MXPA00003978A

MXPA00003978A - Cleaning compositions containing multiply-substituted protease variants

Info

Publication number: MXPA00003978A
Application number: MXPA/A/2000/003978A
Authority: MX
Inventors: Andre Cesar Baeck; Alfred Busch; Michael Stanford Showell; Chanchal Kumar Ghosh; Ryohei Ohtani; Volker Schellenberger; James T Kellis Jr; Christian Paech; Joanne Nadherny; Donald P Naki; Katherine D Collier; Robert M Caldwell; Ayrookaran J Poulose
Original assignee: The Procter & Gamble Company
Priority date: 1997-10-23
Filing date: 2000-04-24
Publication date: 2001-06-26

Abstract

The present invention relates to cleaning compositions comprising a protease variant. One cleaning composition comprises a protease variant including a substitution of an amino acid residue with another naturally occurring amino acid residue at an amino acid residue position corresponding to position 103 of i(Bacillus amyloliquefaciens) subtilisin in combination with a substitution of an amino acid residue with another naturally occurring amino acid residue at one or more amino acid residue positions corresponding to positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 111, 114, 116, 117, 119, 121, 123, 126, 128,130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243,244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of i(Bacillus amyloliquefaciens) subtilisin;wherein when said protease variant includes a substitution of amino acid residues positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions other than amino acid residue positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of i(Bacillus amyloliquefaciens) subtilisin;and one or more cleaning adjunct materials. Another cleaning composition comprises a protease variant including a substitution of an amino acid residue with another naturally occurring amino acid residue at one or more amino acid residue positions corresponding to positions 62, 212, 230, 232, 252 and 257 of$(Bacillus amyloliquefaciens) subtilisin;and one or more cleaning adjunct materials. Methods for using the cleaning compositions are also provided.

Description

CLEANING COMPOSITIONS COMPRISING MULTIPLE-REPLICED PROTEASE VARIANTS FIELD OF THE INVENTION The present invention relates to cleaning compositions comprising one or more protease variants with multiple substitutions and one or more cleaning adjunct materials. Most particularly, the present invention relates to laundry detergent compositions, dishwashing detergent compositions, hard surface cleaning compositions and personal cleansing compositions comprising one or more protease variants with multiple substitutions and one or more adjunct materials. cleaning.

BACKGROUND OF THE INVENTION Several types of enzymes have been used in laundry detergents for a long time to aid in the removal of certain stains on fabrics. Each kind of enzyme (amylase, protease, etc.) generally catalyzes a different chemical reaction. For example, protease enzymes are known for their ability to hydrolyze (decompose a compound into two or more simple compounds) of other proteins. This ability has had great advantage through the incorporation of protease enzymes that occur naturally or that are genetically engineered for laundry detergent compositions. In recent years, the use of enzymes for use in automatic dishwashing compositions has also been investigated. Unfortunately, many enzymes, such as various conventional protease enzymes, are not transferred properly to the wash environment. Specifically, thermal stability, pH stability, oxidative stability and the specific character of the substrate need to be optimized to ensure satisfactory performance. The patent E.U. No. RE 34,606 to Estell et al. describes the modification of amino acid residues of subtilisin corresponding to the positions in Bacillus amyloliquefaciens of subtilisin tyrosine-1, aspartate + 32, aspargin + 55, tyrosine + 104, methionine + 222, glycine + 166, histidine + 64, glycine + 169, phenylalanine + 189, serine + 33, serine + 221, tyrosine + 217, glutamate + 156 and alanine + 152. EU patent No. 5,182,204 describes the modification of the residue +224 amino acid in subtilisin of Bacillus amyloliquefaciens and the equivalent positions in other subtilisins that can be modified by substitution, insertion or deletion and that can be combined with modifications to the residues identified in the patent E.U. No. RE 34,606 to form useful subtilisin mutants or variants. The patent E.U. No. 5,182,204 further describes the modification of several amino acid residues within subtilisin, specifically including +99, +101, +103, +107, +126, +128, +135, +197 and +204. The patent E.U. No. 5,155,033 describes similar mutant subtilisins that have a modification at a position equivalent to +225 subtilisin of B. amyloliquefaciens. The patent E.U. Nos. 5,185,258 and 5,204,015 describe mutant subtilisins that have a modification at positions +123 and / or +274. The patent E.U. No. 4,914,031 discloses certain analogues to subtilisin, including a subtilisin modified at position +76. The patent E.U. No. 5,679,630 to Baeck et al. describes cleaning compositions comprising a protease variant that includes substitutions of amino acid residues with other amino acid residues at positions corresponding to position 76 in combination with one or more of the following positions 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 of Bacillus amyloliquefaciens subtilisin, and one or more cleaning composition materials. However, there still remains a need for proteases, particularly serine proteases, which provide enhanced cleaning ability when used in detergent and cleaning compositions. In addition, the specific combinations claimed in the present application are not identified in any of these references of the prior art.

BRIEF DESCRIPTION OF THE INVENTION The present invention fulfills the aforementioned needs in the fact that it has surprisingly been discovered that the protease variants with multiple substitutions of the present invention, when used in cleaning compositions, provide the improved and increased cleaning ability, including, but not limiting, the removal of stains and / or dirt and / or reduction and / or maintenance of whiteness and / or cleaning of dirt and / or stains and / or removal and / or reduction of films on conventional cleaning compositions containing protease . The protease variants with multiple substitutions of the present invention are suitable for use in high and low density granular compositions, heavy and light duty liquids, tablets, as well as synthetic bar detergent compositions, and other cleaning compositions. In one aspect of the present invention, a cleaning composition comprises: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0. 001% to about 10% by weight of the cleaning composition of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another amino acid residue that occurs naturally to a residue position of amino acids corresponding to position 103 of subtilisin Bacillus amyloliquefaciens in combination with a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 74, 75, 76, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 1 1 1, 1 14, 116, 1 17, 119, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209 , 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of subtilisin from Bacillus amyloliquefaciens; wherein each protease variant includes a substitution of amino acid residues at positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the amino acid residue positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and b) one or more cleaning attachment materials. In still another aspect of the present invention, a fabric cleaning composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.0001% to about 10% by weight of the fabric cleaning composition of a protease variant, wherein said protease variant is described above; b) at least about 5% by weight of the fabric cleaning composition of a surfactant; and c) at least about 5% by weight of the fabric cleaning composition of a builder is provided. In yet another aspect of the present invention, there is provided a method for cleaning a fabric that requires cleaning, which comprises contacting the fabric with the fabric cleaning composition of the present invention. In yet another aspect of the present invention, a dishwashing composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.0001% to about 10% by weight of the dishwashing composition of a protease variant, wherein said protease variant was described above; and b) about 0.1% to about 10% by weight of a surfactant agent is provided.

In yet another aspect of the invention, there is provided a method for cleaning a dish that requires cleaning, which comprises contacting the piece of dishware with the dishwashing composition of the present invention. In still another aspect of the present invention, a personal cleansing composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.001% to about 5% by weight of the composition of personal cleansing of a protease variant, wherein said protease was described above; b) from about 0.1% to about 95% by weight of a surfactant system, for a personal cleansing composition; c) optionally, from about 0.05% to about 50% by weight of the personal cleansing composition of an enzyme stabilizer is provided. Still in another embodiment of the present invention, a method is provided for the personal cleansing of a part of the human being or the body of a lower animal that requires the need for cleaning; method comprising contacting said part with the personal cleansing composition of the present invention. In still another aspect of the present invention, a cleaning composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.0001% to about 10% by weight of the composition of cleaning a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 62, 212, 230 , 232, 252, and 257 of Bacillus amyloliquefaciens subtilisin; and b) one or more cleaning attachment materials are provided. In yet another aspect of the present invention, a fabric cleaning composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.0001% to about 10% by weight of the fabric cleaning composition of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 62 , 212, 230, 232, 252 and 257 of Bacillus amyloliquefaciens subtilisin; b) at least about 5% by weight of the fabric cleaning composition, of a surfactant; and c) at least about 5% by weight of the fabric cleaning composition, of a builder, is provided. Still in still another aspect of the present invention, there is provided a method for cleaning fabrics that require cleaning, which consists in contacting the fabric with the fabric cleaning composition of the present invention. Yet another aspect of the present invention, a dishwashing composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.0001% to about 10% by weight of the composition fabric cleaning of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 62 , 212, 230, 232, 252 and 257 of Bacillus amyloliquefaciens subtilisin; and b) about 0.1% to about 10% by weight of the dishwashing composition of a surfactant is provided. Still another embodiment or aspect of the present invention provides a method for cleaning dishes that require cleaning. , which consists in contacting the piece of tableware with the dishwashing composition of the present invention. Still in another aspect of the present invention, a personal cleansing composition comprising: a) a protease variant, preferably an effective amount of a protease variant, most preferably from about 0.001% to about 5% by weight of the personal cleansing composition of a protease variant , wherein said protease variant includes a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 62, 212, 230, 232, 252 and 257 of subtilisin from Bacillus amyloliquefaciens; and b) from about 0.1% to about 95% by weight of the personal cleansing composition, of a surfactant; and c) optionally, from about 0.05% to about 50% by weight of the personal cleansing composition, of an enzyme stabilizer is provided. Still another embodiment of the present invention, there is provided a method for personal cleansing of a part of a human being or a body of a lower animal in need of cleaning, which comprises contacting said part with the personal cleansing composition of the present invention. Accordingly, it is an object of the present invention to provide cleaning compositions having a protease variant capable of providing improved and increased cleaning of fabrics, tableware, tableware, kitchenware, kitchenware and other hard surface substrates. Still another object of the present invention is to provide methods for cleaning fabrics, tableware, tableware, kitchenware, kitchenware and other hard surface substrates by using cleaning compositions containing a protease variant of the present invention. These and other objects, features and advantages will be apparent from the following detailed description, examples and appended claims. All percentages, ratios and ratios herein are based on weight, unless otherwise indicated. All documents cited herein are incorporated therein by reference.

BRIEF DESCRIPTION OF THE DRAWINGS Figures 1A-C describe the DNA and amino acid sequence for the subtilisin Bacillus amyloliquefaciens and a partial restriction map of this gene. Figure 2 describes the amino acid residues conserved between the subtilisins from Bacillus amyloliquefaciens (BPN) 'and Bacillus lentus (wild type). Figures 3A and 3B describe the amino acid sequence of four subtilisins. The upper line represents the amino acid sequence of subtilisin from subtilisin Bacillus amyloliquefaciens (which is sometimes referred to as subtilisin BPN '). The second line describes the amino acid sequence of subtilisin from Bacillus subtilis. The third line describes the amino acid sequence of subtilisin from ß. licheniformis. The fourth line describes the amino acid sequence of subtilisin from Bacillus lentus (also referred to as subtilisin 309 in PCT WO89 / 06276. The symbol of * denotes the absence of specific amino acid residues compared to subtilisin BPN '.

DETAILED DESCRIPTION OF THE INVENTION Proteases Proteases are carbonylhydrolases that generally act to cut peptide or protein peptide bonds. As used herein, "protease" means a protease that occurs naturally or recombinant protease. Naturally occurring proteases include a-aminoacylpeptide hydrolase, peptidylamino acid hydrolase, acylamino hydrolase, serine carboxypeptidase, metallocarboxypeptidase, thiolproteinase, carboxylproteinase, and metalloproteinase. Serinoproteases, metalloproteases, thiol proteases as well as acid proteases are included as well as endo and exo proteases. The present invention includes protease enzymes that do not occur naturally, which are variants of carbonyl hydrolase (protease variants) having different proteolytic activity, stability, substrate specificity, pH profile and / or performance characteristics compared with the precursor carbonyl hydrolase from which the amino acid sequence variant is derived. Specifically, said protease variants have an amino acid sequence not found in nature, which is derived by the replacement of a plurality of amino acid residues of a precursor protease with different amino acids. The precursor protease can be a protease that occurs naturally or recombinant protease. As stated above, the protease variants are designed to have a specific character in the form of trypsin and are preferably also stable to bleaching. The protease variants useful herein encompass the substitution of any of the positions of the nineteen positions of L-amino acids that occur naturally at the designated amino acid residue positions. Said substitutions can be made in any precursor subtilisin (prokaryotic, eukaryotic, mammalian, etc.). Throughout this application reference several amino acids are specified by common one to three letter codes. These codes are identified in Dale, M.W. (1989), Molecular Genetics of Battery, John Wiley & Sons, Ltd., appendix B. The protease variants useful herein are preferably derived from a Bacillus subtilisin. Most preferably, protease variants are derived from subtilisin Bacillus lentus and / or subtilisin 309.

Carbonyl hydrolases Carbonyl hydrolases are protease enzymes that hydrolyze compounds that contain O II C-X links where X is oxygen or nitrogen. They include carbonyl hydrolases that occur naturally and recombinant carbonyl hydrolases. Naturally occurring carbonyl hydrolases mainly include hydrolases, for example, peptide hydrolases such as subtilisins or metalloproteases. Peptide hydrolases include hydrolase of -aminoacylpeptide, hydrolase of peptidylamino acid, acylamino hydrolase, serine carboxypeptidase, metallocarboxypeptidase, thiol proteinase, carboxylproteinase and metalloproteinase. Serinoproteases, metalloproteases, thiol proteases and acid proteases are included, as well as endo and exo-proteases. Subtilisins Subtilisins are bacterial or fungal proteases that generally act to cut the peptide bonds of proteins or peptides. As used herein, "subtilisin" means a subtilisin that occurs naturally or a recombinant subtilisin. It is known that a series of subtilisins that occur naturally are produced and are often secreted by several microbial species. The amino acid sequences of the members of these series are not fully homologous. However, the subtilisins in these series exhibit the same or similar type of proteolytic activity. This class of serine proteases share a common amino acid sequence that defines a trivalent catalytic radical that distinguishes them from chymotrypsin related to serine proteases. Subtilisins and serine proteases related to chymotrypsin both have a trivalent catalytic radical comprising aspartate, histidine and serine. In proteases related to subtilisin, the relative order of these amino acids, reading from amino to carboxy terminals, is aspartate-histidine-serine. However, in proteases related to chymotrypsin, the relative order is histidine-aspartate-serine. Therefore, subtilisin herein refers to a serine protease having the trivalent catalytic radical of proteases related to subtilisin. Examples thereof include, but are not limited to, the subtilisins identified in Figure 3 herein. Generally, and for purposes of the present invention, the numbering of the amino acids in the proteases corresponds to the numbers assigned to the mature subtilisin Bacillus amyloliquefaciens which is the sequence presented in Figure 1. Protease variants A "protease variant" has an amino acid sequence that is derived from the amino acid sequence of a "precursor protease". The precursor proteases include proteases that occur naturally and recombinant proteases. The amino acid sequence of the protease variant is "derived" from the amino acid sequence precursor protease by substitution, deletion or insertion of one or more amino acids of the precursor amino acid sequence. Said modification is of the "precursor DNA sequence" which codes for the amino acid sequence of the precursor protease instead of the manipulation of the precursor protease enzyme per se. Suitable methods for such manipulation of the DNA precursor sequence are described which includes methods that are described herein, as well as methods known to those skilled in the art (see for example, EP 0 328 299, WO 89/06279 and patents and US applications defined herein). In a preferred embodiment, the protease variants that are protease enzymes useful in the present invention of the cleaning compositions herein comprise protease variants including a substitution of an amino acid residue with another amino acid residue that occurs naturally at an amino acid residue position corresponding to position 103 of subtilisin Bacillus amyloliquefaciens in combination with a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to the positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 1 1 1, 1 14 , 1 16, 1 17, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227 , 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260 , 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of subtilisin Bacillus amyloliquefaciens; wherein said protease variant includes a substitution of amino acid residues at the positions corresponding to positions 103 and 76, where there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the residue positions of amino acids corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and one or more cleaning attachment materials. Although any combination of the above list of amino acid substitutions may be employed, the preferred protease variants enzymes useful for the present invention comprise substitution, deletion or insertion of amino acid residues in the following combinations: 1) a protease variant that includes substitutions of amino acid residues at position 103 and at one or more of the following positions 236 and 245; 2) a protease variant that includes substitutions of amino acid residues at positions 103 and 236 and at one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104 , 109, 130, 131, 159, 183, 185, 205, 209, 210, 211, 212, 213, 215, 217, 230, 232, 248, 252, 257, 260, 270 and 275; 3) a protease variant that includes substitutions of amino acid residues at positions 103 and 245 and in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104 , 109, 130, 131, 159, 170, 183, 185, 205, 209, 210, 21 1, 212, 213, 215, 217, 222, 230, 232, 248, 252, 257, 260, 261, 270 and 275; and 4) a protease variant that includes substitutions of the amino acid residues at positions 103, 236 and 245 and in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 21 1, 212, 213, 215, 217, 230, 232, 243, 248, 252, 257, 260, 270 and 275. A more preferred protease variant useful in the cleansing compositions of the present invention includes a substitution group (a substitution group per row in the following Table I) selected from group consisting of: TABLE I A more preferred protease variant, useful in the cleansing compositions of the present invention, includes a group of substitutions (a substitution group per row in the following Table II) selected from a group consisting of: TABLE II A still more preferred protease variant useful in the composition of the present invention includes a substitution group selected from the group consisting of the substitution groups of Table 1, except for the following substitution groups in Table III: TABLE III A still highly preferred protease variant, useful in the composition of the present invention includes a substitution group selected from the group consisting of the substitution groups of Table IV: Still an even more preferred protease variant, useful in the composition of the present invention includes a substitution group selected from the group consisting of the substitution groups of the V: TABLE V A highly preferred protease variant useful in the cleansing compositions of the present invention includes a substitution group selected from the group consisting of: 12/102/103/104/159/212/232/236/245/248/252; 12/76/103/104/130/170/185/222/243/245; 12/76/103/104/130/222/245/261; 12/76/103/104/130/222/245; 12/76/103/104/222/245; 61/68/103/104/159/232 236/245/248/252; 62/103/104/159/213/232/236/245/248/252 62/103/104/109/159/213/232/236/245/248/252; 62/103/104/159/232/236/245/248/252; 62/101/103/104/159/212/213/232/236 245/248/252; 62/103/104/130/159/213/232/236 245/248/252; 68/103/104/159/232/236/245/248/252 270: 68/103/104/159/185/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/213/232/236/245/248/252 68/103/104/159/230 232/236/245; 68/76/103/104/159/209/232/236/245; 68/103/104/232/236/245/248/257/275; 68/103/104/213/232/236/245 248/252; 68/103/104/159/232/236/245/248/252; 68/103/104/159/209/232/236/245; 68/76/103/104/159/236; 68/76/103/104 / 159/236/245; 68/76/103/104/159/232/236/245; 68/103/104/159/232/236/245/252; 68/103/104/159/232/236/245; 68/103/104/159/232/236/245/257; 68/76/103/104/159/211/232/236/245; 68/76/103/104/159/215/232/236/245; 68/103/104/159/210/232/236/245; 68/103/104/159/213/232/236/245/260; 68/76/103/104/159/213/232/236/245/260; 68/103/104/159/236; 68/76/103/104/159/210/2327236/245/260; 68/103/104/159/236/245; 68/103/104/159/183/232/236/245/248/252; 68/76/103/104/159/236/245; 68/103/104/232/236/245/257/275; 68/103/104/159/213/232/236/245; 76/103/222/245; 76/103/104/222/245; 76/103/104/159/232/236/245; 76/103/104-'159 / 213/232/236/245/260; 76/103/104/159: 76/103 / 104''131 / 159/232/236/245/248/252; 97/103/104/159/232/236/245/248/252; 98/102 103 / ¡04/159/212/232/236/245/248/252; 98/103/104/159/232/236/245/248/252; 101/103/104/159/232/236/245/248/252; 102/103/104/159/232/236/245/248/252; 103/104/159/232/236/245; 103/104 / 159/232/236/245/248/252, 103/104/159/205/209/232/236/245/257 103/104/159/232/245/248/252; 103/104/159/205/209/210/232/236/245/257; 103/104/159/213/232/236/245/248/252; 103/104/159/217/232/236/245/248/252: 103/104/130/159/232/236/245/248/252; 103/104/159/230/236/245; 103/104/159/236/245; 103/104/159/248/252/270; 103/104/13 1/159/232/236/245/248/252; 103/104/159/205/209/232/236/245; and 103/104/159/232/236/245/257.

