was removed to obtain 1.20 g of the product which was subjected to silica chromatography (0.760 g of pure product). (d) 3- (N- (Cbz-Gly-Ala)) - 5-methyl-1,1,1-trifluoro-2-hexanol: to a solution of 1.21 g (2.83 mmoles) of 5-methyl-3- tritylamino-1,1,1-trifluoro-2-hexanol in 10 ml of dioxane was added 5 ml of 4.0 M HCl in dioxane. The solution was stirred for 2 h at room temperature and the solvent was removed.

The residue was triturated with ether and filtered on solid material. The resulting HCl salt (0.627 g, 2.83 mmol) was suspended in 10 mL of CH2Cl2 and Z-Gly-Ala-OH (0.793 g, 2.83 mmol) was added. To this mixture was added 0.870 ml (6.23 mmoles) of TEA followed immediately by the addition of 0.473 ml (3.12 mmoles) of DEPC. The mixture was stirred overnight and the solvent was removed. The residue was dissolved in EtOAc, and washed with 1N HCl, saturated NaHCO 3, and brine. The product solution was dried (MgSO 4), filtered and the solvent removed to give 1.06 g of the product. (e) 3- (N- (Ms-Phe-Gly-Ala)) - 5-mehyl-1,1,1-trifluoro-2-hexanol: To a solution of 1.06 g (2.37 mmoles) of 3- (N -Cbz-Gly-Ala) -5-methyl-1, 1,1-trifluoro-2-hexanol in 5 ml of MeOH 0.35 g of Pd / C were added. The mixture was gassed and hydrogenated under positive hydrogen pressure overnight. The mixture was filtered with Celite and the solvent was removed. The residue was dissolved in CH2Cl2 and 0.898 g (22.37 mmoles) of added Ms-Phe-OH. To this mixture was added 0.732 ml (5.22 mmoles) of TEA, followed by the addition of 0.395 ml (2.61 mmoles) of DEPC. The solution was stirred overnight and the solvent was removed. It was subjected to silica chromatography to achieve 0.720 g of pure product. (f) Ms-Phe-Gly-Ala: to a mixture of 1.59 g (3.75 mmoles) of Dess-Martin pyriodinano in 15 ml of CH2CI2 were added 0.650 g (1.25 mmoles) of 3- (N-Ms-Phe-Gly) -Ala) -5-methyl-1,1,1-trifluoro-2-hexanol in 5 ml of CH 2 Cl 2 and the mixture was stirred for 3 h. To this mixture was added 6.51 g (2.2 mmoles) of Na 2 S 2+ 3 in saturated NaHC 3 3 and the resulting solution was stirred for 10 minutes. The solution was extracted with EtOAc and the organic phase was dried (MgSO 4), filtered and the solvent was removed. The residue was chromatographed on silica to obtain 0.445 g of pure product.

EXAMPLE OF SYNTHESIS 2 Synthesis of Ms-Phe-GIv-Ala-LeuH (a) Ms-Phe-Gly-OH: to a solution of 2.0 g (9.0 mmoles) of Phe-Gly-OH, which was dissolved in 9 ml of 1N NaOH and cooled to 0 ° C, 0.766 were added simultaneously ml (9.9 mmol) of methanesulfonyl chloride and 9 ml of 1N NaOH, in separate addition tunnels. After the addition was complete, the reaction was stirred for 15 minutes at 0 ° C and 1 h at room temperature. At this point, the solution was cooled to 0 ° C, the pH was adjusted to 9.5 and washed with EtOAc (1X, 50 ml). The aqueous phase (0 ° C) was then adjusted to pH = 2.5 (2N HCl) and extracted with EtOAc (3X, 50 ml), dried (MgSO 4), filtered, and solvent removed to achieve 2.0 g of the pure product. (b) Ms-PheGly-Ala-Leucinol: a solution was prepared by dissolving 0.500 g (1.67 mmoles) of N-Ms-Phe-Gly-OH in 15 ml of THF, cooling it to -50 ° C and adding 0.366 ml ( 3.33 mmoles) of NMM followed by 0.216 ml (1.67 mmoles) of isobutylchloroformate. Said solution was stirred for 5 minutes and 0.374 g (1.67 mmoles) of Ala-Leucinol.HCl was added in a mixture of 10 ml of THF and minimal DMF. Stirring was continued at 0 ° C for 15 minutes and 2 h at room temperature. The solution was quenched with 5 ml of 1N HCl, extracted with EtOAc (3X, 50 ml), the combined extracts were washed with saturated NaHCO 3 and saturated NaCl. The resulting organic phase was then dried (MgSO 4), filtered, evaporated and chromatographed on silica to yield 0.260 g of the desired material. (c) Ms-Phe-Gly-Ala-LeuH: a solution was prepared by adding 0.337 g (0.798 mmoles) of Dess-Martin periodinone to 5 ml of CH2CI2 and stirring for 10 minutes. To this solution 0.125 g (0.266 mmoles) N-Ms-Phe-Gly-Ala-Leucionol was added in one portion. The reaction was continued until the TLC showed complete conversion at which time the solution was poured into 25 ml of saturated NaHCO 3 containing 1.8 g (5,586 mmoles) Na 2 S 2 3- 3. After stirring for 10 minutes, the mixture was extracted with EtOAc (3X, 50 ml). The combined extracts were dried (MgSO 4), evaporated, and subjected to silica chromatography to obtain 0.048 g of the product.