A still more preferred protease variant useful in the cleansing compositions of the present invention includes a substitution group selected from the group consisting of: 12R / 76D / 103A / 104T / 130T / 222S / 245R; 12R / 76D / 103A / I04I / 222S / 245R; 12R / 102A / 103A / 104I / 159D / 212G / 232V / 236H / 245R / 248D / 252K; i 2R / 76D / 103 A / 104T / 130G / 222S / 245R / 261 D; 12R / 76D / 103A / 104T / 130G / 170S / 185D / 222S / 243D / 245R; 61E / 68A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252K; 62D / 103A / 104I / 109R / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 62D / 103A / 104I / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 62D / 103 A / 1041 / 159D / 232V / 236H / 245R / 248D / 252; 62D / 103A / 104I / 130G / 159D / 213R 232V / 236H / 245R 248D / 252K; 62D / 10JG / 103A / 104I / 159D / 212G / 213R / 232V / 236H / 245R / 248D / 252K; "" 68A / 103A / 104I / I59D / 232V / 236H 245R / 248D / 252K 270A; 68A 76D / 103 A / 1041 / 159D / 213R / 232 V / 236H 245R 260A; 68A / 103A / 1041 / 159D / 236H; 68A / 103A 104I / 159D / 236H / 245R; 68A / 76D / 103 A / 1041 / 159D / 210I / 232V / 236H / 245R / 260A; 68A / 103A 104I / 159D / 183D / 232V / 236H / 245R / 248D / 252K; 68A 103A 104I / 159D / 209W / 232V / 236H / 245R; 68A / 76D / 103A / 1041 / 159D / 211 R / 232V / 236H / 245R; 68A / 76D / 103 A / 1041 / 159D / 215R / 232 V / 236H / 245R; 68A / 103 A / 1041 / 159D / 2 and 3R / 232 V / 236H / 245R / 260A; 68 A 76 D / 03 A 1041/159 D / 236 H: 68 A 76 D / 103 A 104 I / 159 D / 236 H 245 R; 68A / 76D / 103A 104I / 159D / 232V / 236H / 245R: 68A / 103 A 1041 / 159D / 232V / 236H 245R / 252K; 68A / 103A 1041 / 159D / 232V / 236H / 245R; 68A / 103 A / 1041 / 159D / 232V / 236H / 245R 257V; 68A 103A 1041 / 159D / 185D / 232V / 236H / 245R 248D / 252K; 68 A 103 A / 1041 / 159D / 210L / 232V / 236H / 245R / 248D / 252K; A / 103 A / 1041 / 159D / 185D / 210L / 232 V / 236H 245R / 248D / 252K; 68A / 103A / 1041 / 159D / 213E / 232V / 236H / 245R / 248D / 252; . 68 A / 103 A 1041 / 159D / 230 V / 232V / 236H / 245R; 68A / 76D / 103 A / 1041 / 159D / 209W / 232V / 236H 245R; 68A / 103A / 104I / 232V / 236H / 245R / 248D / 257V / 275H; 68A / 103A / 104I / 232V / 236H 245R / 257V / 275H; 68A / 103 A / 1041 / 213E / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 210I / 232V / 236H / 245R; 68A 103A / 104I / 159D / 210L / 232V / 236H / 245R; 68A / 103A / 1041 / 159D / 213G / 232V / 236H / 245R; 76D / 103A / 222S / 245R; 76D / 103A 104I / 222S / 245R; 76D / 103A / 104I / 159D / 232V / 236H / 245R; 76D / 103 A / 1041 / 159D; 76D / 103A / 104I / 131V / 159D / 232V / 236H / 245R 248D / 252K; 76D / 103A / 104I / 159D / 213R / 232V / 236H / 245R / 260A; 97E / 103 A / 1041 159D / 232V / 236H / 245R / 248D / 252K; 98L / 103A 104yi59D / 232V / 236H / 245R / 248D / 252K; 8L / 102A / 103 A / 1041 / 159D / 212G / 232V / 236H / 245R / 248D / 252K; 101G / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252K; 102A / 103A 104I / 159D / 232V / 236H / 245R / 248D / 252K; 103A / 104I / I59D / 232V / 236H / 245R 248D / 252K; 103A / 104I / 159D / 213R / 232V / 236H 245R / 248D / 252; 103A / 104I / 130G / 159D / 232V / 236H 245R / 248D / 252K; 103A / 104I / 159D / 230V / 236H / 245R; 103 A / 1041 / 159D / 217E / 232 V / 236H / 245R / 248D / 252K; 103 A / 1041 / 159D / 236H / 245R; 103A / 1041 / 159D / 248D / 252K 270V; 103 A / 1041 / 159D / 232V / 236H / 245R; 103 A / 1041 / 159D / 205I / 209W / 232V / 236H / 245R; 103A / 1041 / 159D / 232V / 236H / 245R / 257V: 103 A 1041 / 159D / 205I / 209W / 232V / 236H 245R 257V; 103 A 1041/131 V / l 59D / 232V / 236H / 245R / 248D / 252K; 103A 104I / 159D / 205I / 209W / 210I / 232V / 236H 245R 257V; and 103 A 1041 / 159D / 232V / 245R7248D / 252K.

A yet even more preferred protease variant useful in the cleaning compositions of the present invention includes a substitution group selected from the group consisting of: 12/76/103/104/130/222/245/261; 62/103/104/159/232/236/245/248/252; 62/103/104/159/213/232/236/245/248/252; 62/101/103/104/159/212/213/232/236/245/248/252; 68/103/104/159/232/236/245; 68/103/104/159/230/232/236/245; 68/103/104/159/209/232/236/245; 68/103/104/159/232/236/245/257 68/76/103/104/159/213/232/236/245/260; 68/103/104/159/213/232/236/245/248/252; 68/103/104/159/183/232/236/245/248/252; 68/103/104/159/185/232/236/245/248/252; 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252; 68/103/104/159/213/232/236/245; 98/103/104/159/232/236/245/248/252; 98/102/103/104/159/212/232/236/245/248/252; 101/103/104/159/232/236/245/248/252; 102/103/104/159/232/236/245/248/252; 103/104/159/230/236/245; 103/104/159/232/236/245/248/252; 103/104/159/217/232/236/245/248/252; 103/104/130/159/232/236/245/248/252; 103/104/131/159/232/236/245/248/252; 103/104/159/213/232/236/245/248/252; and 103/104/159/232/236/245 The even more preferred protease variant, useful in the cleansing compositions of the present invention, includes a substitution group selected from the group consisting of: 12R / 76D / 103 A 104T / 130T / 222S / 245R 261 D: 62D / 103A / 104I / 159D / 232V / 236H 245R / 248D / 252K; 62D / 103A 1041 / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 1041 / 159D / 209W / 232V / 236H 245R; 68 A 76D / 103 A / 1041 / 159D / 213R / 232V / 236H / 245R / 260 A; 68A / 103A / 104I / 159D / 213E / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 183D / 232V / 236H / 245R / 248D / 252; 68A 103 A / 1041 / 159D / 232V / 236H / 245R; 68A / 103A 104I / 159D / 230V / 232V / 236H / 245R; 68A / 103A 104I / 159D / 232V / 236H / 245R / 257V; 68A / 103 A / 1041 / 159D / 213G / 232V / 236H / 245R / 248D / 252; • 68A / 103A / 104I / I59D / 185D / 232V / 236H / 245R 248D / 252K; 68A / 103 A / 1041 / 159D / 185D / 210L / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 210L / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 1041 / 159D / 213G / 232V / 236H / 245R; 98L / 103 A 1041 / 159D / 232V / 236H / 245R 248D / 252K; 98L / 102A 103A / 1041 / 159D / 212G / 232V / 236H 245R / 248D / 252K; 101G / 103A 104I / 159D / 232V / 236H / 245R / 248D / 252K; 102A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252; 103 A 1041 / 159D / 230V / 236H 245R; 103A 104I / 159D / 232V / 236H 245R / 248D / 252K; 103 A / 1041 / 159D / 217E / 232V / 236H / 245R / 248D / 252K; 103A / 104I / 130G / 159D / 232V / 236H / 245R / 248D / 252K; 103A / I04I / 131V / 159D / 232V / 236H / 245R / 248D / 252K; 103jV104I / 159D / 213R 232V / 236H 245R / 248D / 252K; and 103A / 104I / 159D / 232V / 236H 245R.

In another preferred embodiment, protease variants that are protease enzymes useful in the cleansing compositions of the present invention comprise protease variants that include a substitution of an amino acid residue with another amino acid residue that occurs naturally, which correspond to positions 62,212,230, 232, 252 and 257 of subtilisin Bacillus amyloliquefaciens. Although any combination of the above list of amino acid substitutions may be employed, the preferred protease variant enzymes useful for the present invention comprise substitution, deletion or insertion of amino acid residues in the following combinations: 1) a protease variant including substitutions of amino acid residues at position 62 and one or more of the following positions: 103, 104, 109, 159, 213, 232, 236, 245, 248 and 252; 2) a protease variant including substitutions of the amino acid residues at position 212 and one or more of the following positions: 12, 98, 102, 103, 104, 159, 232, 236, 245, 248 and 252; 3) a protease variant including substitutions of the amino acid residues at position 230 and one or more of the following positions: 68, 103, 104, 159, 232, 236 and 245; 4) a protease variant including substitutions of the amino acid residues at position 232 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 5) a protease variant including substitutions of amino acid residues at position 232 and one or more of the following positions: 103, 104, 236 and 245; 6) a protease variant including substitutions of amino acid residues at positions 232 and 103 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 7) a protease variant including substitutions of the amino acid residues at positions 232 and 104 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 8) a protease variant including substitutions of the amino acid residues at position 232 and 236 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 9) a protease variant including substitutions of the amino acid residues at position 232 and 245 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 10) a protease variant including substitutions of the amino acid residues at position 232, 103, 104, 236 and 245 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270, and 275; 1 1) a protease variant including substitutions of the amino acid residues at position 252 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109 , 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248, 270; 12) a protease variant including substitutions of the amino acid residues at position 252 and one or more of the following positions: 103, 104, 236 and 245; 13) a protease variant including substitutions of the amino acid residues at position 252 and 103 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248, and 270; 14) a protease variant including substitutions of the amino acid residues at position 252 and 104 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 15) a protease variant including substitutions of the amino acid residues at position 252 and 236 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 16) a protease variant including substitutions of the amino acid residues at position 252 and 245 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 17) a protease variant including substitutions of the amino acid residues at position 252, 103, 104, 236 and 245 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; and 18) a protease variant including substitutions of the amino acid residues at position 257 and one or more of the following positions: 68, 103, 104, 205, 209, 210, 232, 236, 245 and 275. A variant of Most preferred useful protease in the cleaning compositions of the present invention includes a substitution group (a substitution group per row in the following Table VI) selected from the group consisting of: TABLE VI A still more preferred protease variant, useful in the cleaning compositions of the present invention, includes a substitution group (one substitution per row in the following Table VII), selected from the group consisting of: TABLE VII A still more preferred protease variant, useful in the cleansing composition of the present invention, includes a substitution group, selected from the group consisting of the substitution groups in Table VI except for the following replacement group of the HIV Table. : HIV TABLE A still more preferred protease variant, useful in the cleaning composition of the present invention, includes a substitution group, selected from the group consisting of the substitution groups in Table IX: TABLE IX A still more preferred protease variant, useful in the cleaning composition of the present invention, includes a substitution group, selected from the group consisting of the substitution groups in Table X: TABLE X A still more preferred protease variant, useful in the cleansing compositions of the present invention, includes a substitution group, selected from the group consisting of: 12/102 103/104/159/212/232/236/245/248/252; 61/68/103/104/159/232/236/245/248/252; 62/103/104/130/159/213/232/236/245/248/252; 62/103/104/159/213/232/236/245/248/252; 62/103/104/109/159/213/232/236/245/248/252; 62/103/104/159/232/236/245/248/252; 62/101 / 103/104/159/212/213/232/236/245/248/252; 68/103 / 104/159/232/236/245/248/252/270; 68/103/104/159/185/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252; 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/213/232/236/245/248/252; 68/103/104/159/230/232/236/245; 68/76/103/104/159/209/232/236/245; 68/103/104/232/236/245/248/257/275; 68/103/104/213/232/236/245/248/252; 68/103/104/159/232/236/245/248/252; 68/103/104/159/209/232/236/245; 68/76/103/104/159/232/236/245; 68/103/104/159/232/236/245/252; 68/103/104/159/232/236/245; 68/103/104/159/232/236/245/257; 68/76/1037104/159/21 1/232/236/245; 68/76/103/104/159/215/232/236/245; 68/103/104/159/210/232/236/245; 68/103/104 / 159/213/232/236/245/260; 68/76/103/104/159/213/232/236/245/260; 68/76/103/104/159/210/232/236/245/260; 68/103/104 / 59/183/232/236/245/248/252; 68/103/104/232/236/245/257/275; 68/103/104/159/213/232/236/245; 76/103/104/159/232/236/245; 76/103/104/159/213/232/236/245/260; 76/103/104/131/159/232/236/245/248/252; 97/103/104/159/232/236/245/248/252; 98/103/104/159/232/236/245/248/252; 98/102/103/104/159/212/232/236/245/248/252; 101/103/104/159/232/236/245/248/252; 102/103/104/159/232/236/245/248/252; 103/104/159/232/236/245; 103/104/159/248/252/270; 103/104/159/232/236/245/248/252: 103/104/159/205/209/232/236/245/257 103/104/159/232/245/248/252; 103/104/159/205/209/210/232/236/245/257; 103/104/159 / 13/232/236/245/248/252; 103 /! 04 / 159/217/232/236/245/248/252; 103/104/130/159/232/236/245/248/252; 103/104/13 1/159/232/236/245/248/252; 103/104/159/205/209/232/236/245; and 103/104/159/232/236/245/257.

A very highly preferred protease variant, useful in the cleansing compositions of the present invention, includes a substitution group, selected from the group consisting of: R / 102A 103A / 104I / 159D / 212G / 232V / 236H / 245R / 248D / 252K; 61 E / 68A / 103 A 1041 / 159D / 232V / 236H / 245R / 248D / 252K; D / 103A 1041 / 109R 159D / 213R7232V / 236H245R248D / 252K; 62D / 103A 1041 / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 62D / 103A 1041 / 159D / 232V / 236H / 245R / 248D / 252; D / 103A / 104I / 130G / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 101G / 103A / 1041 / 159D / 212G / 213R / 232V / 236H / 245R / 248D / 252; 68A76D / 103 A / 1041 / 159D / 213R232V / 236H245R260A; 68A / 76D / 103 A / 1041 / 159D / 210I / 232V / 236H / 245R / 260A; 68A 103A / 104I / 159D / 183D / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 209W / 232V / 236H / 245R; 68A / 76D / 103 A / 1041 / 159D / 211 R / 232V / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 215R / 232V / 236H / 245R; 68A 103A / I04I / 159D / 213R / 232V / 236H / 245R260A; 68A / 76D / 103A 104I / 159D / 232V / 236H245R; 68A103A / 104? / 159D / 232V / 236H245R252K; 68A / 103A / 1041 / 159D / 232V / 236H / 245R; 68A / 03 A / 1 41 / 159D / 232V / 236H / 245R / 257V; 68A / 103A / 104I / 159D / 185D / 232V / 236H / 245R / 248D / 252K; 68A / 103 to 1041 / 159D / 2101J232V / 236H245R248D252K; A / 103 A / 1041 / 159D / 185D / 210L / 232V / 236H / 245R / 248D / 252K; 68A 103A / 1041 / 159D / 213E / 232V / 236H / 245R / 248D / 252K; 68A / 103 A / 1041 / 159D / 230V / 232V / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 209W / 232V / 236H245R; 68A / 103A / 104I / 232V / 236H / 245R / 248D / 257V / 275H; 68A 103A 104I / 232V / 236H245R257V / 275H; 68A 103A / 104I / 213E / 232V / 236H / 245R248D / 252K; 68A 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 68A / 103A 104I / 159D / 210I / 232V / 236H245R; 68A / 103 A / 104? / 159D / 210L / 232V / 236H / 245R; 68A / 103 A 1041 / 159D / 213G / 232V / 236H / 245R; Ó8A 103 A 1041 / 159D / 232V / 236H / 245R / 248D / 252K270A; 76D / 103 to 1041 / 159D / 232V / 236H / 245R; 76D / 103A / 04I / Í31V / 159D / 232V / 236H / 245R / 248D / 252K; 76D / 103A 104I / 159D / 213R232V / 236H / 245R / 260A; 97E / 103A / 104I / 159D / 232V / 236H245R / 248D / 252K; 98L / 103A 104I / 159D / 232V / 236H / 245R248D / 252K; 98L / 102A 103A / 104I / 159D / 212G / 232V / 236H / 245R248D / 252K; 101G / 103A / 104I / I59D / 232V / 236H / 245R / 248D / 252K; 102A 103A / 104. / 159D / 232V / 236H245R / 248D / 252K; 103 A / 1041 / 159D / 232 V / 236H / 245 R / 248D / 252K; 103A 104I / 159D / 213R232V / 236H / 245R / 248D / 252; 103A / 104I / 130G / 159D / 232V / 236H / 245R / 248D / 252K; 103A 1041 / 159D / 217E / 232V / 236H / 245R / 248D / 252K; 103 A / 1041 / 159D / 248D / 252K / 270V; 103A 1041 / 159D / 232V / 236H / 245R; 103A 104I / 159D / 205I / 209W / 232V / 236H / 245R; 103AJ104I / I59D / 232V / 236H / 245R257V; 103A 104I / 159D / 205I / 209W / 232V / 236H / 245R / 257V; 103A / 1O4I / 131V / 159D / 232V / 236H / 245R / 248D / 252; 103A 104I / 159D / 205I / 209W / 210I / 232V / 236H / 245R / 257V; and 103 A / l 041 / 159D / 232V / 245RJ248D / 252K.

Recombinant Proteases / Recombinant Subtilisins A "recombinant protease" or "recombinant subtilisin" refers to a protease or subtilisin in which the DNA sequence encoding the protease or subtilisin that is presented in natural form is modified to produce a sequence of Mutant DNA encoding the substitution, insertion or deletion of one or more amino acids in the amino acid sequence of the protease or subtilisin. Suitable methods of modification are described herein, and in the US patents. Nos. RE 34,606, 5,204,015 and 5,185,258.

Proteases of non-human origin / Subtilisins of non-human origin "Proteases of non-human origin" or "subtilisins of non-human origin" and the DNA encoding them can be obtained from various prokaryotic and eukaryotic organisms. Suitable examples of prokaryotic organisms include gram negative organisms such as E. coli or Pseudomonas and gram positive bacteria such as Micrococcus or Bacillus. Examples of eukaryotic organisms from which carbonyl hydrolase and its genes can be obtained include yeasts such as the fungus Saccharomyces cerevisiae, such as the Aspergillus species and sources of non-human mammals such as, for example, the bovine species, from the which one can obtain the gene that encodes the protease chymosin or subtilisin chymosin. A series of proteases and / or subtilisins can be obtained from several related species that have amino acid sequences which are not completely homologous between the elements of said series but which nonetheless exhibit the same type or similar type of biological activity. Therefore, the non-human protease or non-human subtilisin as used herein has a functional definition that refers to proteases or subtilisins, respectively, which are associated, directly or indirectly, with prokaryotic and eukaryotic sources.

DNA Variant Sequences The sequences of the DNA variant encoding said protease or variant of subtilisin are derived from a DNA precursor sequence that encodes a recombinant or naturally occurring precursor enzyme. Sequences of the DNA variant are derived by modifying the precursor DNA sequence to encode the substitution of one or more specific amino acid residues encoded by the precursor DNA sequence corresponding to positions 103 in combination with one or more of the following positions., 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58 , 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 1 1 1, 14, 1 16, 1 17, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173 , 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 247, 249, 251, 252, 253, 254, 255, 256, 258, 259, 260, 261, 262, 263, 265, 268, 270, 271, 272, 274 and 275 of Bacillus amyloliquefaciens subtilisin; wherein, when said protease variant includes a substitution of amino acid residues at positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217 , 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin. Although the residues s of amino acids identified for modification herein are identified according to the numbering applicable to B. amyloliquefaciens (which has been the conventional method for identifying residue positions in all subtilisins), the preferred DNA precursor sequence useful for the present invention is the DNA sequence of Bacillus lentus as shown in Figure 3. In a preferred embodiment, these sequences of the DNA variant encode the substitution, insertion or deletion of the amino acid residue corresponding to position 103 of subtilisin Bacillus amyloliquefaciens in combination with one or more additional amino acid residues at positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33 , 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102 , 104, 106, 107, 109, 1 1 1, 1 14, 1 16, 1 17, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244 , 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of Bacillus amyloliquefaciens subtilisin; wherein, when each protease variant includes a substitution of amino acid residues at positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions at one or more positions of amino acid residue different from amino acid residue positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of subtilisin Bacillus amyloliquefaciens. Most preferably, this sequence of the DNA variant codes for the protease variants described herein. In another preferred embodiment, these sequences of the DNA variant encode the substitution, insertion or deletion of one or more amino acid residue corresponding to positions 62, 212, 230, 232, 252 and 257 of subtilisin Bacillus amyloliquefaciens. Most preferably, these DNA variant sequences encode the protease variants described herein. Although the amino acid residues identified for modification herein are identified according to the numbering applicable to B. amyloliquefaciens (which has been the conventional method for identifying residue positions in all subtilisins), the DNA precursor sequences useful for The present invention is the DNA sequence of Bacillus lentus as shown in Figure 3.

These recombinant DNA sequences encode protease variants having a novel amino acid sequence and, in general, at least one property that is substantially different from the same property of the enzyme encoded by the precursor protease DNA sequence. Such properties include proteolytic activity, substrate specificity, stability, altered pH profile and / or increased performance characteristics. The specific substitutions corresponding to positions 103 in combination with one to more of the following positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 1 1 1, 1 14, 1 16, 1 17, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198 203, 204, 205, 206, 209 , 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of subtilisin from Bacillus amyloliquefaciens; wherein, when each protease variant includes a substitution of amino acid residue at the positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the positions of amino acid residue corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 where the corresponding numbered positions to the subtilisin that occurs naturally from Bacillus amyloliquefaciens or to the residues of equivalent amino acids in other hydrolases carbonylhydrolases or subtilisins (such as subtilisin Bacillus lentus) are described herein. In addition, the specific substitutions corresponding to one or more of the following positions in which the numbered positions correspond to the subtilisin that occurs naturally from 62, 212, 230, 232, 252 and 257 or to the residues s of equivalent amino acids in other carbonylhydrolases or subtilisins (such as subtilisin Bacillus lentus) that are described herein. These amino acid position numbers refer to those assigned to the mature Bacillus amyloliquefaciens subtilisin sequence presented in Figure 1. The present invention, however, is not limited to the use of the mutation of its particular subtilisin but extends to the proteases precursors containing amino acid residues at positions that are "equivalent" to the particular residues s identified in Bacillus amyloliquefaciens subtilisin. In a preferred embodiment of the present invention, the precursor protease is Bacillus lentus subtilisin and the substitutions, deletions or insertions are carried out at the equivalent amino acid residue in Bacillus lentus corresponding to those listed above. A (amino acid) residue of a precursor protease is equivalent to a subtilisin residue Bacillus amyloliquefaciens if it can be homologous (ie corresponding in position in the structure either primary or tertiary) or analogous to a specific residue or portion thereof. residue in Bacillus amyloliquefaciens subtilisin (ie, having the same or similar functional ability to combine, react or interact chemically). To establish homology to the primary structure, the amino acid sequence of a precursor protease is directly compared to the primary sequence of subtilisin Bacillus amyloliquefaciens and particularly to a group of residues known as invariants in subtilisins for which said sequence is known. For example, Figure 2 in the present shows the residues conserved between the subtilisin Bacillus amyloliquefaciens and the subtilisin B. lentus. After aligning the conserved residues, the necessary insertions and deletions are allowed to maintain the alignment (ie the elimination of conserved residues is eliminated by means of the deletion and arbitrary insertion), the residues are defined equivalent to the particular amino acids in the sequence subtilisin primary Bacillus amyloliquefaciens. The alignment of conserved residues should preferably conserve 100% of said residues. However, alignment greater than 75% or less than 50% of conserved residues is also adequate to define equivalent residues. The preservation of the trivalent catalytic bond, Asp32 / His64 / Ser221, must be maintained.

For example, in Figure 3, the amino acid sequence of subtilisin from Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis (carlsbergensis) and Bacillus lentus are aligned to provide the maximum amount of homology between the amino acid sequences. A comparison of these sequences shows that there is a number of conserved residues contained in each sequence. These conserved residues (as in the case between BPN 'and Bacillus lentus) are identified in Figure 2. These conserved residues, therefore, can be used to define equivalent, corresponding amino acid residues of Bacillus lentus (Publication No. WO98). / 06279 published July 13, 1989), the preferred protease precursor enzyme herein, or subtilisin which is referred to as PB92 (EP 0 328 299), which is highly homologous to the preferred Bacillus lentus subtilisin. The amino acid sequences of certain subtiiisins of this type are aligned in Figures 3A and 3B with the subtilisin sequence Bacillus amyloliquefaciens to produce the maximum homology of conserved residues. As can be seen, there is a deletion number in the sequence of Bacillus lentus compared to the subtilisin Bacillus amyloliquefaciens. Thus, for example, the amino acid equivalent for Va1 165 in subtilisin Bacillus amyloliquefaciens in the other subtilisins is isoleucine for B. lentus and B. licheniformis. Therefore, for example, the amino acid at position +76 is asparagine (N) in both subtilisins, B. amyloliquefaciens and ß. lentus In the protease variants of the invention, however, the amino acid equivalent to the number +76 in subtilisin Bacillus amyloliquefaciens is substituted with aspartate (D). The abbreviations and one letter codes for all amino acids in the present invention are adapted to the user manual Patentin User Manual (GenBank, Mountain View, CA) 1990, p.101. "Equivalent residues" can also be defined by determining the homology at the tertiary structure level for a precursor protease whose tertiary structure has been determined by x-ray crystallography. Equivalent residues are defined as those for which the atomic coordinates of two or more of the principal chains of atoms of a particular amino acid residue of the precursor protease and of the subtilisin Bacillus amyloliquefaciens (N in N, CA in CA, C in C and O in O) is within 0.13nm and preferably 0.1nm after alignment. The alignment is achieved after the best model has been oriented and placed to give the maximum overlap of atomic coordinates of unhydrogenated protein atoms of the protease in question, to the subtilisin Bacillus amyloliquefaciens. The best model is the crystallographic model that gives the lowest R factor for the experimental defraction data at the highest available resolution. R factor = _ __ __ _ The equivalent residues that are functionally analogous to a specific residue of subtilisin Bacillus amyloliquefaciens are defined as those amino acids of the parent protease that can adopt a conformation such that it alters, modifies or contributes to the structure of ia protein, to the binding of substrate catalysis in a defined manner and attributed to a specific residue of the subtilisin Bacillus amyloliquefaciens Furthermore, they are those residues of the parent protease (for which a tertiary structure has been obtained by x-ray crystallography (which occupy a position analogous to the extent that, although the principal chain atoms of the given residue may not meet the equivalence criteria based on the occupation of a homologous position, the atomic coordinates of at least two up to the side chain atoms of the residue remain between 0.13nm of the side chain atoms of Bacillu subtilisin s amyloliquefaciens. The coordinates of the three-dimensional structure of subtilisin Bacillus amyloliquefaciens are set forth in EPO Publication No. 0 251 446 (equivalent to the patent E.U., 182,204, description of which is incorporated herein by reference) and can be used as described above, to determine residuals equivalent to the level of tertiary structure. Some of the residues identified for substitution, insertion or deletion are conserved residues while others are not. In the case that residues s that are not conserved, the replacement of one or more amino acids is limited to substitutions that produce a variant having an amino acid sequence that does not correspond to one found in nature. In the case of conserved residues, such replacements should not result in a naturally occurring sequence. The protease variants of the present invention include the mature forms of protease variants, as well as the pro- and pre-forms of said protease variants. The prepro-forms are the preferred structures since this facilitates the expression, secretion and maturation of the protease variants. "Prosequence" refers to an amino acid sequence linked to the N-terminal portion of the mature form of a protease itself that when removed results in the appearance of the "mature" form of the protease. Many proteolytic enzymes are found in nature as translational proenzyme products and, in the absence of post-translational processing, are expressed in this manner. A preferred prosequence for producing protease variants is the putative prosequence of Bacillus amyloliquefaciens subtilisin, although other protease prosequences can be used. A "signal sequence" or "pre-sequence" refers to any amino acid sequence linked to the N 'terminal portion of a protease or to the N-terminal portion of a proprotease that may participate in the secretion of mature or pro-forma forms of the protease. This definition of signal sequence is a functional definition, which means that it includes all those amino acid sequences encoded by the N-terminal portion of the protease gene that participates in the development of protease secretion under native conditions. The present invention uses said sequences to perform the secretion of protease variants as defined herein. A possible signal sequence comprises the first seven amino acid residues of the signal sequence from subtilisin Bacillus subtilis fused to the remainder of the signal sequence from Bacillus lentus_ subtilisin (ATCC 21536). A "prepro" form of a protease variant consists of the mature form of the protease having a prosequence operably linked to the amino termini of the protease and a "pre" sequence or "signal" sequence operably linked to the amino termini of the protease. the prosecution. "Expression vector" refers to a DNA construct that contains a DNA sequence that is operably linked to a suitable control sequence capable of effecting the expression of said DNA in a suitable host. Said control sequences include a promoter to effect transcription, an optional operator sequence for controlling said transcription, a sequence encoding suitable ribosome mRNA binding sites, and sequences that control the termination of transcription and translation. The vector can be a plasmid, a phage particle, or simply a potential genomic insert. Once transformed into a suitable host, the vector can replicate and function independently or the host genome, or it can, in some cases, be integrated into the genome itself. In the present description, "plasmid" and "vector" is sometimes used interchangeably as the plasmid which is the most commonly used of a vector in the present invention. However, the intention of the invention is to include such other forms of expression vectors that are useful as equivalent functions and that are, or may be known in the art. The "host cells" used in the present invention, are generally prokaryotic or eukaryotic hosts that have preferably been manipulated by the methods described in the US patent. RE 34,606 to develop them as incapable of secreting enzymatically active endoproteases. A preferred host cell for expressing protease is Bacillus strain BG2036 which is deficient in enzymatically-active neutral protease and alkaline protease (subtilisin). The construction of strain BG2036 is described in detail in the US patent. 5,264,366. Other host cells for expressing protease include Bacillus subtilis 168 (which was also described in the European patent RE 34,606 and EU patent 5,264,366, description of which is incorporated herein by reference), as well as any suitable Bacillus strain such as B. licheniformis , B. lentus, etc.). The host cells are transformed or transfected with vectors constructed using recombinant DNA techniques. Said host cells transformed with capable of either replicating vectors encoding the protease variants or expressing the desired protease variant. In the case of vectors encoding the pre-or prepro form of the protease variant, such variants, when expressed, are typically secreted from the host cell in the host cell medium. "Operably linked", when describing the relationships between two DNA regions, simply means that they are functionally related to one another. For example, a prosequence is operably linked to a peptide if it functions as a signal sequence, which participates in the secretion of the mature form of the protein, which most likely involves the cut-off site of the signal sequence. A promoter is operably linked to a coding sequence if it controls the transcription of the sequence; A ribosome binding site is operably linked to a coding sequence if it is positioned to allow translation. The genes encoding the naturally occurring precursor protease can be obtained according to the general methods known to those skilled in the art. The methods generally comprise synthesizing labeled test tubes that have putative sequences encoding regions of the protease of interest, preparing genomic libraries from organisms that express the protease, and identifying the libraries for the gene of interest by hybridizing to the test. The positively hybridizing clones are mapped and sequenced. The cloned protease is used to transform a host cell to express the protease. The protease gene is then ligated into a plasmid with a higher copy number. This plasmid replicates in hosts in the sense that it contains the well-known elements necessary for the replication of plasmids: a promoter operably linked to the gene in question (which can be supplied as the homologous promoter of the gene itself if it is recognized, ie transcribed by host), a transcription termination and a polyadenylation region (necessary for the stability of mRNA transcribed by the host from the protease gene in certain eukaryotic host cells) that is exogenous or is supplied by the endogenous terminator region of the host gene. protease and, desirably, a selection gene such as an antibiotic resistance gene that allows maintenance of continuous culture of host cells infected with plasmid by growth in a medium containing antibiotics. The plasmids with the highest copy number also contain an origin of replication for the host, thus providing large numbers of plasmids to be generated in the cytoplasm without chromosomal imitation. However, within the scope of the present, it is to integrate multiple copies of the protease gene into the host genome. This is facilitated by prokaryotic and eukaryotic organisms that are particularly susceptible to homologous recombination. The gene can be a natural S. lentus gene. Alternatively, a synthetic gene encoding a naturally occurring mutant precursor protease can be produced. In such an approach, the DNA and / or amino acid sequence of the precursor protease is determined. Subsequently, multiple DNA fragments, synthetic single chain and overlapping are synthesized, which, upon hybridization and ligation, produce a synthetic DNA that includes the precursor protease. An example of synthetic gene construction is set forth in Example 3 of the US patent. 5,204,105, description of which is incorporated herein by reference. Once the synthetic or naturally occurring precursor protease gene has been cloned, a number of modifications are carried out to increase the use of the gene beyond the synthesis of the naturally occurring precursor protease. Such modifications include the production of recombinant proteases as described in the US patent. RE 34,606 and in EPO Publication No. 0 251 446 and the production of protease variants described herein. The following method of cassette mutagenesis can be used to facilitate the construction of the protease variants of the present invention, although other methods can also be used. First, the naturally occurring gene encoding the protease is obtained and sequenced in whole or in part. The sequence is then identified for a point at which a mutation is desired (deletion, insertion or substitution, of one or more amino acids in the encoded enzyme.) The sequences that flank this point are evaluated for the presence of restriction sites to replace a short segment of the gene with a group or well of oligonucleotide whichWhen it is expressed it will code several mutants. Said restriction sites are preferably unique sites within the protease gene to facilitate replacement of the segment of the gene. However, any restriction or convenient restriction site that is not redundant in excess in the protease gene can be used, since the gene fragments generated by restriction digestion can be reassembled in the proper sequence. If the restriction sites are not present in locations within a convenient distance from the selected point (from 10 to 15 nucleotides) said sites are generated by substituting nucleotides in the gene in such a way that neither the reading frame nor the amino acids coded are changed in the final construction. The mutation of the gene is achieved, so that it can change its sequence and conform to the desired sequence by the first extension M13, according to the generally known methods. The task of locating the appropriate flanking regions and evaluating the changes necessary to arrive at the two convenient restriction site sequences are routinely carried out by the redundancy of a genetic code, a restriction enzyme map of the gene and the large number of different restriction enzymes. Note that if a suitable flanking restriction site is available, the above method needs to be used only in the flanking region that does not contain the site. Once the naturally occurring DNA or the synthetic DNA is cloned, the restriction sites flanking the positions to be mutated are digested with the bound restriction enzymes and / or a plurality of terminal oligonucleotide cassettes. Complementary endpoints are linked in the gene. Mutagenesis is simplified by this method because all oligonucleotides can be synthesized to have the same restriction sites, and synthetic linkers are not necessary to create said restriction sites. As used herein, proteolytic activity is defined as the rate of hydrolysis of peptide bonds per milligram of active enzyme. There are many well known methods for measuring proteolytic activity (K.M. Kalisz, "Microbial Proteinases", Advances in Biochemical Enqineerinq / Biotechnoloqy, A. Fiechter ed., 1988). ("Microbial Proteinases" Advances in Biochemical Engineering / Biotechnology) In addition to, or as an alternative for modified proteolytic activity, the variant enzymes of the present invention may have other modified properties such as the ratio Km, kcat, kcat Km and / or the specifically modified substrate and / or the modified pH activity profile. These enzymes can be custom-made for the particular substrate itself which is considered in advance as present, for example in the preparation of peptides or for hydrolytic processes such as for laundry uses. In one aspect of the invention, the aim is to ensure a variant protease that has altered proteolytic activity compared to the precursor protease, since by increasing said activity (numerically greater) it allows the use of the enzyme to act more efficiently on a target substrate. Also of interest are variant enzymes that have altered thermal stability and / or specific character of altered substrate compared to the precursor. However, in some cases a lower proteolytic activity may be desirable, for example a reduction in proteolytic activity would be useful where the synthetic activity of the proteases is desired (to synthesize peptides). It would be desirable to reduce this proteolytic activity, which is capable of destroying the product of said synthesis. On the contrary, in some cases, it may be desirable to increase the proteolytic activity of the variant enzyme against its precursor. Additionally, the increases or reductions (alteration) of the stability of the variant, either alkaline or thermal stability, may be desirable. The increments or decrements in kcat, Km or K ^ / Km are specific for the substrate used to determine these kinetic parameters. In another aspect of the invention, it has been determined that substitutions at positions corresponding to 103 in combination with one or more of the following positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18 , 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86 , 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 1 1 1, 1 14, 1 16, 117, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240 , 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270 , 271, 272, 274 and 275 of subtilisin Bacillus amyloliquefaciens are important for modulating the stability in general and / or the proteolytic activity of the enzyme. In a further aspect of the invention, it has been determined that substitutions in one or more of the following positions correspond to positions 62, 212, 230, 232, 252 and 257 of Bacillus amyloliquefaciens subtilisin which are also important for modulating stability in general and / or the proteolytic activity of the enzyme. These substitutions are preferably carried out in Bacillus lentus (recombinant or native type), although substitutions can be made in any Bacillus protease. Based on the selection results obtained with the variant proteases, the mutations identified in Bacillus amyloliquefaciens subtilisin are important for the proteolytic activity, performance performance and / or stability of these enzymes and the cleaning or washing performance of said variant enzymes. The methods and methods for making the enzymes used in the detergent and cleaning compositions of the present invention are well known and are described in PCT Publication No. WO95 / 10615. The enzymes of the present invention have specific character of trypsin forms. That is, the enzymes of the present invention hydrolyze proteins by preferentially cleaving the peptide bonds of charged amino acid residues, very specifically residues such as arginine and lysine, instead of preferentially cleaving the peptide bonds of hydrophobic amino acid residues, very specifically phenylalanine, tryptophan and tyrosine. The enzymes that have this last profile have a specific character in the form of chymotrypsin. As mentioned above, the specific character of the substrate is illustrated by the action of the enzyme on two synthetic substrates. The protease having a specific character in the form of trypsin hydrolyses the synthetic substrate bVGR-pNA preferably on the synthetic substrate sucAAPF-pNA. In contrast, protease enzymes in the form of chymotrypsin, hydrolyze the latter much faster than the previous one. For the purposes of the present invention, the following procedure was employed to define the specific character in the form of trypsin of the protease enzymes of the present invention: A fixed amount of a glycine pH regulator is added at a pH of 10 a temperature of 0.5 ppm to a standard 10 ml test tube. 1.25 mg of the active enzyme that is going to be tested in the test tube is added. Approximately, 1.25 mg of synthetic substrate per ml of regulated solution is added to the test tube. The mixture is allowed to incubate for 15 minutes at 25 ° C. At the end of the incubation period, an enzyme inhibitor, PMSF, is added to the mixture at a level of 0.5 mg per ml of pH-regulated solution. The absorbance or OD value of the mixture is read at 410 nm wavelength. Absorbency then indicates the activity of the enzyme in the synthetic substrate. The greater the absorbency, the higher the level of activity against said substrate. Then to determine the specific character of an individual enzyme, the absorbance in the two synthetic substrate proteins can be converted into a specific character relationship. For the purposes of the present invention, the ratio is determined by the specific character of the formula of: [activity in sAAPF-pNA] / [activity in bVGR-pNA] An enzyme having a ratio of less than about 10, most preferably less about 5 and most preferably about 2.5 can be considered to demonstrate activity in the form of trypsin. Said variants generally have at least one property that is different from the same property of the protease precursor from which the amino acid sequence of the variant is derived. One aspect of the composition is compositions, such as detergents and cleaning agents for the treatment of textiles, tableware, tableware, kitchenware, kitchenware, and other hard surface substrates including one or more variant proteases of the present invention. invention. The compositions containing proteases can be used to treat, for example: silk or wool, as well as other types of fabrics, as described in publications such as RD 216,034, EP 134,267, US 4,533,359 and EP 344,259; and dinnerware, tableware, kitchenware, kitchenware and other hard surface substrates as described in publications such as E.U.A. 5,478,742, E.U.A. 5,346,822, E.U.A. 5,679,630, and E.U.A. 5,677,272.