Gly = glycine Ala = alanine Leu = leucine Phe = phenylalanine Orne = methyl ester TEA = triethylamine DECP = diethylcyanophosphonate TLC = thin layer chromatography MeOH = methanol Pd / C = palladium on activated carbon EtOH = ethanol THF = tetrahydrofuran Ms = methanesulfonyl Proteolytic Enzyme.- Another essential ingredient in current liquid detergent compositions is the proteolytic enzyme. Also included are proteolytic enzyme mixtures. The proteolytic enzyme may be of animal, vegetable or microorganism origin (preferred). Proteases for use in the detergent compositions herein include (but are not limited to) trypsin, subtilisin, chymotrypsin and elastase-type proteases. Preferred for use herein are proteolytic enzymes of the subtilisin type. Particularly preferred is the bacterial serine proteolytic enzyme obtained from Bacillus subtilis and / or Bacillus licheniformis. Protease enzymes are usually present in said liquid detergent compositions at levels sufficient to provide 0.005 to 0.1 Anson units (AU) of activity per gram of composition. Suitable proteolytic enzymes include Novo Industri A / S Alcalase® (preferred), Esperase®, Savinase® (Copenhagen, Denmark), Gist-brocades' Maxatase®, Maxacal® and Maxapem 15® (protein manufactured by Maxacal®) (Delft, Netherlands), and subtilisin BPN and BPN '(preferred), which are commercially available. Preferred proteolytic enzymes are also modified bacterial serine proteases, such as those made by Genencor International, Inc. (San Francisco, California), which are described in European Patent 251, 446, issued April 28, 1987 (particularly pages 17, 24 and 98), and which are referred to herein as "Protease B", and the US Patent 5,030,378, Venegas, issued July 9, 1991, which relates to a modified bacterial serine proteolytic enzyme (Genencor International) which is called "Protease A" herein (like LBP '). See in particular columns 2 and 3 of the U.S. Patent. 5,030,378 for a complete description, including the amino sequence, of protease A and its variants. The preferred proteolytic enzymes, therefore, are selected from the group consisting of Alcalase® (Novo Industri A / S), BPN ', Protease A and Protease B (Genencor), and mixtures thereof. Protease B is the most preferred. Another preferred protease, referred to as "Protease D" is a variant of carbonylhydrolase having an amino acid sequence that is not found in nature, which is derived from a carbonilase precursor by replacing a different amino acid with a plurality of amino acid residues. at a position in said carbonylhydroxysa equivalent to the +76 position, preferably also together 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 Bacillus amyloliquefaciens subtilisin numbering, as described in WO 95/10615 published April 20, 1995 by Genencor International. Useful proteases are also described in PCT publications: WO 95/30010 published November 9, 1995 by The Procter & Gamble Company; WO 95/30011 published November 9, 1995 by The Procter & Gamble Company; WO 95/29979 published November 9, 1995 by The Procter & Gamble Company.

Detersive Surfactant.- Another essential ingredient of the present invention is a detersive surfactant in an effective amount, typically from about 1 to 95, preferably from about 8 to 70% by weight. The detersive surfactant can be selected from the group consisting of anionic, nonionic, cationic, ampholytic, zwitterionic surfactants, and mixtures thereof. The detergent compositions of the present invention can be formulated by selecting the type and amount of detersive surfactant, together with other auxiliary ingredients as described herein., to be used in the laundry context. Therefore, the particular surfactants used can vary widely. The benefits of the present invention are especially notable in compositions containing ingredients that are unsuitable for enzymes, such as certain detergency builders and surfactants. These include (but are not limited to) anionic surfactants such as alkylether sulfate, linear alkylbenzene sulfonate, alkyl sulfate, etc. Suitable surfactants are described below.

Anionic Surfactants A type of anionic surfactant that can be used includes alkyl ether sulfonates. These are convenient because they can be made with renewable non-petroleum resources. The preparation of the alkyl ether sulfonate surfactant component can be carried out in accordance with known methods which are described in the technical literature. For example, linear esters of C8-C20 carboxylic acids can be sulfonated with gaseous SO3 according to 'The Journal of the American Oil Chemists Society,' 52 (1975), pp. 323-329. natural fatty substances such as those derived from tallow, palm and coconut oils, etc. The preferred alkyl ether sulphonate surfactant, especially for laundry applications, comprises alkyl ether sulphonate surfactants of the structural formula: R3-CH (SO3M) -C (O) -OR4 wherein R3 is a C8-C20 hydrocarbyl > preferably an alkyl, or combination thereof, R4 is a hydrocarbyl of C-i-Cg, preferably an alkyl or combination thereof, and M is a soluble salt-forming cation. Suitable salts include metal salts such as sodium, potassium and lithium salts, and substituted or unsubstituted ammonium salts, such as methyl-, dimethyl-, trimethylammonium, and quaternary ammonium cations, e.g., tetramethylammonium and dimethylpiperidinium, and cations derived from alkanolamines, e.g., monoethanolamine, diethanolamine and triethanolamine. Preferably, R3 is C10-C16 alkyl and R4 is methyl, ethyl or isopropyl. Especially preferred are methyl estersulfonates wherein R3 is C- | 4-Ci6 alkyl- Alkyl sulfate surfactants are another type of anionic surfactants of importance for use herein. In addition to providing excellent overall cleaning capacity when used in combination with polyhydroxy fatty acid amides (see below), including good grease / oil cleaning at a wide range of temperatures, wash concentrations and wash times; the dissolution of alkyl sulphates can be obtained, as well as an improved formulability in liquid detergent formulations are water soluble salts or acids of the formula ROSO3M, wherein R is preferably a hydrocarbyl of C < |? -C24, preferably alkyl or hydroxyalkyl having a C10-C20 alkyl component. most preferably a C 12 -C 18 alkyl or hydroxyalkyl, and M is H or cation, e.g., an alkali metal cation (e.g., sodium, potassium, lithium), substituted ammonium cations such as methyl cations -, dimethyl-, and trimethylammonium and quaternary ammonium, e.g., tetramethylammonium and dimethylpiperidinium, and cations derived from alkanolamines such as ethanolamine, diethanolamine, triethanolamine and mixtures thereof, and the like. Typically, C12-C16 alkyl chains are preferred for washing temperatures for lower wash temperatures (ie, less than about 50 ° C) and C- | 6-18 alkyl chains are preferred for washing temperatures. higher (ie, greater than approximately 50 ° C). Alkoxylated alkylsulphate surfactants are another category of useful anionic surfactants. These surfactants are water soluble salts or acids typically of the formula RO (A) mS? 3M, wherein R is an unsubstituted C10-C24 alkyl or hydroxyalkyl group having a C10-C24 alkyl component. preferably an alkyl or hydroxyalkyl of C12-2O 'most preferably alkyl or hydroxyalkyl of C12-C8. A is an ethoxy or propoxy unit, m is greater than zero, typically about 0.5 and about 6, most preferably about 0.5 and about 3, and m is H or a cation which may be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.), ammonium cation or substituted ammonium. The ethoxylated alkyl sulphates and also the propoxylated alkyl sulphates are contemplated herein. Specific examples of the substituted ammonium cations include methyl-, dimethyl-, trimethylammonium and quaternary ammonium, such as tetramethylammonium, dimethylpiperidinium and cations derived from alkanolamines, e.g., monoethanolamine, diethanolamine and triethanolamine, and mixtures thereof. Exemplary surfactants are C12-I8 polyethoxylated sulfate alkyl sulfate (1.0), polyethoxylated C-12-I8 alkyl sulfate (2.25), polyethoxylated c12_c18 alkyl sulfate (3.0), and polyethoxylated C12-C18 alkyl sulfate (4.0), wherein M conveniently it is selected from sodium and potassium.