Cleaning compositions The cleaning compositions of the present invention also comprise, in addition to one or more protease variants described hereinbefore, one or more cleaning adjunct materials, preferably compatible with the protease variants. The term "cleaning attachment materials" as used herein, means any liquid, solid or gaseous selected from the particular type of cleaning composition desired and the product form (e.g., liquid, granular, powder; in paste, in aerosol, in tablet, in gel, in foam) whose materials are also preferably compatible with the protease enzyme used in the composition. The granulated compositions can also be in "compact" form and the liquid compositions can also be in "concentrated" form. The specific selection of cleaning attachment materials can easily be made by considering the surface, article or fabric to be cleaned, and the desired form of the composition for cleaning conditions during use (e.g., by the use of household detergent). washed). The term "compatible", as used herein, means cleaning composition materials that do not reduce the proteolytic activity of the protease enzyme to such an extent that the protease is not effective as desired during situations of normal use. Examples of suitable cleaning adjunct materials include, but are not limited to, surfactants, builders, bleaches, bleach activators, bleach catalysts, other enzymes, enzyme stabilizer systems, chelators, optical brighteners, soil release polymers. dye transfer agents, dispersants, foam suppressors, dyes, perfumes, dyes, fillers or fillers, hydrotropes, photoactivators, fluorescence activators, fabric conditioners, hydrolyzable surfactants, preservatives, antioxidants, anti-caking agents, anti-wrinkle agents, germicides, fungicides, colored specks, silver care agents, anti-rust agents and / or anticorrosive agents, alkalinity sources, solubilizing agents, vehicles, processing aids, pigments and pH control agents as described in the patents USA Nos. 5,705,464, 5,710.1 15, 5,698,504, 5,695,679, 5,686,014 and 5,646,101. The materials for the specific cleaning composition are exemplified in detail herein below. If the cleaning adjunct materials are not compatible with the protease variants in the cleaning compositions, then suitable methods can be used to keep the cleaning adjunct materials and the protease variants separated (not in contact with each other or with each other) until the combination of the two components is appropriate. Suitable methods can be any method known in the art, such as gel capsule, encapsulation, tablets, physical separation, etc. Preferably, an effective amount of one or more protease variants described above is included in the compositions useful for cleaning a variety of surfaces that require the removal of proteinaceous spots. Said cleaning compositions include detergent compositions for cleaning hard surfaces, unlimited in shape (for example liquid and granular); detergent compositions for fabric cleaning, unlimited in shape (for example, granular, liquid and stick formulations); dishwashing compositions (unlimited in shape and including both liquid and granular forms of automatic dishwashing); compositions for oral cleansing, limited in form (for example, toothpaste, toothpaste and mouthwash formulations); compositions for denture cleaning, unlimited in shape (for example, liquid, in tablets). As used herein, "effective amount of a protease variant" refers to the amount of protease variant described herein above, necessary to achieve the enzymatic activity that is required in the specific cleaning composition. Said effective amounts can be easily achieved by the person skilled in the art and are based on various factors, such as the particular variant used, the cleaning application, the specific composition of the cleaning composition, and whether a liquid composition is required or a dry (for example, granulated, in bar), and the like. Preferably, the cleaning compositions comprise from about 0.0001% preferably about 0.001%, most preferably about 0.01% by weight of the cleaning compositions of one or more protease variants of the present invention, to about 10%, preferably about 1%, most preferably at about 0.1%. In addition, preferably the protease variant of the present invention is present in the compositions in an amount sufficient to provide a milligram ratio of active protease per 100 grams of theoretical available O2 ("AvO2") composition of any peroxyacid in the solution washing, which is referred to herein as the Enzyme to Bleach ratio (E / B ratio), which varies from about 1: 1 to about 20: 1. Various examples of various cleaning compositions may be employed wherein the protease variants of the present invention are discussed in greater detail below. In addition, the cleaning compositions may include from about 1% to about 99.9% by weight of the composition of the cleaning adjunct materials. The cleaning compositions of the present invention can be in the form of "fabric cleaning compositions" or "non-fabric cleaning compositions". As used herein, "fabric cleaning compositions" includes laundry detergent compositions either manually and by machine including active laundry compositions and compositions suitable for use in soaking and / or pretreatment of soiled fabrics. As used herein, "non-fabric cleaning compositions" includes hard surface cleaning compositions, dishwashing detergent compositions, oral cleaning compositions, denture cleaning compositions, and personal cleansing compositions. When the cleaning compositions of the present invention are formulated as compositions suitable for use in a laundry machine washing method, the compositions of the present invention preferably contain both a surfactant and a builder compound and additionally, one or more cleaning adjunct materials, preferably selected from organic polymeric compounds, bleaching agents, additional enzymes, suds suppressants, dispersants, lime soap dispersants, dirt slurry and anti-redeposition agents and corrosion inhibitors. The laundry compositions may also contain softening agents, as additional cleaning adjunct materials. The compositions of the present invention can also be used as detergent additive products in solid or liquid form. Said additive products are intended to supplement or boost the performance of conventional detergent compositions and can be added at any stage of the cleaning process. When formulated as compositions for use in manual dishwashing methods, the compositions of the invention preferably contain a surfactant and preferably other auxiliary materials selected from organic polymeric compounds, foaming agents, group II metal ions, solvents., hydrotropes and additional enzymes.

If required, the density of the laundry detergent compositions herein ranges from 400 to 1200 g / liter, preferably from 500 to 950 g / liter of composition, measured at 20 ° C. The "compact" form of the compositions of cleaning of the present invention is best reflected by the density and, in terms of composition, by the amount of inorganic filler salt; the inorganic filler salts are conventional ingredients of detergent compositions in powder form; in conventional detergent compositions, the filler salts are present in substantial amounts, typically 17-35% by weight of the total composition. In compact compositions, the filler salt is present in amounts not exceeding 15% of the total composition, preferably not exceeding 10%, most preferably not exceeding 5% by weight of the composition. The inorganic filler salts, as indicated in the present compositions, are selected from the alkali salts and alkaline earth metals of sulfates and chlorides. A preferred filler salt is sodium sulfate. The liquid cleaning compositions according to the present invention can also be in "concentrated form", in that case, the liquid cleaning compositions according to the present invention will contain a smaller amount of water, compared with conventional liquid detergents. Typically, the water content of the concentrated liquid cleaning composition is preferably less than 40%, most preferably less than 30%, most preferably still less than 20% by weight of the cleaning composition.

Cleaning Attachments Surfactant System The detersive surfactants included in the fully formulated cleaning compositions encompassing the present invention comprise at least 0.01%, preferably at least about 0.1%, most preferably at least about 0.5%, most preferably at least 1% to about 60%, most preferably to about 35% by weight of the cleaning composition depending on the particular surfactants used and the desired effects. The detersive surfactant may be nonionic, anionic, ampholytic, zwitterionic, cationic, semi-polar non-ionic and mixtures thereof, non-limiting examples, which are described in U.S. Patent Nos. 5,707,950 and 5,576,282. Preferred detergent and cleansing compositions comprise nonionic detersive surfactants or mixtures of anionic surfactants with other surfactants, especially nonionic surfactants. Non-limiting examples of surfactants useful herein include C.? 18 alkyl benzene sulphonates. and primary, secondary and random alkylsulphates, C-io-C-alkylaryl sulfates, C10-? β alkylarylglucosides and their corresponding sulfated polyglycosides, alpha-sulfonated fatty acid esters of C2-C8, alkyl of C12-C? s and alkylphenol alkoxylates (especially mixed ethoxylates and ethoxy / propoxy), C? 2-C? beta betaines and sulfobetaines, ("sultaines"), C .oC .ß amine oxides, and the like. Other conventional useful surfactants are described in standard texts. The surfactant is preferably formulated to be compatible with the enzyme components present in the composition. In liquid or gel compositions, the surfactant is most preferably formulated in a manner that promotes, or at least does not degrade, the stability of any enzyme in these compositions.

Nonionic surfactants The condensates of polyethylene oxide, polypropylene and polybutylene of alkylphenols are suitable for use as the nonionic surfactant of the surfactant systems of the present invention, with the polyethylene oxide condensates being more preferred. Commercially available nonionic surfactants of this type include Igepa.TM CO-630, marketed by GAF Corporation; and Triton ™ X-45, X-1 14, X-100 and X-102, all marketed by Rohm & Haas Company. These surfactants are commonly known as alkylphenol alkoxylates (alkylphenol ethoxylates). The condensation products of the primary and secondary aliphatic alcohols with about 1 to about 25 moles of ethylene oxide are suitable for use as the nonionic surfactant of the nonionic surfactant system of the present invention. Examples of commercially available nonionic surfactants of this type include Tergitol ™ 15-S-9 (the linear alcohol condensation product of C-j -j -C-i s with 9 moles of ethylene oxide), Tergitol ™ 24-L-6 NMW (the primary alcohol condensation product of C-J2-C-I4 with 6 moles of ethylene oxide with a limited molecular weight distribution), both marketed by Union Carbide Corporation; Neodol ™ 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 C12-C1.3 with 3.0 moles of ethylene), Neodol ™ 45-7 (the linear alcohol condensation product of C14-C15 with 7 moles of ethylene oxide), Neodol ™ 45-5 (the linear condensation product of C14-C15 with 5 moles of ethylene oxide) marketed by Shell Chemical Company, Kyro ™ EOB (the condensation product of C13-C-15 alcohol with 9 moles of ethylene oxide), marketed by The Procter & Gamble Company, and Genapol LA O3O or O5O (the condensation product of C-12-C14 alcohol with 3 or 5 moles of ethylene oxide) marketed by Hoechst. The preferred scale of HLB in these products is 8-11 and most preferred is 8-10. Also useful as the nonionic surfactant of the surfactant systems of the present invention are the alkyl polysaccharides described in the US patent. No. 4,565,647. Preferred alkyl polyglycosides have the formula R2? (CnH2nO) t (glucosyl) x wherein R2 is selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl groups and mixtures thereof, in which 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 from about 1.3 to about 3, most preferably from about 1.3 to about 2.7. The condensation products of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide with propylene glycol are also suitable for use as the additional nonionic surfactant system of the present invention. Examples of compounds of this type include certain Plurafac ™ LF 404 surfactants commercially available as Piuronic ™, marketed by BASF. Also suitable for use as the nonionic surfactant of the nonionic surfactant system of the present invention are the condensation products of ethylene oxide with the product resulting from the reaction of propylene oxide and ethylene diamine. Examples of this type of nonionic surfactant include certain of the commercially available Tetronic ™ compounds, marketed by BASF. Preferred to be used as the nonionic surfactant of the surfactant systems of the present invention are the polyethylene oxide condensates of alkylphenols, the condensation products of primary and secondary aliphatic alcohols with from about 1 to about 25 moles of ethylene, alkyl polysaccharides and mixtures thereof. The most preferred are alkylphenol ethoxylates of C8-C-J4 having from 3 to 15 ethoxy groups and alcohol ethoxylates of Cs-C-is (preferably from CJ O average) having from 2 to 10 ethoxy groups , and mixtures thereof. The highly preferred nonionic surfactants are the polyhydroxy fatty acid amide surfactants of the formula: R2-C (O) -N (R1) -Z wherein R1 is H, or R is C1-C4 hydrocarbyl, 2-hydroxyethyl, 2-hydroxypropyl or a mixture thereof, R2 is C5-31 hydrocarbyl and Z is polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with at least 3 hydroxyls directly connected to the chain, or an alkoxylated derivative thereof . Preferably, R1 is methyl, R2 is an alkyl chain of C- | 1-C-15 or straight CJ QCJ S alkyl or alkenyl such as coconut alkyl or mixtures thereof, and Z is derived from a reducing sugar such as glucose, fructose, maltose and lactose, in a reductive amination reaction .

Anionic surfactants Suitable anionic surfactants to be used are alkyl benzene sulfonate surfactants, alkyl ester sulfonate which include linear esters of C8-C20 carboxylic acids (ie, fatty acids) which are sulfonated with gaseous SO3 in accordance with "The Journal of the American Oil Chemists Society ", 52 (1975), pp. 323-329. Suitable starting materials could include natural fatty substances such as those derived from tallow, palm oil, etc. The preferred alkyl ester sulfonate surfactant, especially for laundry applications, comprises alkyl ester sulfonate surfactants of the structural formula: wherein R3 is a C8-C20 hydrocarbyl. preferably a alkyl or combination thereof, R4 is a Ci-Cg hydrocarbyl, preferably an alkyl or a combination thereof, and M is a cation which forms a water-soluble salt with the alkyl ester sulfonate. Suitable salt-forming cations include metals such as sodium, potassium and lithium, and substituted or unsubstituted ammonium cations such as monoethanolamine, diethanolamine and triethanolamine. Preferably, R3 is C-J alkyl Q-C-16 and R is methyl, ethyl or isopropyl. It preferred especially the methyl ester sulfonates wherein R3 is C- alkyl; Q- C-16 Other suitable anionic surfactants include the alkyl sulfate surfactants which are salts or water soluble acids of the formula ROSO3M, wherein R is preferably a hydrocarbyl of C-J0-C24, preferably an alkyl or hydroxyalkyl having an alkyl component of C10-C20. most preferably an alkyl or C12-C18 hydroxyalkyl. and M is H or a cation. Typically, C12-16 alkyl chains are preferred for lower wash temperatures (e.g., below about 50 ° C) and alkyl chains of Cj g-> s are preferred for higher wash temperatures (e.g. about 50 ° C). Other anionic surfactants useful for the detersive purposes include soap salts, C8-C22 primary or secondary alkane sulphonates. C8-C24 olefinsulfonates, sulfonated polycarboxylic acids prepared by the sulfonation of the pyrolyzed product of alkali earth metal citrates, for example, as indicated in the description of British Patent No. 1, 082,179, C8-C24 alkyl polyglycol ether sulfates (containing up to 10 moles of ethylene oxide); alkyl glycerol sulfonates, fatty acyl glycerol sulfonates, oleylglycerol fatty sulphonates, ethylene oxide sulphates of alkylphenol, parafin sulfonates, alkyl phosphates, isethionates, such as acyl isethionates, N-acyl taurates, alkyl succinamates and sulfosuccinates, monoesters of sulfosuccinates (especially saturated and unsaturated C 4 -Ci-saturated monoesters) and sulfosuccinate diesters (especially saturated and unsaturated CQ-C-2 diesters), acyl sarcosinates, alkylpolyacharide sulfates such as alkyl polyglucoside sulfates (the non-sulphonated non-ionic compounds which described below), branched primary alkyl sulphates and alkyl polyethoxycarboxylates such as those of the formula RO (CH 2 CH 2?) [-CH 2 COO-M + wherein R is a C 8 -C 22 alkyl. k is an integer from 1 to 10 and M is a soluble salt forming cation. Resin acids and hydrogenated resin acids are also suitable, such as rosin, hydrogenated rosin and resin acids, as well as hydrogenated resin acids present in or derived from tallow oil. Additional examples are described in "Surface Active Agents and Detergents" (Vol. I and II by Schwartz, Perry and Berch). A variety of such surfactants are also generally described in the US patent. No. 3,929,678, issued December 30, 1975 to Laughiin, et al, in Column 23, line 58 to Column 29, line 23 (incorporated herein by reference). The most preferred anionic surfactants include the alkoxylated alkylsulfate surfactants which are water soluble salts or acids of the formula RO (A) mSO3M wherein R is an alkyl or hydroxyalkyl group of unsubstituted Cj n-C24 having an alkyl component of CI Q-C24, preferably an alkyl or hydroxyalkyl of Cl 2"C-20", most preferably C 1-12 alkyl or hydroxyalkyl, A is an ethoxy or propoxy unit, m is greater than zero, typically between about 0.5 and about 6, most preferably between about 0.5 and about 3 and M is H or a cation which may be, for example, a metal cation (for example, sodium, potassium, lithium, calcium, magnesium, etc.) or an ammonium or substituted ammonium cation. present the ethoxylated alkyl sulphates as well as the propoxylated alkyl sulphates Specific examples of substituted ammonium cations include methyl-, dimethyl- and trimethyl ammonium cations and quaternary ammonium cations such as tetramethylammonium and dimethylpiperidinium cations and those derived from aquilamines such as ethylamine, diethylamine, triethylamine, mixtures thereof and the like Exemplary surfactants are polyethoxylated alkyl sulphate of C12-C-18 (1 -0) (Ci2-Ci 8E (1 -0) M), alkyl sulfate polyethoxylate of C12-C18 (2-25) (C12-Ci 8 - = (2.25) M), polyethoxylated alkyl sulfate of C12-CI 8 (-0) (C2-CJ 8E (3.0) M), and alkyl sulfate polyethoxylated of C12-C18 (4- °) C- | 2-C. dE (4.0) M), in which M is conveniently selected from sodium and potassium. When included therein, the detergent compositions of the present invention typically comprise about 1%, preferably about 3% to about 40%, preferably about 20%, by weight of said anionic surfactants.

Cationic Surfactants The cationic detersive surfactants suitable for use in the detergent compositions of the present invention are those having a long chain hydrocarbyl group. Examples of such cationic surfactants include ammonium surfactants such as alkyltrimethyl ammonium halides and those surfactants having the formula: [R2 (OR3) y] [R4 (OR3) y_2R5N + X- where 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 consisting of -CH2CH2-, -CH2CH (CH3) -, - CH2CH (CH2? H) -, -CH2CH2CH2-, and mixtures thereof; each R4 is selected from the group consisting of C-1-C4 alkyl, hydroxyalkyl of C-J-C4, benzyl ring structures formed by joining the two groups R4, -CH2CHOH-, -CHOHCOR6CHOHCH2OH, wherein R6 is any hexose or hexose polymer having a molecular weight less than about 1000, and hydrogen when and not being 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 y is from 0 to approximately 15; and X is any compatible anion.

The most preferred cationic surfactants are the water-soluble quaternary ammonium compounds useful in the present composition, having the formula (1): R 2 R 3 R 4 N + X ~ wherein R 1 is C 1 -C 4 alkyl, each of R 2 , R3 and R4 is independently alkyl of C1-C4, hydroxyalkyl of C-J-C4, benzyl and - (C2H4o)? '~', wherein x has a value of 2 to 5 and X is an anion. No more than one of R2, R3 or R4 must be benzyl. The preferred length of the alkyl chain for Rj is C-12-C15, particularly when the alkyl group is a mixture of chain lengths derived from palm kernel or coconut oil or is synthetically derived by the olefin accumulation or the synthesis of OXO alcohols. Preferred groups for R, R3 and R4 are methyl and hydroxyethyl groups, and the anion X can be selected from halide, methosulfate, acetate and phosphate ions. Examples of quaternary ammonium compounds of the formula (i) for use herein include, but are not limited to: coconut trimethyl ammonium chloride or bromide; coconut methyl dihydroxyethyl ammonium chloride or bromide; decyl triethyl ammonium chloride; decyl dimethyl hydroxyethyl ammonium chloride or bromide; C12-C1.5 dimethyl hydroxyethyl ammonium chloride or bromide; coconut dimethyl hydroxyethyl ammonium chloride or bromide; Methyl Trimethyl Ammonium Methyl Sulfate; lauryl dimethyl benzyl ammonium chloride or bromide; lauryl dimethyl (ethenoxy) 4 ammonium chloride or bromide; choline esters (compounds of the formula i) in which R- | is alkyl of CH2-CH2-O-C-C-12-14 and 2R3R4 are O-methyl) and di-alkyl imidazolines [(i)]. Other cationic surfactants useful herein are also described in the U.S. patent. No. 4,228, 044, Cambre, issued October 14, 1980, and in the European patent application EP 000,224. When included therein, the detergent compositions of the present invention typically comprise about 0.2%, preferably about 1% to about 25%, preferably about 8% by weight of said cationic surfactants.

Ampholytic Surfactants Ampholytic surfactants are also suitable for use in the cleaning compositions of the present invention, examples of which are described in the U.S. patent. No. 3,929,678. When included therein, the cleaning compositions of the present invention typically comprise about 0.2%, preferably about 1% to about 15%, preferably about 10%, by weight of said ampholytic surfactants.

Zwitterionic surfactants Zwitterionic surfactants, examples of which are described in US Pat. No. 3,929,678, are also suitable for use in cleaning compositions.

When included therein, the detergent compositions of the present invention typically comprise 0.2%, preferably from about 1% to about 15%, preferably to about 10% by weight of said zwitterionic surfactants.

Semi-polar nonionic surfactants Semi-polar nonionic surfactants are a special category of nonionic surfactants including water-soluble amine oxides having the formula: 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 can be attached one to the other). another, e.g., through an oxygen or nitrogen atom to form a ring structure); water-soluble phosphine oxides containing an alkyl portion of from about 10 to about 18 carbon atoms and a portion selected from the group consisting of alkyl and hydroxyalkyl portions of from about 1 to about 3 carbon atoms. Amine oxide surfactants include, in particular, alkyl dimethylamine oxides of C ^ \ Q-C ^ Q and alkoxyethyldihydroxyethylamine oxides of C8-C < 2- When included therein, the cleaning compositions of the present invention typically comprise from about 0.2%, preferably from about 1% to about 15%, preferably to about 10% by weight of said semi-polar non-ionic surfactants .

Co-surfactants The cleaning compositions of the present invention may further comprise a co-surfactant selected from the group of primary or tertiary amines. Primary amines suitable for use herein include amines according to the formula R-j NH 2, wherein R 1 is an alkyl chain of C 2 -C 2+; preferably Cg-C-j or. or R4X (CH2) n. X is -O-, - C (O) NH- or -NH-, R4 is an alkyl chain of C5-C-12, n is between 1 to 5, preferably 3. The alkyl chains of Rj can be straight or branched and they may be interrupted with up to 12, preferably less than 5, portions of ethylene oxide. Preferred amines according to the above formula are the n-alkylamines. Amines suitable for use herein may be selected from 1-hexylamine, 1-octylamine, 1-decylamine and laurylamine. Other preferred primary amines include Cs-C ^ o oxypropylamine. octyloxypropylamine, 2-ethylexyl-oxypropylamine, lauryl amido propylamine and amido propylamine. Most preferred amines for use in the compositions of the present invention include 1-hexylamine, 1-octylamine, 1-decylamine, 1-dodecylamine. Especially desirable are n-dodecyldimethylamine and bishydroxyethylcocoalkylamine and 7-fold ethoxylated oleylamine, lauryl amido propylamine and cocoamidopropylamine.

LFNI (Non-ionic surfactants with low foaming effect) The particularly preferred surfactants in the compositions of the present invention for automatic dishwashing (ADD) are the low foaming nonionic surfactants (LFNI) which are described in EU patents Nos. 5,705, 464 and 5,710,115. The LFNI can be present in amounts of 0.01% to about 10% by weight, preferably from about 0.1% to about 10%, and most preferably from about 0.25% to about 4%. The LFNI are used very typically in automatic dishwashing (ADD) taking into account its improved water-rolling action (especially glass, which confers to the dishwashing product (ADD).) Also include materials without silicone , polymeric without phosphate, which are illustrated hereinafter, which are known to release the foaming of the food residues found in automatic dishwashing, LFNI include non-ionic alkoxylated surfactants, especially ethoxylates derived from primary alcohols, and mixtures thereof with more sophisticated surfactants, such as polyoxypropylene / polyoxyethylene / polyoxypropylene (PO / EO / PO) reverse block polymers as described in US Pat. Nos. 5,705,464 and 5,710,115. can also be used include those non-ionic surfactants POLY-TERGENT® SLF-18 from Olin Corp., and any Biodegradable LFNI having the melting point properties, described above. These and other nonionic surfactants are well known in the art, which are described in more detail in Kirk Othmer's Encyclopedia of Chemical Technology, 3a. Edition, Vol. 22, pp 360- 379, "Surfactants and Detersive Systems", incorporated herein by reference.

Bleaching System The compositions of the present invention preferably contain a bleaching system. Bleaching systems typically include a "bleaching agent" (source of hydrogen peroxide) and a "initiator" or "catalyst". When present, bleaching agents will typically be present at levels of about 1%, preferably from about 5% to about 30%, preferably about 20%, by weight of the composition. If present, the amount of bleach activator will typically be about 0.1%, preferably about 0.5% to about 60%, preferably about 40%, by weight, of the bleaching composition comprising the bleaching agent-plus -activator bleach.

Blanching agents Sources of hydrogen peroxide are described in detail in the reference incorporated herein, Kirk Othmer's Encyclopedia of Chemical Technology, 4a. Edition, (1992, John Wiley &Sons), Vol. 4, pp. 271 -300"Bleaching Agents (Survey)", and includes the various forms of sodium perborate and sodium percarbonate, including various coated and modified forms. The preferred source of hydrogen peroxide which is used herein may be any convenient source, including the hydrogen peroxide itself. For example, perborate, e.g., sodium perborate (any hydrate but preferably the mono- or tetrahydrate), sodium carbonate peroxyhydrate or equivalent percarbonate salts, sodium pyrophosphate peroxyhydrate, urea peroxyhydrate or peroxide sodium, can be used in the present. Also available are available oxygen sources such as persulfate bleach (e.g., OXONE, manufactured by DuPont). Sodium perborate monohydrate and sodium percarbonate are particularly preferred. Any mixture of any convenient source of hydrogen peroxide can also be used. A preferred percarbonate bleach comprises dry particles having an average particle size on the scale of about 500 microns to about 1., 000 microns, no more than about 10% by weight of said particles that are less than about 200 microns and no more than about 10% by weight of said particles that are greater than about 1.250 microns. Optionally, the percarbonate can be coated with a silicate, borate or with water soluble surfactants. Percarbonate is available from commercial sources such as FMC, Solvay and Tokai Denka. The compositions of the present invention may further comprise as the bleaching agent a chlorine bleaching material. Such agents are well known in the art, and include for example sodium dichloroisocyanurate ("NaDCC"). However, chlorine-type bleaches are the least preferred for compositions containing enzymes. (a) Bleach activators Preferably, the peroxygen bleach component in the composition is formulated with an activator (peracid peracid). The activator is present at levels of about 0.01%, preferably about 0.5%, most preferably from about 1% to about 15%, preferably about 10%, most preferably about 8%, by weight of the composition. Preferred activators are selected from the group consisting of tetraacetylethylenediamine (TAED), benzoylcaprolactam (BzCL), 4-nitrobenzoylcaprolactam, 3-chlorobenzoylcaprolactam, benzoyloxybenzenesulfonate (BOBS), nonanoyloxybenzenesulfonate (NOBS), phenylbenzoate (PhBz), decanoyloxybenzenesulfonate (do-OBS), benzoylvalerolactam (BZVL), octanoyloxybenzenesulfonate (C8-OBS), perihydrolyzable esters and mixtures thereof, most preferably benzoylcaprolactam and benzoylvalerolactam. Particularly preferred bleach activators in the pH range ranging from 8 to about 9.5 are those selected having an OBS or VL leaving group. Preferred hydrophobic bleach activators include, but are not limited to, nonaoyloxybenzenesulfonate (NOBS), sodium 4- (N-nonanoyl) amino hexanoyloxy-benzene sulfonate salt (NACA-OBS) whose example is described in US Pat. 5,523,434, dodecanoyloxybenzenesulfonate (LOBS or C.2-OBS, 10-undecenoyloxybenzenesulfonate (UDOBS or Cn.OBS, with unsaturation at position 10), and decanoyloxybenzoic acid (DOBA) Preferred bleach activators are those described in US 5,698,504 Christie et al., Issued December 16, 1997; EU 5,695,679 Christie et al., Issued December 9, 1997; ,686,401 Willey et al., Issued November 11, 1997; E.U. 5,686,014 Hartshorn et al., Issued November 1, 1997; E.U. 5,405,412 Willey et al., Issued April 1, 1995; E.U. 5,405,413 Willey et al., Issued April 1, 1995; E.U. 5,130,045 Mitchel et al., Issued July 14, 1992; and E.U. 4,412,934 Chung et al., Issued November 1, 1983, and copending patent applications serial numbers 08 / 709,072, 08 / 064,564, all of which are incorporated herein by reference. The molar ratio of the peroxygen bleach compound (as AvO) to bleach activator in the present invention, generally ranges from at least 1: 1, preferably from about 20: 1, most preferably from about 10: 1 to about 1 : 1, preferably around 3: 1. The substituted quaternary bleach activators can also be included. The present cleaning compositions preferably comprise a quaternary substituted bleach activator (QSBA) or a substituted quaternary peracid (QSP); most preferably, the previous one. The preferred QSBA structures are further described in E.U. 5,686,015 Willey et al., Issued November 1, 1997; E.U. 5,654,421 Taylor et al., Issued Aug. 5, 1997; E.U. 5,460,747 Gosselink et al., Issued October 24, 1995; E.U. 5,584,888 Miracle et al., Issued December 17, 1996; and E.U. 5,578,136 Taylor et al., Issued November 26, 1996; all of them are incorporated herein by reference.

The highly preferred bleach activators useful herein are amide-substituted as described in E.U. 5,698,504, E.U. 5,695,679 and E.U. 5,686,014 each of which was cited hereinbefore. Preferred examples of said bleach activators include: (6-octanamidocaproyl) oxybenzenesulfonate, (6-nonanamidocaproyl) oxybenzenesulfonate, (6-decanamidocaproyl) oxybenzenesulfonate and mixtures thereof. Other useful activators, described in E.U. 5,698,504, E.U. 5,695,679, and E.U. 5,686,014 each of which was cited hereinabove and E.U. 4,966,723 to Hodge et al., Issued October 30, 1990, includes activators of the benzoxazine type, such as the ring of CßH4 to which it is fused at positions 1, 2 a portion - C (O) OC (R1) = N-. Depending on the activator and the precise application, good bleaching results can be obtained from bleaching systems having a pH in use of from about 6 to about 13, preferably from about 9.0 to about 10.5. Typically, for example, activators with electron removal portions are used for pH scales close to neutral or subneutral. Alkalis and pH regulating agents can be used to ensure said pH. Acyllactam activators, as described in E.U. ,698,504, E.U. 5,695,679 and E.U. 5,686,014, each of which was cited hereinbefore, are very useful here, especially the acylcaprolactams (see for example WO 94-28102 A) and acyl valerolactams (see EU 5,503,639 to Willey et al., Issued April 2). , 1996 incorporated herein by reference). b) Organic peroxides, especially diacyl peroxides These are illustrated extensively in Kirk Othmer, Encyclopedia of Chemical Technology, Vol. 17, John Wiley and sons, 1982 on pages 27-90 and especially on pages 63-72, all incorporated herein by reference. If a diacyl peroxide is used, it will preferably be one that exerts a minimal adverse impact on staining / film formation. c) Blangueo catalysts containing Metal The compositions of the present invention and methods thereof can utilize metal-containing bleach catalysts that are effective for use in bleaching compositions. Preferred are bleach catalysts containing manganese and cobalt. One type of metal-containing bleach catalyst is a catalyst system comprising a transition metal cation of defined bleach catalytic activity, such as copper, iron, titanium, ruthenium, tuxethane, molybdenum, or manganese cations, a cation of auxiliary metal having little or no catalytic bleaching activity, such as zinc or aluminum cations and a scavenger having stability constants defined for catalytic and auxiliary metal cations, particularly ethylenediaminetetraacetic acid, ethylenediaminetetra (methylene phosphonic acid) and soluble salts in water of them. Said catalysts are described in E.U. 4,430,243 to Bragg, issued February 2, 1982.