Other Anionic Surfactants Other anionic surfactants useful for detersive purposes may also be included in the laundry detergent compositions of the present invention. These may include salts (including, for example, sodium, potassium, ammonium and substituted ammonium salts, such as mono-, di and triethanolamine salts) of soap, linear C9-C20 alkylbenzenesulfonates. C8-C22 primary or secondary alcansulfonates. C8-C24 olefin sulfonates, sulfonated polycarboxylic acids prepared by sulfonation of the pyrolyzed product of alkaline earth metal citrates v.gr, as described in British Patent Specification No. 1,082,179, alkyl glycerol sulfonates, fatty acyl glycerol sulphonates, fatty oleyl glycerol sulfates, alkyl phenole sulfates of ethylene oxide, paraffinsulfonates, alkyl phosphates, isethionates such as acyl isethionates, N-acyltaurates, fatty acid amides of methyl tauride, alkylsuccinamates and sulfosuccinates, monoesters of sulfosuccinates (especially saturated and unsaturated C12-C18 monoesters) and diesters of sulfosuccinates (especially diesters) C6-C14 saturated and unsaturated), N-acyl sarcosinates, alkylpolysaccharide sulfates such as alkylpolyglucoside sulfates (the non-sulphonated nonionic compounds are described below), branched primary alkyl sulphates and alkylpolyethoxycarboxylates such as those of the Formula RO (CH2CH2?) KCH2COO'M + wherein R is a C8-C22 alkyl. k is an integer from 0 to 10, and M is a soluble salt forming cation. Resin acids and hydrogenated resin acids such as colophonic acids, hydrogenated rosin acids and resin acids and hydrogenated resin acids present in or derived from wood oil are also suitable. Additional oils are described in "Suface Active Agents and Detergents" (Vol. I and II by Schwartz, Perry and Berch). A variety of such surfactants are also generally described in the U.S. Patent. 3,929,678, issued December 30, 1975 Laughiin and others in Column 23, line 58 to Column 29, line 13 (incorporated herein by reference).

Non-ionic detergent surfactants - Suitable detergent nonionic surfactants are generally described in US Pat. 3,929,678 to Laughiin et al., Issued December 30, 1975, column 13, line 14, to column 16 line 6, incorporated herein by reference. Below are non-limiting exemplary classes of non-ionic surfactants. The condensates of polyethylene oxide, polypropylene and polybutylene of alkylphenols. These compounds include the condensation products of alkylphenols having an alkyl group containing from about 6 to about 12 carbon atoms, either in a straight chain or branched chain configuration with the alkylene oxide. In a preferred embodiment, the ethylene oxide is present in an amount of about 5 to about 25 moles of ethylene oxide per mole of alkylphenol. Commercially available nonionic surfactants of this type include Igepal ™ CO-630, marketed by GAF Corporation; and Triton ™ X-45, X-114, X-100 and X-102, all sold by Rohm & Haas Company. These compounds are commonly known as alkylphenol alkoxylates (ie, alkylphenol ethoxylates). The condensation products of aliphatic alcohols with about 1 to about 25 moles of ethylene oxide. The alkyl chain of the aliphatic alcohol may be either straight or branched, primary or secondary, and generally contains from about 8 to about 22 carbon atoms. Preferred are the condensation products of alcohols having an alkyl group containing from about 10 to about 20 carbon atoms, with from about 2 to about 18 moles of ethylene oxide per mole of alcohol. Examples of commercially available nonionic surfactants of this type include Tergitol® 15-S-9 (the linear alcohol condensation product of C-11-C15 with 9 moles of ethylene oxide), Tergitol ™ 24-L-6 NMW ( condensation product of C12-C14 primary alcohol with 6 moles of ethylene oxide with a limited molecular weight distribution), both marketed by Union Carbide Corporation; Neodo. ™ 45-9 (the linear alcohol condensation product of C14-C-15 with 9 moles of ethylene oxide), Neodol ™ 23-3 (the linear alcohol condensation product of C-12-C13 with 3.0 moles of ethylene oxide), Neodol ™ 45-5 (the linear condensation product of C14-C-15 with 7 moles of ethylene oxide), Neodol ™ 45-5 (the C14 linear alcohol condensation product -C15 with 5 moles of ethylene oxide), marketed by Shell Chemical Company, Kyro ™ EOB (the condensation product of C13-C15 alcohol with 9 moles of ethylene oxide), marketed by The Procter & Gamble Company. This category of nonionic surfactant is generally referred to as "alkyl ethoxylates". The condensation products of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The hydrophobic portion of these compounds will preferably have a molecular weight of from about 1500 to about 1800, and will exhibit insolubility in water. The addition of polyoxethylene portions to this hydrophobic portion tends to increase the water solubility of the molecule in general, and the liquid character of the product is retained to the point where the polyoxethylene content is about 50% of the total weight of the product. condensation product, which corresponds to condensation with up to about 40 moles of ethylene oxide. Examples of compounds of this type include some Pluronic ™ surfactants commercially available from BASF. Semi-polar nonionic surfactants are a special category of nonionic surfactants including water-soluble amine oxides containing an alkyl portion of about 1 to about 18 carbon atoms and 2 portions selected from the group consisting of alkyl groups and groups hydroxyalkyl containing from about 1 to about 3 carbon atoms; water-soluble phosphine oxides containing an alkyl portion of about 10 to about 18 carbon atoms and 2 portions selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; and water soluble sulfoxides containing an alkyl portion of about 10 to about 18 carbon atoms and a portion selected from the group consisting of alkyl and hydroxyalkyl portions of about 1 to about 3 carbon atoms. The semipolar nonionic surfactants include the amine oxide surfactants having the formula: R3 (OR4)? N (O) (R5) 2 wherein R3 is an alkyl, hydroxyalkyl or alkylphenyl group, or mixtures thereof, containing from about 8 to about 22 carbon atoms; R 4 is an alkylene or hydroxyalkylene group containing from about 2 to about 3 carbon atoms, or mixtures thereof; x is from 0 to about 3; and each R§ is an alkyl or hydroxyalkyl group containing from about 1 to about 3 carbon atoms, or a polyethylene oxide group containing from about 1 to about 3 ethylene oxide groups. The R5 groups may be linked to each other, eg, through an oxygen or nitrogen atom to form a ring structure. These amine oxide surfactants include in particular C <alkyldimethylamine oxides. RJ-8 and C8-C- 2- alkoxyethyldihydroxyethylamine oxides The alkylpolysaccharides described in the U.S.A. Do not. 4,565,647, Filling, issued January 21, 1986, having a hydrophobic group containing from about 6 to about 30 carbon atoms, preferably from about 10 to about 16 carbon atoms, and a polysaccharide, i.e., a polyglycoside , a hydrophilic group containing from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7 units of saccharide. Any reducing saccharide containing 5 or 6 carbon atoms can be used, i.e., the glucose, galactose and galactosyl moieties can replace the glucosyl moieties (optionally the hydrophobic moiety is attached in the 2-, 3-, 4- positions). , etc., thus giving a glucose or galactose as opposed to a glucoside or galactoside). The intersaccharide linkages can be, i.e., between position one of the additional saccharide units and positions 2-, 3-, 4- and / or 6-of the preceding saccharide units. Optionally, and less conveniently, there can be a polyalkylene oxide chain linking the hydrophobic portion and the polysaccharide portion. The preferred alkylene oxide is ethylene oxide. Typical hydrophobic groups include alkyl groups, saturated or unsaturated, branched or unbranched, with about 8 to about 18 carbon atoms, preferably about 10 to about 16 carbon atoms. The alkyl group may contain up to about 3 hydroxyl groups and / or the polyalkylene oxide chain may contain up to about 10, preferably less than 5, portions of alkylene oxide. Suitable aiquilpolysaccharides are octyl-, nonyl-, decyl-, undecyldecyl-, tridecyl, tetradecyl-, pentadecyl-, hexadecyl-, heptadecyl-, and octadecyl- di-, tri-, tetra-, and pentaglucosides, and tallowalkyltetra- , penta-, and hexa-glucosides. Preferred alkyl polyglycosides have the formula R2? (CnH2nO) t (glycosyl) x wherein R2 is selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof, wherein the alkyl groups contain from about 10 to about 18, preferably from about 12 to about 14, carbon atoms; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is from about 1.3 to about 10, preferably about 1.3 to about 3, most preferably from about 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare these compounds, the alcohol or alkylpolyethoxylated alcohol is first formed, and then reacted with glucose or a glucose source to form the glucoside (linkage at position 1). The additional glycosyl units can then be linked between their position 1 and the preceding glycosyl units in the 2-, 3-, 4- and / or 6- position, preferably predominantly in the 2-position. Fatty acid amide surfactants They have the formula: wherein Rβ is an alkyl group containing about 7 to about 21 (preferably about 9 to about 17) carbon atoms, and each R7 is selected from the group consisting of hydrogen, C1-C4 alkyl, hydroxyalkyl ? -C4, y- (C2H4) XH, wherein x ranges from about 1 to about 3. Preferred amides are ammonium amides, monoethanolamides, diethanolamides and C8-C20 isopropanolamides- Cationic surfactants.- Cationic surfactants may also be included in the detergent compositions of the present invention. Cationic surfactants include ammonium surfactants such as alkyldimethylammonium halides and surfactants having the formula: [R2 (OR3) and] [R4 (OR) and] 2R5N + X- wherein R2 is an alkyl or alkylbenzyl group having from about 8 to about 18 carbon atoms in the alkyl chain, each R3 is selected from the group which consists of -CH2CH2-, -CH2CH (CH3) -, -CH2CH (CH2OH) -, -CH2CH2CH2-, and mixtures thereof; each R 4 is selected from the group consisting of C 1 -C 4 alkyl, C 1 -C 4 hydroxyalkyl, benzyl ring structures formed by joining the two groups R 4, -CH 2 CHOH-, -CHOHCOR 6 CHOHCH 2 OH, wherein R 6 is any hexose or polymer of hexose having a molecular weight less than about 1000, and hydrogen when and not 0; R5 is the same as R4 or is an alkyl chain in which the total number of carbon atoms of R2 plus R5 is not greater than about 18; each y is from 0 to approximately 10 and the sum of the values of y is from 0 to approximately 15; and X is any compatible anion.

Other cationic surfactants useful herein are also disclosed in the U.S.A. 4,228,044, Cambre, issued October 14, 1980, incorporated herein by reference.

Other surfactants Ampholytic surfactants can be incorporated into the detergent compositions of the present invention. These surfactants can be broadly described as aliphatic derivatives of secondary or tertiary amines, or as aliphatic derivatives of heterocyclic secondary or tertiary amines in which the aliphatic radical can be a straight or branched chain. One of the aliphatic substituents contains at least about 8 carbon atoms, typically from about 8 to about 18 carbon atoms, and at least one contains an anionic water solubilization group, e.g., carboxy, sulfate, sulfonate . See the patent of E.U.A. No. 3,929,678 to Laughiin et al., Issued December 30, 1975, column 19, lines 18-35, for examples of ampholytic surfactants. Zwitterionic surfactants may also be incorporated in the detergent compositions of the present invention. These surfactants can be broadly described as derivatives of secondary and tertiary amines, derivatives of heterocyclic secondary and tertiary amines or derivatives of quaternary ammonium, quaternary phosphonium or tertiary sulfonium compounds. See the patent of E.U.A. No. 3,929,678 to Laughiin et al., Issued December 30, 1975, in column 19 line 38, to column 22 line 48, for examples of zwitterionic surfactants. In general, ampholytic and zwitterionic surfactants are used in combination with one or more anionic and / or nonionic surfactants.