Manganese Metal Complexes If desired, the compositions herein can be catalyzed by means of a manganese compound. Such compounds and levels of use are well known in the art and include, for example, the manganese-based catalysts described in the E.U. Nos. 5,576,282; 5,246,621; 5,244,594; 5,194,416; and 5.1 14,606; and European Patent Application Publication Nos. 549,271 A1, 549,272 A1, 544,440 A2, and 544,490 A1; Preferred examples of these catalysts include Mn? v2 (uO) 3 (1, 4,7-trimeti, 4,7-triazacyclononane) 2 (PF6) 2, Mnlll2 (uO) 1 (u-Oac) 2 (1, 4, 7-trimethyl-1, 4,7-triazacyclononane) 2 (CIO) 2, Mnlv (uO) 6 (1, 4,7-triazacyclononane) 4 (CIO) 4, Mn '"Mn'v4 (uO)? (u-Oac) 2- (1, 4,7-trimethyl-1, 4,7-triazacyclononane) 2 (CIO 4) 3, Mn v (1, 4,7-trimethyl-1,4,7-triazacyclononane- (OCH3) 3 (PF6), and mixtures thereof Other metal-based bleach catalysts include those described in U.S. Patent Nos. 4,430,243 and 5,164,611.The use of manganese with several complex ligands to increase or improving bleaching is also described in the following U.S. Patents: 4,728,455, 5,284,944, 5,246,612, 5,256,779, 5,280.1 17, 5,274,147, 5,153,161, and 5,227,084.

Cobalt Metal Complexes Cobalt bleach catalysts useful herein are known, and are described, for example, in US Pat. : 5,597,936; and 5,595,967; and 5,703,030; and M.L. Tobe, "Base Hydrolysis of Transition-Metal Complexes", ("Base hydrolysis of metal-transition complexes") Adv. Inorg. Bioinorg, Mexh., (1938), 2, pages 1-94. The most preferred cobalt catalyst, useful herein, is salt (s) of cobalt pentaamine acetate having the formula [Co (NH3) 5OAc] Ty, wherein "OAc" represents a portion of acetate and "Ty" is an anion, and especially cobalt pentaaminacetate chloride, [Co (NH3) 5OAc] Cl2; as well as [Co (NH3) 5? Ac] (Oac) 2; [Co (NH3) 5? Ac] (PF6) 2; [Co (NH3) 5? Ac] (SO4); [Co (NH3) 5? Ac] (BF4) 2; and [Co (NH3) 5? Ac] (NO3) 2; (in the present it is called "PAC"). These cobalt catalysts are readily prepared by known methods, such as those shown for example in the US patents. 5,597,936; 5,595,967; and 5,703,030; and in the Tobe article and the references cited therein; and in the US patent. 4,810,410; J. Chem. Ed. (1989), 66 (12), 1043-45; The Synthesis and Characterization of Inorganic Compounds, (The Synthesis and Characterization of Inorganic Compounds) W.L. Jolly (Prentice-Hall); 1970), pp. 461-3; Inorg. Chem., 18 1497-1502 (1979); Inorg. Chem., 21, 2881-2885 (1982); inorg. Chem., 18, 2023-2025 (1979); Inorg. Synthesis, 173-76 (1960); and Journal of Physical Chemistry. 56. 22-25 (1952).

Transition Metal Complexes of Macropolycyclic Rigid Ligands The compositions herein may also suitably include as a bleach catalyst, a transition metal complex of a macropolycyclic rigid ligand. The phrase "macropolycyclic rigid ligand" is sometimes abbreviated as "MRL" same as described below. The amount used is a catalytically effective amount, suitable around 1 ppb or more, for example up to about 99.9%, very typically around 0.001 ppm or more, preferably from about 0.05 ppm to about 500 ppm (where "ppb") means parts per billion in weight and "ppm" means parts per million by weight). Suitable transition metals, for example, Mn are illustrated hereinafter. "Macropolycyclic" means an MRL that is both macrocyclic and polycyclic. "Polycyclic" means at least bicyclic. The term "rigid" as used herein, here includes "having a super structure" and "joined by cross bridges". "Rigid" has been defined as the artificial inverse of flexibility: See D: H: Busch., Chemical Reviews., (1993), 93, 847-860, incorporated herein by reference. Most particularly, the term "rigid" as used herein means that the MRL must be definitely stiffer than a macrocycle ("original macrocycle") that is otherwise identical (having the same size and type of ring and number of atoms in the main ring) but lacking a superstructure (especially linkages or, preferably, cross-linked portions) found in the MRLs. To determine the comparative stiffness of macrocycles with and without superstructures, the practitioner will use the free form (not the metal link shape) of macrocycles. Rigidity is well known because it is useful for comparing macrocycles; suitable tools for determining, measuring or comparing rigidity including computer methods (see, for example, Zimmer, Chemical Reviews, (1995), 95 (38), 2629-2648 or Hancock et al., Inorganics Chimica-Acta, (1989 ), 164, 73-84 The preferred MRLs herein are a special type of ultra-rigid ligands that are linked by cross-bridges.A "cross-bridging" is illustrated in a non-limiting manner hereinafter in 1.1. In 1.1, the cross-bridge bond is a CH2CH2 portion, which links cross-bridges to N1 and N8 in the illustrative structure, by comparison, a bridge "on the same side", for example, if one were to be introduced through of N1 and N12 in 1.1 1, would not be sufficient to constitute a "crossed bridge bond" and therefore would not be preferred.The suitable metals in rigid ligand complexes include Mn (ll), Mn (III), Mn (IV ), Mn (V), Fe (ll), Fe (lll), Fe (IV), Co (l), Co (ll), Co (lll), Ni (l), Ni ( ll), Ni (lll), Cu (l), Cu (ll), Cu (lll), Cr (ll), Cr (lll), Cr (IV), Cr (V), Cr (VI), V ( lll), V (IV), V (V), Mo (IV), Mo (V), Mo (VI), W (IV), W (V), W (VI), Pd (ll), Ru ( ll), Ru (lll) and Ru (IV). Preferred transition metals in the instant transition metal bleach catalyst include manganese, iron and chromium.

Very generally, the MRLs (and the corresponding metal catalysts) herein, suitably comprise: a) at least one macrocyclic master ring comprising four or more heteroatoms; and b) a covalently connected non-metallic superstructure capable of increasing the stiffness of the macrocycle, preferably selected from i) a cross-bridge superstructure, such as a link portion: ii) a cross-bridge superstructure, such as the portion Link by crossed bridges; and iii) combinations thereof. The term "superstructure" is used herein as defined in the literature presented by Busch et al., See for example, articles by Busch in "Chemical Reviews". In the present, the preferred superstructures not only improve the stiffness of the original macrocycle, but also favor the bending of the macrocycle in such a way that it coordinates to a metal in a groove. Suitable superstructures can be extremely simple, for example a joining portion such as any of those illustrated in Figures 1 and 2 can be used.

Fig. 1 Where n is an integer, for example 2 to 8, preferably less than 6, typically from 2 to 4, or Fig 2 Where m and n are integers of about 1 to 8, most preferably 1 to 3; Z is N or CH; and T is a compatible substituent, for example H, alkyl, trialkylammonium, halogen, nitrogen, sulfonate, or the like. The aromatic ring at 1.10 can be replaced by a saturated ring, wherein the Z atom that connects within the ring can contain N, O, S or C. Suitable MRLs are further illustrated non-limitingly by the following compound: Fig. 3 This is an MRL according to the invention, which is highly preferred, crosslinked, derived from cyclamyl substituted with methyl (all tertiary nitrogen atoms). Formally, this ligand is called 5,12-dimethyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane using the extended von Baeyer system. See "A Guide to IUPAC Nomenclature of Organic Compounds: Recommendations 1993", (A Guide to the IUPAC Nomenclature of Organic Compounds: 1993 Recommendations,) by R. Panic, W.H. Powell and J-C nRicher (Eds.), Blackwell Scientific Publications, Boston, 1993; see especially section R-2.4.2.1. Metal-transition bleach catalysts of macrocyclic rigid ligands which are suitable for use in the compositions of the invention may, in general, include known compounds wherein they fit the definition herein, as well as, most preferably, any compound of a large number of novel compounds that are expressly designed for the laundry or cleaning uses present, and are illustrated in a non-limiting manner by any of the following: Dichloro-5,12-dimethyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (II) hexafluorophosphate diaquo-5,12-dimethyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (II) hexafluorophosphate aquo-hydroxy-5,12 -dimethyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (III) tetrafluoroborate diaquo-5,12-dimethyl-1, 5,8,12-tetraazabic¡clo [6.6.2] hexadecane Manganese (ll) Dichloro-5,12-dimethyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexade hexafluorophosphate gray manganese (lll) Dichloro-5,12-di-n-butyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (II) Dichloro-5,12-dibenzyl-1, 5, 8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (II) Dichloro-5-n-butyl-12-methyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexadecane Manganese (II) Dichloro 5-n-octyl-12-methyl-1, 5,8,12-tetraazabicyclo [6.6.2] hexanecane Manganese (II) Dichloro-5-n-butyl-12-methyl-1, 5,8,12- tetraazabicyclo [6.6.2] Manganese hexadecane (11) As a practical matter, and not by way of limitation, the methods of compositions and cleaning herein can be adjusted to provide in the order of at least one part per one hundred million the active bleach catalyst species in the aqueous washing medium, and preferably will provide from about 0.01 ppm to about.25 ppm, most preferably from about 0.05 ppm to about 10 ppm, and most preferably still from about 0.01 ppm to about 5 ppm, of the catalyst species of bla Locking in the washing solution. To obtain such levels in the wash solution in an automatic washing process, typical compositions herein will include from about 0.0005% to about 0.2%, most preferably from about 0.004% to about 0.08% bleach catalyst, especially manganese or cobalt catalysts, by weight of the bleaching compositions. d) Other Bleach Catalysts The compositions herein may comprise one or more different bleach catalysts. Preferred bleach catalysts are the zwitterionic bleach catalysts described in the US patent. 5,576,282 (especially propanesulfonate 3- (3,4-dihydroisoquinolinium). Other bleach catalysts include cationic bleach catalysts are described in US 5,360,569 patent, 5,442,066, 5,478,357, 5,370,826, 5,482,515, 5,550,256 and WO 95/13351, WO 95 / 13352, WO 95/13353 and. also suitable as bleaching agents preformed those killed, such as peroxy caproic phthalimido ( "PAP"). See for example US 5,487,818 patents, 5,310,934, 5,246,620, 5,279,757 and 5,132,431.

Optional Detersive Enzymes The detergent and cleaning compositions herein may optionally also contain one or more types of detergent enzymes. Said enzymes may include other proteases, amylases, cellulases and lipases. Such materials are known in the art and are commercially available under said trademarks. They may be incorporated into the non-aqueous liquid detergent compositions herein, in the form of "disk" or "pellet" suspensions. Other suitable types of enzymes comprise those in the form of enzyme suspensions in nonionic surfactants, for example, the enzymes marketed by Novo Nordisk under the tradename "SL" or the microencapsulated enzymes marketed by Novo Nordisk under the tradename "LDP " Suitable enzymes and levels of use are described in the US patents. 5,576,282, 5,705,464 and 5,710.1 15. Enzymes that are added to the compositions herein in the form of conventional enzyme pellets are especially preferred for use therein. Said pellets will generally vary in size from about 100 to 1000 microns, most preferably from about 200 to 800 microns and may be suspended throughout the non-aqueous liquid phase of the composition. It has been found that the pellets in the compositions of the present invention, in comparison with other forms of enzymes, exhibit primarily the desirable enzyme stability in terms of retention of enzymatic activity as time elapses. Therefore, compositions using enzyme pellets do not need to contain the conventional enzyme stabilizer such as would be used very frequently when the enzymes are incorporated in aqueous liquid detergents. However, the enzymes that are added to the compositions herein can be granulated, preferably T-granulated. "Detersive Enzyme", as used herein, means any enzyme that has a cleaning, stain removal or otherwise beneficial effect in a detergent composition for laundry, hard surfaces, cleaning or personal care. Preferred detersive enzymes are hydrolases such as proteases, amylases and lipases. Preferred enzymes for laundry purposes include, but are not limited to, proteases, cellulases, lipases and peroxidases. Amylases and / or proteases are highly preferred for automatic dishwashing, including both current commercially available types and improved types, which, although they are more and more compatible with bleaching although they have successive improvements, still retain a residual degree of susceptibility to deactivation of bleaching. Examples of suitable enzymes include, but are not limited to hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, keratanases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tanases, pentosanas, malanases , known ß-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase and amylases, or mixtures thereof. Examples of such suitable enzymes are described in the E.U. Nos. 5,705,464, 5,710.1 15, 5,576,282, 5,728,671 and 5,707,950. Cellulases useful in the present invention include both bacterial and fungal cellulases. Preferably, they will have an optimum pH between 5 and 12 and a specific activity above 50 CEVU / mg (Cellulase Viscosity Unit). Suitable cellulases are described in the U.S. Patent. 4,435,307, J61078384 and WO95 / 26398 which describes a fungal cellulase produced respectively from Humicola insolens, Trichoderma, Thielavia and Sporotrichum. EP 739 982 describes cellulases isolated from the novel species Bacillus. Suitable cellulases are also described in GB-A-2,075,028; GB-A-2,095,275, DE-OS-2,247,832 and WO95 / 26398. Examples of said cellulases are cellulases produced by a strain of Humicola insolens (Humicola grisea var. Thermoidea), particularly the strain Humicola DSM 1800. Other suitable cellulases are cellulases produced by a strain of Humicola insolens, which has a molecular weight of about 50 KDa, an isoelectric point of 5.5. and it contains 415 amino acids; and an endoglucanase -43 kD derived from Humicola insolens, DSM 1800, exhibiting cellulase activity; a preferred endoglucanase component having the amino acid sequence described in WO91 / 17243. Also suitable are the EGIII cellulases of Trichoderma longibrachiatum described in WO94 / 21801 to Genencor. Especially suitable cellulases are cellulases that have color care benefits. Examples of said cellulases are cellulases which are described in European Patent Application No. 91202879.2 filed on November 6, 1991 (Novo). Carezyme and Celluzyme (Novo Nordisk A / S) are especially useful. See also WO91 / 17244 and WO91 / 21801. Other suitable cellulases with fabric care and / or cleaning properties are described in WO96 / 34092, WO96 / 179994 and WO95 / 24471. Said cellulases, when present, are normally incorporated in the cleaning composition in levels from 0.0001% to 2% pure enzyme by weight of the cleaning composition. Peroxidase enzymes are used in combination with oxygen sources, for example, percarbonate, perborate, persulfate, hydrogen peroxide, etc., and with a phenolic substrate as a bleach-enhancing molecule. They are used for "bleaching in solution", that is, to avoid the transfer of dyes or pigments removed from substrates during washing operations, to other substrates in the washing solution. Peroxidase enzymes are known in the art and include, for example, horseradish peroxidase, ligninase, and haloperoxidase such as chloro- and bromo-peroxidase. Peroxidase-containing detergent compositions and the peroxidases themselves are described, for example, in US Pat. Nos. 5,705,464, 5,710.1 15, 5,576,282, 5,728,671 and 5,707,950, PCT International Application WO89 / 099813, WO 89/09813 and in European Patent Application No. 91202882.6, filed November 6, 1991 and EP No. 96870013.8, filed on February 20, 1996. Also the laccase enzyme is adequate. The increments are generally constituted at a level of 0.1% to 5% by weight of the total composition. Preferred enhancers are fentiazine and phenoxasine, 10-phenothiazinopropionic acid (PPT), 10-ethylphenothiazine-4-carboxylic acid (EPC), 10-phenoxazinopropionic acid (POP) and 10-methylphenoxazine (described in WO 94/12621) and substituted syringes (substituted C3-C5 alkylsalicylates) and phenols. Percarbonate or sodium perborate are preferred sources of hydrogen peroxide. Said peroxidases are normally incorporated in the cleaning composition at levels from 0.0001% to 2% active enzyme by weight of the cleaning composition. Enzymatic systems can be used as bleaching agents. Hydrogen peroxide may also be present by adding an enzymatic system (i.e., an enzyme and a substrate) that has the ability to generate hydrogen peroxide at the beginning or during the washing and / or rinsing process. Said enzymatic systems are described in the European patent application 91202655.6 filed on October 9, 1991. Other preferred enzymes that can be included in the cleaning compositions of the present invention include lipases. Suitable lipase enzymes for detergent use include those produced by microorganisms of the Pseudomonas group, such as Pseudomonas stutzeri ATCC 19,154, such as those described in British Patent 1, 372,034. Suitable lipases include those that show a positive immunological cross-reaction with the lipase antibody, produced by the microorganism Pseudomonas fluorescent IAM 1057. This lipase is available from Amano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name Lipasa P "Amano", which in the future is called "Amano-P". Other suitable commercial lipases include Amano-CES, lipases ex Chromobacter viscosum, for example Chromobacter viscosum var. lipolyticum NRRLB 3673, from Toyo Jozo Co., Tagata, Japan; Chromobacter viscosum lipases from U.S. Biochemical Corp, E.U. and Disoynth Co., The Netherlands and lipases ex Pseudomonas gladioli. Especially suitable lipases are lipases such as M1 Lipase® and Lipomax® (Gist-Brocades) and Lipolase® and Lipolase Ultra® (Novo), which have been found to be very effective when used in combination with the compositions of the present invention. Also suitable are the lipolytic enzymes described in EP 258 068, WO 92/05249 and WO 95/22615 by Novo Nordisk, and in WO 94/03578, WO 95/35381 and WO 96/00292 by Unilever. Also suitable are cutinases [EC 3.1.1 .50] that can be considered as a special type of lipase, mainly lipases that do not require interfacial activation. The addition of cutinases to cleaning compositions has been described, for example, in: WO-A-88/09367 (Genencor); WO 90/09446 (Plant Genetic System), and WO 94/14963 and WO 94/14964 (Unilever).