Polyhydroxy fatty acid amide surfactant The liquid detergent compositions of the present invention may also contain an enzyme enhancing amount of polyhydroxy fatty acid amide surfactant. By "enzyme enhancer" is meant that the formulator of the composition can select an amount of polyhydroxy fatty acid amide to be incorporated into the compositions, which will improve the cleansing action of the enzyme in the detergent composition. In general, for conventional levels of enzymes, the incoforation of approximately 1% by weight of polyhydroxy fatty acid amide will improve the yield of the enzyme. The detergent compositions of the present invention will typically comprise about 1% by weight of polyhydroxy fatty acid amide surfactant, preferably about 3% to about 30% of the polyhydroxy fatty acid amide. The surfactant component of polyhydroxy fatty acid amide comprises compounds having the structural formula: R2-C (0) -N (R1) -Z wherein: R1 is H, C1-C4 hydrocarbyl, 2-hydroxyethyl, 2- hydroxypropyl, or a mixture thereof, preferably C 1 -C 4 alkyl, most preferably C 1 alkyl; | or C2, more preferably C- alkyl; (ie, methyl); and R2 is a C5-C31 hydrocarbyl, preferably straight-chain C5-C19 alkyl or alkenyl, most preferably straight-chain C9-C17 alkyl or alkenyl, more preferably C-l-C-1 alkyl or alkenyl < Straight chain or a mixture thereof, and Z is a polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with at least 3 hydroxyls directly connected to the chain, or an alkoxylated derivative (preferably ethoxylated or propoxylated) thereof. Z will preferably be derived from a reducing sugar in a reductive amination reaction; most preferably Z is a glycityl. Suitable reducing sugars include glucose, fructose, maltose, lactose, galactose, mannose and xylose. High-dextrose corn syrup, high-fructose corn syrup and high-maltose corn syrup, as well as the individual sugars mentioned above, can be used as starting materials. These corn syrups can produce a mixture of sugar components for Z. It should be understood that this does not mean to exclude other suitable starting materials. Z is more preferably selected from the group consisting of -CH2 (CHOH) nCH2? H, -CH (CH2OH) (CHOH) n-iCH2OH, -CH2 (CHOH) 2- (CHOR,) (CHOH) CH-2OH, and alkoxylated derivatives thereof, wherein n is an integer from 3 to 5, inclusive, and R 'is hydrogen or a cyclic or aliphatic monosaccharide. Preferred glycityls are those wherein n is 4, particularly -CH 2 (CHOH) 4CH 2 OH. R1 can be for example N-methyl, N-ethyl, N-propyl, N-isopropyl, N-butyl, N-2-hydroxyethyl, or N-2-hydroxypropyl. R2-CO-N < it can be, for example, cocoamide, stearamide, oleamide, lauramide, myristamide, capricamide, palmitamide, seboamide, etc. Z can be 1-deoxyglucityl, 2-deoxyfructityl, 1-deoxymaltityl, 1-deoxylactityl, 1-deoxygalactityl, 1-deoxyanityl, 1-deoxyltotriotityl, etc. Methods for making polyhydroxy fatty acid amides are known in the art. In general, they can be made by reacting an alkylamine with a reducing sugar in a reductive amination reaction to form a corresponding N-alkyl polyhydroxyamine, and then reacting the N-alkylpolydroxyamine with a fatty aliphatic ester or triglyceride in a condensation / amidation step to form the N-alkylamine product of N-polyhydroxy fatty acid. Methods for making compositions containing polyhydroxy fatty acid amides are described, for example, in Great Britain Patent Specification 809,060, published February 18, 1959, U.S. Pat. 2,965,576, issued December 20, 1960 to E. R. Wilson and patent of E.U.A. 2,703,798; Anthony M. Schwartz, issued March 8, 1955, and patent of E.U.A. 1,985,424, issued on December 25, 1934 to Piggott, each of which is incorporated herein by reference.

Second enzyme.- Preferred compositions herein further include an amount of yield increase of a second enzyme compatible with a detergent. By "compatible detergents" is meant the compatibility with other ingredients of a liquid detergent composition, such as a detersive tenure agent and a detergency pH enhancer. These second enzymes are preferably selected from the group consisting of delipase, amylase, cellulase and mixtures thereof. The term "second enzyme" excludes the proteolytic enzymes described above, whereby each composition having a second enzyme contains at least two kinds of enzyme, including at least one proteolytic enzyme. The amount of second enzyme used in the composition varies according to the type of enzyme. In general, these second enzymes are preferably used from about 0.0001 to 0.3, more preferably from 0.001 to 0.1% by weight. Mixtures of the same class of enzymes (for example lipases) or two or more classes (for example, cellulase and lipase) can be used. Purified or non-purified forms of enzyme can be used. Any suitable lipolytic enzyme for use in a liquid detergent composition can be used in these compositions. Suitable lipase enzymes for use herein include those of bacterial and fungal origin. Suitable bacterial lipases include those produced by microorganisms of the Pseudomonas groups, such as Pseudomonas stutzeri ATCC 19.154. as described in British Patent 1,372,034, incorporated herein by reference. Suitable lipases include those which show a positive immunological cross-reaction with the lipase antibody produced by the microorganism Pseudomonas fluorescens IAM 1057. This lipase and a method for its purification have been described in Japanese Patent Application, Application number 53-20487 of February 24, 1978. This lipase is available from Amano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name lipase P, "Amano", henceforth it will be called "Amano-P". Said lipases must show a positive immunological cross-reaction with the Amano-P antibody, using the standard and well-known immunodiffusion procedure according to Ouchterloni (Acta. Med. Sea., 133, pages 76-79 (1950)). These lipases and a method for their immunological cross-reaction with Amano-P are also described in U.S. Patent Nos. 4,707,291, Thom et al., Issued November 17, 1987, incorporated herein by reference. Typical examples thereof are Amano-P lipase, lipase ex Pseudomonas fragi FERM P 1339 (available under the trade name Amano-B), lipase Pseudomonas nitroreducens var. Lipoliticum FERM P 1338 (available under the trade name Amano-CES), lipases ex Chromobacter viscosum, and other lipases Chromobacter viscosum var. Lipoliticum NRRLB 3673. from Toyo Jozo Co., Tagata, Japan, and lipases ex Chromobacter viscosum. Suitable fungal lipases include those produced by Humicola lanuoinosa and Thermomyces lanuginosus. The most preferred lipase is obtained by cloning the Humicola lanuginosa gene and expressing the gene in Aspergillus orizae as described in the European patent application 258 068 (Novo Industri A / S), commercially available from Novo Nordisk A / S under the name commercial Lipolase®. These compositions may be used from about 10 to 18,000, preferably from about 60 to 60,000, lipase units per gram (LU / g) of lipase. One unit of lipase is that amount of lipase that produces 1 mmol of titratable fatty acid per minute in a pHstatus, where the pH is 9.0, and the temperature is 30 ° C, substrate is an emulsion of 3.3% by weight of oil of olive and 3.3% of gum arabic in the presence of 13 mmol / 1 Ca ++ and 20 mmol / 1 NaCl in 5 mmol / 1 Tris-pH enhancer. Any cellulase suitable for use in a liquid detergent composition can be used in these compositions. Suitable cellulase enzymes for use herein include those of bacterial and fungal origin. Preferably, they will have an optimum pH of between 5 and 9.5.

About 0.0001 to 0.1% by weight of cellulase can be used. Suitable cells are described in United States patent 4. 435,307, Barbesgaard et al. Issued March 6, 1984, incorporated herein by reference, which describes the fungal cellulase produced Humicola insolens. Suitable cellulases are also described in GB-A-2,075,028, GB-A-2,095,275 and DE-OS-2,247,832. Examples of said cellulases are cellulases produced by a strain of Humicola insolens (Humicola grísea var. Thermoidea). particularly the strain Humicola DSM 1800 v cellulases produced by a Bacillus fungus No cellulase 212- that produces fungus belonging to the genus Aeromonas. and cellulase extracted from the hepatopanrhias of a marine mollusk (Dolabella Auricle Solander). Any suitable amylase for use in a liquid detergent composition can be used in those compositions. For example, amylases include amylases obtained from a special strain of B. licheniformis. described in more detail in British Patent Specification No. 1. 296,839 (Novo). Aminololytic proteins include, for example Rapidase ^, International Bio-Synthesis, Inc. and Termamil ^ Novo Industries. From about 0.0001% to 0.55%, preferably 0.0005 to 0.1% by weight of amylase can be used. Calcium.- The compositions herein can optionally comprise any calcium salt soluble in water can be used as a source a source of calcium ion. Any water soluble calcium can be used, including calcium acetate, calcium formate, calcium xylene sulfonate and calcium propionate. Divalent ions, such as zinc and magnesium ions, can replace the calcium ion completely or in part. Thus for the liquid detergent compositions herein, the source of calcium ions can be replaced partially or completely with a source of another divalent ion. The useful calcium in the present is accessible by enzyme. Therefore, the claimed compositions are substantially free of sequestrants, for example polyacids capable of forming calcium complexes that are soluble in the composition. However, minor amounts of sequestrants such as polyacids or mixtures of polyacids can be used. Accessible calcium per enzyme is defined as the amount of calcium ions effectively available to the enzyme component. From a practical point of view the calcium accessible by enzyme is therefore the soluble calcium in the composition in the absence of any storage sequestrant, for example, which has a constant balance of complexation with calcium equal to or greater than 1.5 to 20 degrees C. Boric acid.- The present compositions contain from 0.25% to 10%, preferably from 0.5% to 5%, more preferably from 0.75% to 3%, by weight of boric acid or a compound capable of forming boric acid in the composition (calculated based on boric acid). Boric acid is preferred, although other compounds such as boric oxide, borax and other alkali metal borates (eg ortho-sodium, meta-, pyroborate, and sodium pentaborate) are suitable. Substituted boric acids (for example phenylboronic acid, butan boronic acid, and p-bromo phenylboronic acid) can also be used in place of boric acid. The compositions of the present invention also contain polyols, especially diols, which contain only carbon, hydrogen and oxygen atoms. These preferably contain from 2 to 6 hydroxy groups. Examples include propylene glycol (especially 1,2-propanediol, which is preferred), ethylene glycol, glierol, sobitol, mannitol, glucose, and mixtures thereof. The polyol generally represents from 1% to 15%, preferably from 1.5% to 10%, more preferably from 2% to 7%, by weight of the composition. Optional ingredients- Builders can optionally be included in the compositions here, especially for laundry compositions. Organic builders can also be used. When present, the compositions will typically include at least about 1% detergency builder and may be an inorganic or organic builder. Liquid laundry formulations preferably include from about 3% to 30%, more preferably to about 5 to 20% by weight of detergency builder. Inorganic builders include, but are not limited to, alkali metal, ammonium and alkanolammonium polyphosphate salts (illustrated by crystalline polymeric tripolisphosphates, pyrophosphates and metaphosphates), phosphonates, phytic acid, silicates, carbonates (including bicarbonates and cesicarbonates) , sulfates and aluminosilicates. Borate builders, as well as builders that contain borate-forming materials and can produce borate under detergent storage or washing conditions (hereinafter referred to as "borate builders") can also be used. "). Preferably, the non-borate builder enhancers are used in the inventive compositions that are intended to be used under washing conditions at less than about 50 ° C, especially at less than 40 ° C. Examples of silicate builders are alkali metal silicates, particularly those having an Si? 2: Na2? Ratio. in the 1.6.1 to 3.2.1 scale 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, incorporated here by reference.

However other silicates may also be useful such as for example magnesium silicate, which can serve as a brittle agent in granular formulations, as a stabilizing agent for oxygen bleaches and a component of foam control systems. Examples of carbonate builders are alkaline earth metal and alkali metal carbonates, including sodium carbonate and sesquicarbonate and mixtures thereof with ultrafine calcium carbonate as described in German Patent Application No. 2,321,001 published on November 15, 1973, the description of which is incorporated herein by reference. The aluminusilicate detergency builders are useful in the present invention. Aluminusilicate builder detergents are of great importance in most heavy duty granular detergent compositions currently marketed and could also be a significant detergent builder ingredient in liquid detergent formulations. The aluminosilicate builders include those having the empirical formula Mz (zAl? 2-ySi? 2) wherein M is sodium, potassium, ammonium or substituted ammonium, z is from about 0.5 to about 2 e and is 1; This material having a magnesium ion exchange capacity of at least about 50 mg. equivalent to hardness of CaC 3 per gram of anhydrous aluminosilicate. The preferred aluminosilicates are zeolite detergency builders having the formula: Naz [(Ai? 2) z (Si? 2) y]. XH2O Where z and y are integers of 6, the molar ratio of za Y lies on the scale of 1.0 at about 0.5 and x is an integer of about 15 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. Patents 3,985,669, Krummel, et al., Issued October 12, 1976, hereby incorporated by reference. The preferred synthetic crystalline aluminosilicate ion exchange materials useful herein are available under the designations Zeolite A, Zeolite P (B), and Zeolite X. In an especially preferred embodiment, the crystalline aluminosilicate ion exchange material has the formula Na - | 2 [(AI02) 12 (S¡O2) 12] xH2O where x is about 20 about 30, especially about 27. This material is referred to as Zeolite A.