Lipases and / or cutinases, when present, are normally incorporated in the detergent composition at levels of 0.0001% to 2% active enzyme by weight of the detergent composition. In addition to the lipases mentioned above, the phospholipases can be incorporated into the cleaning compositions of the present invention. Non-limiting examples of suitable phospholipases include: EC 3.1 .1.32 Phospholipase A1; EC 3.1.1.4 Phospholipase A2; EC 3.1.1.5 Lisofolipase; EC 3.1.4.3 Phospholipase C; EC 3.1.4.4. Phospholipase D. Commercially available phospholipases include LECITASE ® from Novo Nordisk A S from Denmark and Phospholipase A2 from Sigma. When phospholipases are included in the compositions of the present invention, it is preferable that amylases are also included. Without wishing to be limiting by theory, the combined action of phospholipase and amylase is considered to provide substantive spot removal, especially in greasy / oily, starchy and highly nuanced spots and dirt. Preferably, the phospholipase and the amylase, when present, are incorporated in the compositions of the present invention at a weight ratio of pure enzyme between 4500: 1 and 1: 5, most preferably between 50: 1 and 1: 1. Suitable proteases are the subtilisins that are obtained from particular strains of B.subtilis and B.licheniformis (subtilisin BPN and BPN '). A suitable protease is obtained from a Bacillus strain, which has a maximum activity across the pH range of 8 to 12, which is developed and sold as ESPERASE® by Novo Industries A / S of Denmark, hereinafter "Novo" . The preparation of this enzyme and analogous enzymes is described in GB 1, 243,784, by Novo. Proteolytic enzymes also encompass modified bacterial serine proteases, such as those described in European Patent Application No. 87303761.8, filed April 28, 1987 (particularly pages 17, 24 and 98) same as herein referred to as " Protease B ", and in the European patent application EP 199 404, Venegas, published on October 29, 1986, which refers to a modified bacterial serine proteolytic enzyme that is referred to herein as" Protease A ". "Protease C" is suitable as it is referred to herein, which is a variant of a Bacillus alkaline serine protease in which lysine replaces arginine at position 27, tyrosine replaces valine at position 104, serine replaces asparagine at position 123 and alanine replaces threonine at position 274. Protease C is described in EP 90915958.4, corresponding to WO 91/06637, published May 16, 1991. Also included herein are genetically modified variants, particularly of protease C. A preferred protease which is referred to as "Protease D" is a variant of carbonyl hydrolase as described in the EU patent. No. 5,677,272, and WO95 / 10591. Also suitable is a carbonyl hydrolase variant of the protease described in WO95 / 10591, which has an amino acid sequence derived by the replacement of a plurality of amino acid residues replaced in the precursor enzyme at position +210, in combination with one or more residuals: +33, +62, +67, +76, +100, +101, +103, +104, +107, +128, +129, +130, +132, +135, +156, + 158, +164, +166, +167, +170, +209, +215, +217, +218 and +222 where the numbered position corresponds to the subtilisin that occurs naturally from Bacillus amyloliquefaciens or to residues s equivalent amino acids in other carbonyl hydrolases or subtilisins, such as subtilisin Bacillus lentus (co-pending US patent application Serial No. 60 / 048,550, filed on June 4, 1997 and PCT international application Serial No. PCT / IB98 / 00853.) suitable for the present invention are the proteases described in patent applications EP 251 446 and WO 91/06637 and the BLAP® protease described in WO91 / 02792 and its variants described in WO 95/23221. See also a high pH protease from Bacillus sp. NCIMB 40338 described in WO 93/18140 A to Novo. Enzymatic detergents comprising protease, one or more other different enzymes and a reversible protease inhibitor are described in WO 92/03529 A to Novo. When desired, a protease having decreased adsorption and increased hydrolysis is available as described in WO 95/07791 to Procter & amp;; Gamble. A recombinant trypsin-like protease for detergents suitable herein is described in WO 94/25583 to Novo. Other suitable proteases are described in EP 516 200 by Unilever. Particularly useful proteases are described in the publications: WO 95/30010; WO 95/3001 1; and WO 95/29979. Suitable proteases are commercially available as ESPERASE® ALCALASE®, DURAZYM®, SAVINASE®, EVERLASE® and KANNASE®, all of Novo Nordisk A / S from Denmark, and as MAXATASE®, MAXACAL®, PROPERASE® and MAXAPEM® all of them of Genencor International (formerly Gist-Brocades of the Netherlands). Said proteolytic enzymes, when present, are incorporated in the cleaning compositions of the present invention at a level of from 0.0001% to 2%, preferably from 0.001% to 0.2%, most preferably from 0.005% to 0.1% pure enzyme by weight of the total composition. Amylases (a and / or ß) can be included for the removal of carbohydrate-based spots. WO94 / 02597, describes cleaning compositions that incorporate mutant amylases. See also WO95 / 10603, Other amylases known to be used in cleaning compositions include α- and β-amylases. A-amylases are known in the art and include those described in US Patent Nos. 5,003,257, EP 252,666; WO / 91/00353; FR 2,676,456; EP 285,123; EP 5235,610; EP 368,341 and British Patent Specification No. 1, 296,839 (Novo). Other suitable amylases are amylases with increased stability which are described in WO94 / 18314, and WO96 / 05295, Genencor, and the amylase variants having further modification in their immediate similar available from Novo Nordisk A / S which is described in WO95 / 10603 . Also suitable are the amylases described in EP 277 216. Examples of commercial α-amylase products are Purafect Ox Am® from Genencor and Termamyl®, Ban®, Fungamyl® and Duramyl® all available from Novo Nordisk A / S Denmark. WO95 / 26397 describes other suitable amylases: α-amylases which are characterized by having a specific activity at least 25% higher than the specific activity of Termamyl® at a temperature range of 25 ° C to 55 ° C at a pH value in the scale from 8 to 10, measured by the Phadebas® α-amylase activity test. The variants of the above enzymes are suitable, which are described in WO96 / 23873 (Novo Nordisk). Other amylolytic enzymes with improved properties with respect to their activity level and the combination of thermostability and a higher activity level are described in WO95 / 35382. Said amylolytic enzymes, when present, are incorporated in the cleaning compositions of the present invention at a level of from 0.0001% to 2%, preferably from 0.00018% to 0.06%, most preferably from 0.00024% to 0.048% pure enzyme by weight of the composition. The aforementioned enzymes may have any suitable origin, such as vegetable, animal, bacterial, fungal and yeast. The origin can also be mesophilic or extremophile (psychrophilic, psychrotrophic, thermophilic, barophilic, alkalophilic, acidic, halophilic, etc.). The purified or non-purified forms of these enzymes can be used. In these days it is a common practice to modify wild-type enzymes by means of genetic manipulation or protein techniques to optimize their efficiency of performance in the cleaning compositions of the invention. For example, the variants can be designed in such a way that the compatibility of the enzyme with the ingredients of said commonly found compositions is increased. Alternatively, the variant can be designed such that the optimum pH, bleach or chelator stability, catalytic activity and the like of the enzyme variant are designed to conform to the particular cleaning application. In particular, attention should be focused on amino acids sensitive to oxidation in the case of bleach stability and on surface charges for compatibility with the surfactant. The isoelectric point of these enzymes can be modified by replacing some charged amino acids, for example, an increase in the isoelectric point could help improve compatibility with anionic surfactants. The stability of the enzymes can be further increased by the creation of additional salt bridges for example, and by reinforcing the calcium binding sites to increase the stability in the chelator. These optional detersive enzymes, when present, are normally incorporated in the cleaning composition at levels of 0.0001% to 2% pure enzyme by weight of the detergent composition. Enzymes may be added as separate individual ingredients (pellets, granules, stabilized liquids, etc. containing an enzyme) or as mixtures of two or more enzymes (eg cogranulates).

Other suitable detergent ingredients that may be added are the enzyme oxidation scavengers. Examples of said enzyme oxidation scavengers are ethoxylated tetraethylenepolyamines. A range of enzyme materials and means for their incorporation into synthetic detergent compositions is also disclosed in WO 9307263 and WO 9307260 to Genencor International, WO 8908694 and E.U. 3,553,139, January 5, 1971 to McCarty and others. Enzymes are also described in E.U. 4,101, 457, and in E.U. 4,507,219. Useful enzyme materials for liquid detergent formulations and their incorporation into such formulations are described in E.U. 4,261, 868.

Enzyme Stabilizers The enzymes that will be used in detergents can be stabilized by various techniques. Enzyme stabilization techniques are described and exemplified in E.U. 3,600,319, and EP 199,405 and EP 200,586. Enzyme stabilization systems are also described, for example, in E.U. 3,519,570. A Bacillus sp. AC13 useful and giving proteases, xylanases and cellulases is described in WO 9401532. The enzymes employed herein can be stabilized by the presence of water soluble sources of calcium and / or magnesium ions in the finished compositions, which provide said ions to the enzymes. Suitable enzyme stabilizers and their levels of use are described in US Patent Nos. 5,705,464, 5,710.1, 15 and 5,576,282.

Detergency builders The detergent and cleaning compositions described herein preferably comprise one or more detergency builders or builders. When present, the compositions will typically comprise at least about 1% builder, preferably about 5%, most preferably about 10% to about 80%, preferably about 50%, most preferably about 30% by weight, of the detergency builder. However, it does not mean that lower or higher levels of the detergency builder are excluded. Preferred builders for use in detergent and cleaning compositions, particularly in dishwashing compositions, described herein include, but are not limited to, water-soluble builder compounds (e.g. carboxylates) as was described in the EU patents Nos. 5,695,679; 5,705,464 and 5, 710.1 15. Other suitable polycarboxylates are described in the E.U. Nos. 4,144,226; 3,308,067 and 3,723,322. Preferred polycarboxylates are hydroxycarboxylates containing up to three carboxy groups per molecule, very particularly titrates. Inorganic or P-containing builders include, but are not limited to, the alkali metal, ammonium and alkanolammonium polyphosphate salts (which are polymerized tripolyphosphates, pyrophosphates, and polymeric meta-phosphates), phosphonates (see for example, US Patent Nos. 3,159,581, 3,213,030, 3,422,021, 3,400,148 and 3,422,137), phytic acid, silicates, carbonates (including bicarbonates and sesquicarbonates), sulfates, and aluminosilicates. However, non-phosphate builders are required in some cases. It is very important that the compositions herein work surprisingly well even in the presence of the "weak" builders (compared to the phosphates) such as citrate, or in the so-called "under preparation" situation which may occur with the zeolite builders or layered silicates. Suitable silicates include the water-soluble sodium silicates with a SiO2: Na2O ratio of about 1.0 to 2.8, with ratios of about 1.6 to 2.4 as most preferred, and about a ratio of 2.0, which is the most preferred The silicates can be in any form, either anhydrous salt or a hydrated salt. Sodium silicate with a SiO2: Na2O ratio of 2.0, which is the most preferred. The silicates, when present, preferably present in the detergent and cleaning compositions described herein, at a level of from about 5% to about 50% by weight of the composition, most preferably from about 10% to about 40% in weigh. The partially soluble or insoluble builder compounds, which are suitable for use in detergent and cleaning compositions, particularly in granular detergent compositions, include, but are not limited to, crystalline layered silicates, preferably crystalline layered sodium silicates. , (partially soluble in water) as described in the EU patent No. 4,664,839, and sodium aluminosilicates (insoluble in water). When present in the detergent and cleaning compositions, these builders are typically present at a level of from about 1% to 80% by weight, preferably about 10% to 70% by weight, most preferably about 20% by weight. % to 60% by weight of the composition. The crystalline layered sodium silicates have the general formula NaMSix? 2? + 1.yH2O wherein M is sodium or hydrogen, x is a number from about 1.9 to about 4, preferably from about 2 to about 4, most preferably 2 , e and is a number from about 0 to about 20, preferably, 0 can be used in the compositions described herein. The crystallized statified sodium silicates of this type are described in EP-A-0164514 and the methods for their preparation are described in DE-A3417649 and DE-A-3742043. The most preferred material is delta-Na2SiO5, available from Hoechst AG as NaSKS-6 (which is commonly abbreviated here as "SKS-6"). Unlike zeolite builders, the NaSKS-6 silicate builder does not contain aluminum. NaSKS-6 has the morphological form delta-Na2SiO5 of layered silicate. SKS-6 is a highly preferred layered silicate for use in the compositions described herein, but other determined layered silicates, such as those having the general formula NaMSixO2x + 1.yH2O wherein M is sodium or hydrogen, x is a number from 1.9 to 4, preferably 2, and y is a number from 0 to 20, preferably 0 can be used in the compositions described herein. Other various layered silicates available from Hoechst include NaSKS-5, NaSKS-7 and NaSKS-1 1, such as the alpha, beta and gamma forms. As indicated above, delta-Na2S¡O5 (NaSKS-6 form) is most preferred for use herein. Other silicates such as for example magnesium silicate, which may be useful as a tightening agent in granulated formulations, as a stabilizing agent for oxygen bleaches, and as a component of foam control systems may also be useful. The crystalline layered sodium silicate material is preferably present in granular detergent compositions as a particulate ingredient in an intimate mixture with a solid ionisable water-soluble material. The water-soluble solid ionizable material is preferably selected from organic acids, organic and inorganic acid salts and mixtures thereof. Aluminosilicate builders are of great importance in heavy duty granular detergent compositions that are very commonly marketed on the market today, and can also be a significant detergency enhancing ingredient in liquid detergent formulations. The aluminosilicate builders have the empirical formula: [M2 (Ai02) and]. XH2O where z and e are integers of at least 6, the molar ratio of z and e is on the scale of 1.0 to about 0.5, and x is an integer of about 15 to about 264. Preferably the aluminosilicate builder is an aluminosilicate zeolite having the unit cell formula: Na2 [AIO2) z (SiO2) and] .xH2O where z and e are at least 6; the mole ratio of zey is from 1.0 to 0.5 and x is at least 5, preferably 7.5 to 276, most preferably from 10 to 264. The aluminosilicate builders are preferably in hydrated form and are preferably crystalline, containing about 10. % to about 28%, most preferably from about 18% to about 22% water in bound form. These aluminosilicate ion exchange materials can be crystalline to amorphous in structure and can be aluminosilicate which occurs naturally or synthetically derived. A method for producing aluminosilicate ion exchange materials is described in the U.S. patent. 3,985,669. The preferred synthetic crystalline aluminosilicate ion exchange materials useful herein are available under the names Zeolite A, Zeolite B, Zeolite P, Zeolite X, Zeolite MAP and Zeolite HS and mixtures thereof. In an especially preferred embodiment, the crystalline aluminosilicate ion exchange material has the formula: Na12 [(AIO2) 22 (SiO2) 12] -xH20 wherein x is about 20 about 30, especially about 27. This material is known as Zeolite A. Dehydrated zeolites (x = 0-10) can also be used in the present. Preferably, the aluminosilicate has a particle size of about 0.1-10 microns in diameter. The Zeolite X, has the formula: Na86 [AIO2) 86 (S¡O2) 106] -276H2O The citrate builders, for example citric acid and soluble salts thereof (particularly sodium salt), are builders of detergency. polycarboxylate of particular importance for liquid heavy duty detergent formulations due to its availability from renewable resources and due to its biodegradability. The citrates can also be used in granular compositions, especially in combination with zeolite builders and / or layered silicate. Oxydisuccinates are also especially useful in such compositions and combinations. Also suitable in the detergent compositions described herein are the 3,3-dicarboxy-4-oxa-1,6-hexanedioates and the related compounds described in E.U. 4,566,984. Useful succinic acid builders include the C5-C20 alkyl succinic and alkenisuccinic acids and salts thereof. A particularly preferred compound of this type is dodecenylsuccinic acid. Specific examples of succinate builders include: lauryl succinate, myristiisuccinate, palmitylsuccinate, 2-dodecenylsuccinate (most preferred), 2-pentadecenylsuccinate and the like. Lauryl succinates are the preferred builders of this group, and are described in European Patent Application 86200690.5 / 0,200,263, published on November 5, 1986. Fatty acids, for example, C 2-2 monocarboxylic acids, can also be incorporated. C? 8, in the compositions, alone or in combination with the builders mentioned above, especially citrate and / or succinate builders, to provide an additional builder activity. Such use of fatty acids will generally result in a decrease in foam capacity, which must be taken into account by the formulator.

Dispersants One or more suitable polyalkyleneimine dispersants may be incorporated in the cleaning compositions of the present invention. Examples of such suitable dispersants can be found in European Patent Application Nos .: 1 1 1, 965, 1 1 1, 984, and 1 12,592; and in the US patents. Nos .: 4,597,898; 4,548,744, and 5,565,145. However, any suitable dispersing or anti-redeposition agent of clays / soils can be used in the laundry compositions of the present invention. In addition, polymeric dispersing agents including polymeric polycarboxylates and polyethyleneglycols, are suitable for use in the present invention. The unsaturated monomeric acids which can be polymerized to form suitable polymeric polycarboxylates include acrylic acid, maleic acid (or maleic anhydride), fumaric acid, taconic acid, aconitic acid, mesaconic acid, citraconic acid and methylenemalonic acid. Particularly suitable polymeric polycarboxylates can be acrylic acid derivatives. Said acid-based acrylic polymers which are useful herein are the water-soluble salts of polymerized acrylic acid. The average molecular weight of such polymers in the acid form preferably ranges from about 2,000 to 10,000, most preferably from about 4,000 to 7,000, and most preferably even from about 4,000 to 5,000. The water-soluble salts of said acidic acrylic polymers may include, for example, the alkali metal, ammonium and substituted ammonium salts. Soluble polymers of this type are known materials. For example, the use of polyacrylates of this type in detergent compositions has been described, for example in E.U. 3,308,067. The acrylic / maleic acid base copolymers can also be used as a preferred component of the dispersing / redeposition agent. Such materials include the water-soluble salts of copolymers of acrylic acid and maleic acid. The average molecular weight of said copolymers in the acid form preferably ranges from about 2,000 to 10,000 more preferably from about 5,000 to 75,000, most preferably even from about 7,000 to 65,000. The ratio of acrylate to maleate segments in said copolymers will generally vary from about 30: 1 to about 1: 1, most preferably from about 10: 1 to 2: 1. The water-soluble salts of said acrylic acid / maleic acid copolymers may include, for example, the alkali metal, ammonium and substituted ammonium salts. Soluble acrylate / maleate copolymers of this type are known materials which are described in European Patent Application No. 66915, published on December 15, 1982, as well as in EP 193,360, published September 3, 1986, which also describes such polymers including hydroxypropyl acrylate. Dispersing agents that are also useful include the maleic / acrylic / vinyl alcohol terpolymers. Such materials are also described in EP193,360, including, for example, the terpolymer 45/45/10 of acrylic / maleic / vinyl alcohol. Another polymeric material that can be included is polyethylene glycol (PEG). PEG can exhibit the performance of dispersing agent as well as act as a removal and antiredeposition agent of soils and clays. The typical molecular weight varies for these purposes from about 500 to about 100,000, preferably from about 1,000 to about 50,000, and most preferably from about 1,500 to about 10,000.