Preferably, the aluminosilicate has a particle size of about 0.1-10 microns in diameter. Specific examples of polyphosphates are the alkali metal tripolyphosphates, sodium pyrophosphate, potassium and ammonium, sodium and potassium pyrophosphate and ammonium, sodium and potassium orthophosphate, sodium polymetaphosphate, in which the degree of polymerization has a scale of 6 to about 21 and salts of phytic acid. Examples of phosphonate builder-enhancing salts are the water-soluble salts of ethan 1-h idroxy-1,1-diphosphonate, particularly sodium and potassium salts, the water-soluble salts of methylene diphosphonic acid, for example trisodium and tripotassium salts. and the water-soluble salts of substituted methylene diphosphonic acids, such as ethylidene, isopropylidene, benzylmethyldene, and trisodium and tripotassium halomethylidene phosphonates. The phosphonate detergency builder salts of the aforementioned types are described in U.S. Patent Nos. 3,159,581 and 3,213,030, issued December 1, 1964 and October 19, 1965, by Diehl; States patent United States No. 3,422,021 issued on January 14, 1969 to Roy and patent of United States Nos. 3,400,148 and 3,422,137 issued September 3, 1968 and January 14, 1969, of Quimbi, said descriptions are incorporated herein by reference. Preferred organic detergency builders for the purposes of the present invention include a wide variety of polycarboxylate compounds. As used here, "polycarboxylate" refers to compounds having a plurality of carboxylate groups, preferably at least 3 carboxylates. The polycarboxylate builder can generally be added to the composition in acid form, but can also be added in the form of a neutralized salt. When used in salt form, the alkali metal salts of sodium, potassium and lithium or alkanol ammonium are preferred. A variety of useful materials are included among the polycarboxylate builder builders. An important category of polycarboxylate builders encompasses ether polycarboxylates. A number of ether polycarboxylates have been described for use as builders. Examples of useful ether polycarboxylate include oxydisuccinate as described in Berg, U.S. 3,128,287, issued April 7, 1964, and Lamberti et al., U.S. Patent 3,635,830, issued January 18, 1972, which are incorporated herein by reference.

A specific type of ether polycarboxylates useful as builders in the present invention also include those having the general formula: CH (A) (COOX) -CH (COOX) -O-CH (COOX) -CH (COOX (B where A is H or H, B is H or -O-CH (COOX) -CH2 (COOX), and X is H or a cation that forms a salt, for example, if in general formula above A and B were both H, then the compound would be oxydisuccinic acid and its water soluble salts.If A is OH and B is H, then the compound is monosuccinic tartrate acid (TMS) and its water soluble salts.If A is H and B is - O-CH (COOX) -CH2 (COOX), then the compound is disuccinic tartrate acid (TDS) and its water soluble salts Mixtures of these detergency builders are especially preferred for use herein.

Particularly preferred are mixtures of TMS and TDS in a weight ratio of TMS to TDS from about 97: 3 to about 20:80. These detergency builders are described in U.S. Pat. No. 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. Patent 3,923,679; 3,835,163; 4,158,635; 4,120,874 and 4,102,903, which are incorporated herein by reference. Other useful builders include the ether hydropolycarboxylates represented by the structure: HO- [C (R) (COOM) -C (R) (COOM) -O] n-H where M is hydrogen or a cation where the resulting salt is soluble in water, preferably an alkali metal, ammonium or substituted ammonium cation, n is from about 2 to about 15 (preferably n is from about 2 to about 10, more preferably n has an average of about 2 about 4) and each R is the same or different and is selected from hydrogen, C-j_4 alkyl or substituted alkyl from C-1.4 (preferably R is hydrogen), Still other ether polycarboxylates include copolymers of maleic anhydride with ethylene or methylvinyl ether, 1, 3,5-trihydroxy benzene 2,4,6-trisulfonic acid and carboxymethyloxysuccinic acid. Organic polycarboxylate builders also include various alkali metal, ammonium and substituted ammonium salts of polyacetic acids. Examples include the sodium, potassium, lithium, ammonium and substituted ammonium salts of ethylenediamine tetraacetic acid and nitrilotriacetic acid. Also included are polycarboxylates such as melific acid, succinic acid, oxydisuccinic acid, polymaleic acid, benzene, 1,3,5-tricarboxylic acid and carboxymethyloxysuccinic acid and soluble salts thereof. Citrate builders, for example, citric acid and soluble salts thereof (particularly sodium salt) are polycarboxylate builders of particular importance for liquid heavy-duty detergent formulations, but which can also be used in granular compositions . Other carboxylate builders include the carboxylated carbohydrates described in US Pat. 3,723,322, Diehl, issued March 28, 1973 incorporated herein by reference. Also suitable in the detergent compositions of the present invention are the 3,3-dicarboxy-4-oxa-1,6-hexanedioates and the related compounds described in US Pat. 4,566,984, Bush, issued January 28, 1986, incorporated herein by reference. Useful succinic acid builders include C5-C20 alkylsuccinic acid and salts thereof. A particularly preferred compound of this type is dodecenylsuccinic acid. Alkylsuccinic acids are typically of the general formula R-CH (COOH) CH2 (COOH), for example, succinic acid derivatives where R is hydrocarbon, for example C10-C20 alkyl or alkenyl. preferably C 12 -C 16 or where R can be substituted with hydroxyl, sulfo, sulfoxy or sulfon substituents, as described in the patents mentioned above. Succinate builders preferably are used in the form of their water soluble salts, including the sodium, potassium, ammonium and alkanolammonium salts. Specific examples of suctionate detergency builders include: lauryl-succinate, myristylsuccinate, palmitylsuccinate, 2-dodecenylsuccinate (preferred), 2-pentadecenylsuccinate and the like. Lauryl succinate and the preferred builders of this group are described in European Patent Application No. 86200690.5 / 0200263, published November 5, 1986. Examples of useful detergency builders also include sodium and potassium polyacrylates, carboxymethyloxymonalonate , carboxymethyloxysuccinate, cis-cyclohexane hexacarboxylate, cis-cyclopentane tetracarboxylate, water-soluble (these polyacrylates having molecular weights greater than about 2,000 can also be effectively used as dispersants) and the copolymers of maleic anhydride with methylvinyl ether or ethylene . Other suitable polycarboxylates are the polyacetal carboxylates described in US Pat. 4,144,226, Crutchfield, et al., Issued March 13, 1979 incorporated herein by reference. These polyacetal carboxylates can be prepared by joining, under polymerization conditions, a glyoxylic acid ester and a polymerization initiator. The resulting polyacetal carboxylate ether is subsequently attached to the chemically stable end groups to stabilize the polyacetal caboxylate against rapid polymerization in alkaline solution, converted to the corresponding salt and added to the surfactant. Polycarboxylate builders also are described in US Pat. 3,308,067, Diehl, issued March 7, 1967, incorporated herein by reference. Said materials include water-soluble salts of homo and copolymers of aliphatic carboxylic acids such as maleic acid, itaconic acid and methyllenmalonic acid. Other organic builders known in the art can also be used. For example, monocarboxylic acids and soluble salts thereof can be used, which has long chain hydrocarbyl. These would include materials generally referred to as "soaps". Chain lengths of C10-20 are typically used. The hydrocarbyls can be saturated or unsaturated. Other optional ingredients include soil release agents, chelating agents, clay, dirt removal / anti-redeposition agents, polymeric dispersing agents, bleaches, brighteners, suds suppressors, solvents and aesthetic agents. The detergent composition herein can be formulated as a variety of compositions, for example as a laundry detergent, as well as hard surface cleaners or dishwashing compositions. The compositions according to the present invention are further illustrated by the following examples.

EXAMPLE I The following compositions are made by combining the ingredients listed in the listed proportions. 1) N- (methylsulfonyl) -Phe-Gly-Ala-LeuH, which was prepared according to the synthesis of example 2.

EXAMPLE II The following formula is prepared: • Peptide Aldehyde from the synthesis of Example 2 EXAMPLE III The following compositions are made by combining listed in the proportions listed. 1x = the degree of ethoxylation. The average degree of ethoxylation for the compositions are: A = 2.2, B = 0.6, C = 1.4, D = 2.2. 2 The peptide aldehyde of the synthesis of Example 2 is used herein.

Claims (2)

NOVELTY OF THE INVENTION CLAIMS

1- A liquid detergent composition comprising: a) from about 1% to about 95% by weight of the composition of a detersive surfactant; b) a proteolytic enzyme; and c) a protease peptide inhibitor having the formula: Z-B-NH-CH (R) -C (O) -X wherein B is a peptide chain comprising 2 to 5 amino acid portions; X is hydrogen or CF3; Z is an N-blocking portion selected from the group consisting of sulfonamides, re-amonamides, thioureas, sulfinamides, sulphunic acids, sulfunamides, thiocarbamates, trimetic amides, sulphamoyl derivatives and phosphonamides; R is selected from the group consisting of unsubstituted alkyl of C < C6 straight or branched, phenyl and C7-C9 alkylaryl moieties. 2. A liquid detergent composition according to claim 1, further characterized in that R is selected from the group consisting of methyl, iso-propyl, sec-butyl, isobutyl, -CeHs.-C ^ -CßHs, and -CH2CH

2- C6H5 3. A liquid detergent composition according to any of claims 1 or 2 comprising: a) from about 8 to about 70% of said detersive surfactant agent; b) from about 0.0001% to about 5% of a proteolytic enzyme; c) From about 0.00001% to about 5% of a peptide protease inhibitor. 4. A liquid detergent composition according to any of claims 1-3, further characterized in that it comprises a source of calcium ions. 5. A liquid detergent composition according to any of claims 1-4, further characterized in that the N-blocking Z portion is selected from the group consisting of (R "O) 2 (O) p -. (SR") 2-, R "(O) 2S -., SO3H, (R") 2 (0) P, R "O (O) 2S-, (R") 2N (O) C- "R" O (S) C-, R "-P (O) OH and R" O (OH) (O) P-, further characterized in that each R "is independently selected from the group consisting of straight-branched C ^ -CQ -substituted alkyl, phenyl , C7-C9 alkylaryl, and cycloalkyl portions, further characterized in that the cycloalkyl ring may be C4-C8 and may contain one or more heteroatoms selected from the group consisting of O, N, and S. 6.- A detergent composition liquid according to any of claims 1-5, further characterized in that it comprises a source of calcium ion selected from the form of calcium, calcium chloride, calcium acetate, calcium xylene sulfate, calcium sulfate and mixtures thereof. 7. - A liquid detergent composition according to any of claims 1-6, further characterized in that said proteolytic enzyme is a subtilisin type protease. 8. A liquid detergent composition according to claim 7, further characterized in that said proteolytic enzyme is selected from the group consisting of Alcalaza®, Subtilisin BPN ', Protease A, Protease B, and mixtures thereof. 9. A liquid detergent composition according to any of claims 1-8, further characterized by said composition is a light duty detergent composition suitable for dishwashing. 10. A liquid detergent composition for dishwashing according to claim 9, further characterized in that it comprises one or more of the following: foam impellers, chelators, polyacrylate polymers, dispersing agents, dyes, perfumes, processing aids and mixtures thereof. 11. A liquid dishwashing detergent composition according to claim 10, further characterized in that it comprises on top of amylase. 12. A liquid dishwashing detergent composition according to claim 11, further characterized in that it comprises from about 0.25% about 10% by weight of boric acid or a compound layers of forming boric acid or a polyol. 13. - A liquid detergent composition according to any of claims 1-8, further characterized in that said composition is a heavy duty detergent composition suitable for laundry washing. 14. A liquid laundry detergent composition according to claim 13, further characterized in that it comprises an effective amount of one or more of the following secondary enzymes: lipase, amylase, cellulase and mixtures thereof. 15. A liquid laundry detergent composition according to claim 14, further characterized in that it comprises one or more of the following: foam impellers, detergent builders, soil release polymers, polyacrylate polymers, dispersing agents, inhibitors of dye transfer, dyes, pertumes, processing aids, brighteners and mixtures thereof. 16. A liquid laundry detergent composition according to claim 14, further characterized in that said second enzyme is lipase. 17. A liquid laundry detergent composition according to claim 16, further characterized in that the lipase is obtained by cloning the gene in Humicola Lanuginosa and expressing the gene in Aspergillus Orvzae. 18. - A composition according to claim 14, further characterized in that said second over is a cellulase derived from Humicola Insolens and further characterized in that said composition comprises from 0.0001% to 0.1% by weight of the total composition of said cellulase. 19. A liquid laundry detergent composition according to claim 13, further characterized in that it comprises from about 0.25% to about 10% by weight of boric acid or a compound layers of forming boric acid and a polyol. 20.- The peptide compounds CH3S? 2Phe-Gly-Ala-Leu-H, CH3SO2Val-Ala-Leu-H, C6H5CH2O (OH) (O) P-Val-Ala-Leu-H, C6H5CH2O (OH) (O) P-Val-Ala-Leu-CF3, CH3CH2SO2-Phe-Gly-Ala-Leu-H, C6H5CH2SO-Val-Ala-Leu-H, C6H5CH2O (OH) (O) P-Leu Ala-Leu-H, C 6 H 5 CH 2 O (OH) (O) P-Phe-Ala-Leu- H and Ch 3 O (OH) (O) P-Leu-Gley-Ala-Leu-H.