Dispersants of polyaspartate and polyglutamate, especially together with zeolite builders, can also be used. Dispersing agents such as polyaspartate preferably have a molecular weight (average) of about 10,000.

Agents that release dirt The compositions according to the present invention may optionally comprise one or more agents that release dirt. If they are used, these agents that release dirt will generally comprise about 0.01%, preferably about 0.1%, most preferably from about 0.2% to about 10%, preferably about 5%, and most preferably even about 3% by weight of the composition. Non-limiting examples of polymers that release suitable soils are described in the US patents. Nos .: 5,728,671; 5,691, 298; 5,599,782; 5,415,807; 5,182,043; 4,956,447; 4,976,879; 4,968,451; 4,925,577; 4,861, 512; 4,877,896;, 4,771, 730; 4.71 1, 730; 4,721, 580; 4,000,093; 3,959,230; and 3,893,929; and European Patent Application 0 219 048. Agents releasing suitable soils are also described in US Patents. Nos. 4,201, 824; 4,240,918; 4,525,524; 4,579,681; 4,220,918; and 4,787,989; EP 279,134 A; EP 457,205 A; and DE 2,335,044.

Chelating Agents The detergent compositions herein may also contain a chelating agent useful for chelating metal ions and metal impurities that would otherwise tend to deactivate the bleaching agent (s). Useful chelating agents can include aminocarboxylates, phosphonates, aminophosphonates, polyfunctionally substituted aromatic chelating agents and mixtures thereof. Additional examples of suitable chelating agents are described in the US patents. Nos. 5,705,464, 5,710.1 15, 5,728,671 and 5,576,282. The compositions herein may also contain water-soluble salts of methyl glycine diacetic acid (MGDA) (or acid form) as a useful chelator or co-builder with, for example, insoluble builders such as zeolites, layered silicates and the like. If used, these chelating agents should generally comprise from about 0.1% to about 15% by weight of the detergent compositions herein.

Foam suppressor Another optional ingredient is a foam suppressor exemplified by silicones and silica-silicone blends. Examples of suitable foam suppressors are described in the U.S. Nos. 5,707,950 and ,728,671. These foam suppressors are normally employed at levels from 0.001% to 2% by weight of the composition, preferably from 0.01% to 1% by weight.

Softening agents Fabric softening agents can also be incorporated into laundry detergent compositions in accordance with the present invention. Inorganic softening agents are exemplified by the smectite clays described in GB-A-1 400 898 and in the US patent. No. 5,019,292. Organic fabric softening agents include water-insoluble tertiary amines such as those described in GB-A1 514 276 and EP-BO 01 1 340 and their combination with C12-C14 monoquaternary ammonium salts are described in EP-BO 026 527 and EP-BO-026 528 and the long chain diamides as described in EP-BO 242 919. Other useful organic ingredients of fabric softening systems include the high molecular weight polyether oxide materials as described in EP-AO 299 575 and 0 313 146. Particularly suitable agents are described in the EU patents Nos. 5,707,950 and 5,728,673. Smectite clay levels are usually in the range of 2% to 20%, most preferably 5% to 15% by weight, the material is added either spray-dried or dry-mixed to the rest of the formulation. Organic fabric softening agents such as water-soluble tertiary amines or long-chain amide materials are incorporated at levels of 0.5% to 5% by weight, usually from 1% to 3% by weight, while the materials of High molecular weight polyethylene oxide and water soluble cationic materials are added at levels from 0.1% to 2%, usually from 0.15% to 1.5% by weight. These materials are normally added to the portion of the composition that has been spray dried, although in some cases it may be more convenient to add them as a dry-mixed particulate material, or sprinkle them as a molten liquid over the other solid components of the composition . The biodegradable quaternary ammonium compounds as described in EP-A-040 562 and EP-A-239 910 have been presented as alternatives for the traditional long-chain alkylammonium chlorides and methylsulfates. Non-limiting examples of anions compatible with the softener for the quaternary ammonium compounds and amine precursors include chlorine or methylisulfate.

Inhibition of dye transfer The detergent compositions of the present invention can also include compounds for inhibiting the transfer of dyes from one fabric to another, of solubilized and suspended dyes encountered during fabric washing operations including dyed fabrics.

Polymeric Dye Transfer Inhibitory Agents Laundry detergent compositions according to the present invention also comprise from 0.001% to 10%, preferably 0.01% to 2%, most preferably from 0.05% to 1% by weight of transfer inhibiting polymeric agents of dyes. Said polymeric dye transfer inhibiting agents are normally incorporated into the detergent compositions to inhibit the transfer of dyes from the dyed fabrics onto the fabrics washed with the same compositions. These polymers have the ability to complex with or adsorb washed fugitive dyes from dyed fabrics before the dyes have an opportunity to bind to other articles in the wash. Especially suitable dye transfer inhibiting polymeric agents are polymers of polyamine N-oxides, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polymers of polyvinylpyrrolidone, polyvinyloxazolidones, polyvinylimidazolones and mixtures thereof. Examples of such transfer inhibiting agents are described in the U.S. Nos. 5,707,950 and 5,707,951. Additional suitable transfer inhibiting agents include, but are not limited to, entangled polymers. The entangled polymers are polymers whose base structures are interconnected to a certain degree; these links can be of a chemical or physical nature, possibly with active groups in the base structure or on the ramifications; the entangled polymers have been described in the Journal of Polymer Science, volume 22, pages 1035-1039. In one embodiment, the entangled polymers are made in such a way that they form a rigid three-dimensional structure that can trap dyes in the pores formed by the three-dimensional structure. In another embodiment, the entangled polymers trap the dyes by swelling. Said entangled polymers are described in European co-pending patent application 94870213.9 The addition of said polymers also increases the yield of the enzymes according to the invention.

PH and Variation of pH Regulation Many of the detergent and cleaning compositions described herein will be pH regulated, i.e. they are relatively resistant to pH drop in the presence of acid solids. However, other compositions herein may have an exceptionally low pH regulation capacity, or may be substantially free of pH regulation. The techniques to control or vary the pH to the recommended levels of use very generally include the use not only of pH regulators, but also of additional alkalis, acids, systems of sudden variation of pH, containers of double compartment, etc. they are well known to those in the art. Preferred ADD compositions herein, comprise a pH adjusting component selected from water soluble alkaline inorganic salts and water soluble organic and inorganic builders, as described in US Pat. Nos. 5,705,464 and 5,710.1 15.

Material Care Agents Preferred ADD compositions may contain one or more agents for the care of materials that are effective as corrosion inhibitors and / or anti-mold assistants as described in US Pat. Nos. 5,705,464, 5,710.1 15 and 5,646,101. When present, said protective materials are preferably incorporated at lower levels, for example, from about 0.01% to about 5% of the ADD composition.

Other materials The detersive or auxiliary ingredients optionally included in the instant compositions may include one or more materials to assist or increase the cleaning performance, the treatment of the substrate to be cleaned, or, are designed to improve the aesthetics of the compositions . The auxiliary ingredients may also be included in the compositions of the present invention, at their levels established in the conventional art, to be used (generally, the auxiliary materials comprise in total, from about 30% to about 99.9%, preferably from about 70% to about 95% by weight of the compositions), include other active ingredients such as phosphate builders, anti-stain ingredients, silver care, anti-mildew and / or anti-corrosion, colorants, fillers, germicides , alkalinity sources, hydrotropes, antioxidants, perfumes, solubilizing agents, vehicles, processing aids, pigments and pH control agents as described in the EU patents Nos. 5,705,464, 5,710.1 15, 5,698,504, 5,695,679, 5,686,014 and 5,646,101.

Washing Methods In addition to the methods for cleaning fabrics, tableware and other hard surfaces, as well as the body by personal cleansing, which are described herein, this invention also encompasses a laundry pre-treatment procedure for fabrics that have been soiled or stained, same comprising directly contacting said stains and / or soils with a highly concentrated form of the cleaning composition that is established above, prior to washing said fabrics using conventional aqueous washing solutions. Preferably, the cleaning composition is kept in contact with the dirt / stain for a period of about 30 seconds to 24 hours prior to the conventional washing of previously soiled / stained substrate. Most preferably, the pretreatment times will vary from about 1 to 180 minutes. The following examples are designed to exemplify compositions of the present invention, but are not necessarily designed to limit or otherwise define the scope of the invention. In all of the following examples, Protease 1 means a protease variant comprising the substitution of amino acid residues with another amino acid residue that occurs naturally at the positions corresponding to positions 101 G / 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K of subtilisin Bacillus amyloliquefaciens. Protease 1 can be substituted with any other additional protease variant of the present invention, with substantially similar results in the following examples. In the examples of the detergent compositions of the present invention, the Protease 1 enzyme levels are expressed as pure enzyme by weight of the total composition, the other enzyme levels are expressed as starting material by weight of the total composition, and less to be specified otherwise, the other ingredients are expressed by weight of the total composition. Additionally, some abbreviations known to those skilled in the art are used in the following examples, which are adapted to the description set forth herein.

Examples of cleaning compositions for hard surfaces. tableware and fabrics 1 .- Compositions for cleaning hard surfaces As used herein, a "hard surface cleaning composition" refers to liquid and granular detergent compositions for cleaning hard surfaces such as floors, walls, bathroom tiles and the like. . The hard surface cleaning compositions of the present invention comprise an effective amount of one or more protease enzymes, preferably from about 0.0001% to about 10%, most preferably from about 0.001% to about 5%, most preferably still from 0.001% to about 1% by weight of active protease enzyme of the composition. In addition to comprising one or more protease enzymes, said hard surface cleaning compositions comprise a surfactant and a water-soluble sequestering detergent builder. However, in certain specialized products such as glass aerosol cleaners, surfactants are sometimes not used since they can produce a residue that forms a scratched film / effect on the surface of the glass. (See U.S. Patent No. 5,679,630 in the examples). The surfactant component, when present, may include as little as 0.1% of the compositions herein, but typically the compositions will contain from about 0.25% to about 10%, most preferably from about 1% to about 5% of the surfactant. Typically the compositions will contain from about 0.5% to about 50% of a builder, preferably from about 1% to about 10%. Preferably, the pH should be in the range of about 8 to 12. Conventional pH adjusting agents such as sodium hydroxide, sodium carbonate or hydrochloric acid can be used if an adjustment is necessary. Solvents may be included in the compositions. Useful solvents include, but are not limited to, glycol ethers such as diethylene glycol monohexyl ether, diethylene glycol monobutyl ether, ethylene glycol monobutyl ether, ethylene glycol monohexyl ether, propylene glycol monobutyl ether, dipropylene glycol monobutyl ether, and diols such as methanol. , 2,4-trimethyl-1,3-pentanediol and 2-ethyl-1,3-hexanediol. When used, said solvents are typically present at levels of from about 0.5% to about 15%, preferably from about 3% to about 11%. Additionally, highly volatile solvents such as isopropanol or ethanol can be used in these compositions to facilitate a more rapid evaporation of the composition, from the surfaces when the surface is not rinsed after application of "full strength" of the composition to the surface. When used, volatile solvents are typically present at levels of from about 2% to about 12% in the compositions. The hard surface of the cleaning composition body of the present invention is illustrated by the following non-limiting examples.

EXAMPLES 1 -7 SL Clean Surface Hard Cleaning Compositions Example No. Component 1 2 3 4 5 6 7 Protease 1 0.05 0.05 0.20 • 0.02 0.03 0.10 0.03 Protease 2 - - - - - 0.20 0.1 Chelator ** - - - 2.90 2.90 - - Citrate - - - - - 2.90 2.90 LAS - 1 .95 - 1.95 - 1 .95 - AS 2.00 - 2.20 - 2.20 - 2.20 AES 2.00 - 2.20 - 2.20 - 2.20 Amine oxide 0.40 - 0.50 - 0.50 - 0.50 Hydrotrope - 1 .30 - 1.30 - 1.30 - Solvent *** - 6.30 6.30 6.30 6.30 6.30 6.30 Water and minor ingredients Rest at 100% 2 Protease other than Protease 1 including but not limited to proteases useful in the present invention described herein. ** Ethylenediamine diacetic acid Na4 *** Diethylene glycol monohexyl ether **** All formulas adjusted to pH 7 In Examples 6 and 7, any combination of the protease enzymes useful in the present invention are cited here, inter alia, they are replaced by Protease1 and Protease2, with substantially similar results.

EXAMPLES 8-13 Aerosol compositions for cleaning hard surfaces v removal of domestic mold Example No. Component 8 9 10 11 12 13 Protease 0.20 0.05 0.10 0.30 0.20 0.30 Protease2 - - - - 0.30 0.10 C8AS 2.00 2.00 2.00 2.00 2.00 2.00 C12AS 4.00 4.00 4.00 4.00 4.00 4.00 Base 0.80 0.80 0.80 0.80 0.80 0.80 Silicate 0.04 0.04 0.04 0.04 0.04 0.04 Perfume 0.35 0.35 0.35 0.35 0.35 0.35 Water and minor ingredients Rest at 100% 2 Protease other than Protease 1 including but not limited to proteases useful in the present invention described herein. **** The pH of the product is about 7 In Examples 12 and 13, any combination of protease enzymes useful in the present invention that are cited herein, inter alia, are replaced by Protease 1 and Protease 2, with substantially similar results. 2. - Dishwashing compositions EXAMPLES 14-19 Compositions for dishwashing Example No. Component 14 15 16 17 18 19 Protease1 0.05 0.50 0.02 0.40 0.10 0.03 Protease2 - - - - 0.40 0.1 TFAA I 0.90 0.90 0.90 0.90 0.90 0.90 AES 12.00 12.00 12.00 12.00 12.00 12.00 Acid 2-methyl 4.50 4.50 - 4.50 4.50 - undecanoic Alcohol C-12 ethoxylated 3.00 3.00 3.00 3.00 3.00 3.00 (4) Amine oxide 3.00 3.00 3.00 3.00 3.00 3.00 Hidrótropo 2.00 2.00 2.00 2.00 2.00 2.00 Ethanol 4.00 4.00 4.00 4.00 4.00 4.00 Mg ++ (as MgCI2) and Mint and minor ingredients **** Rest at 100% 2 Protease other than Protease 1 including but not limited to proteases useful in the present invention described herein. **** The pH of the product is adjusted to 7. In Examples 18 and 19, any combination of the protease enzymes useful in the present invention described herein, inter alia, are replaced by Protease 1 and Protease 2, with substantially results Similar.

EXAMPLE 20 Dishwashing compositions Component A (ADW) B (ADW) C (LDL) STPP 17.5 _ Citrate 15.0 Sodium polyacrylate (MW 4500) 0.80 - Acusol 480N - 5.10 Potassium carbonate 8.30 - Sodium carbonate - 8.50 Silicate 2.1 r K 3.99 - Silicate 2.0r Na 2.00 - Silicate 3.2r Na 5.18 - Aluminum tristearate 0.10 - Nonionic surfactant 2.50 NaAE0.6S 24.70 Glucosamide 3.09 C10E8 4.1 1 Betaine 2.06 Amine oxide 2.06 Magnesium as oxide 0.49 Hydrotrope 4.47 Sodium hypochlorite such as AvCl2 1.15 Protease1 0.01 0.43 0.05 Rest at 100% EXAMPLE 21 Liquid dishwashing compositions (especially suitable under Japanese conditions) Component A B AE1.4S 24.69 24.69 N-cocoyl N-methyl glucamine 3.09 3.09 Amine oxide 2.06 2.06 Betaine 2.06 2.06 Nonionic surfactant 4.1 1 4.1 1 Hydrotrope 4.47 4.47 Magnesium 0.49 0.49 Ethanol 7.2 7.2 Limoneasa 0.45 0.45 Geraniol / BHT - 0.60 / 0.02 Amylase 0.03 0.005 Protease1 0.01 0.43 Rest at 100% EXAMPLE 22 Granulated composition for automatic dishwashing Component B Citrus Acid 15.0 _ Citrate 4.0 29.0 15.0 Acrylate / methacrylate copolymer 6.0 - 6.0 Acrylic acid / acid copolymer - 3.7 maleic Dry carbonate 9.0 - 20.0 Alkali metal silicate 8.5 17.0 9.0 Paraffin - 0.5 Benzotriazole - 0.3 Termamyl 60T 1.6 1 .6 1.6 Protease1 0.2 0.1 0.06 Percarbonate (AvO) 1.5 - Perborate monohydrate - 0.3 1.5 Perborate tetrahydrate - 0.9 Tetraacetylenediamine 3.8 4.4 Diethylenetriaminpenta- 0.13 0.13 0.13 0.13 methylphosphonic acid - 3 times alkylcytoxy-3-ethoxylated acid Nonionic surfactant - 1.5 alkylethoxypropoxy foam suppressant 2.0 - Olin surfactant SLF18 - - 2.0 Sulfate Remainder 100% EXAMPLE 23 Detergent compositions for dishwashing A to F, with high compact density (0.96kg / l) according to the invention: EXAMPLE 24 Granulated detergent compositions for dishwashing, examples A to F with bulk density of 1.02kg / l according to the invention: EXAMPLE 25 A detergent composition is prepared in tablets, examples A to H, according to the present invention, by compressing a granular dishwashing detergent composition at a pressure of 13 KN / cm2 using a standard 12-head rotary press: EXAMPLE 26 Composition in pore tablet for automatic dishwashing Component A (% R.M.) B g R-M.) C q R.M.) Body of the tablet Sodium carbonate 15,348 3,500 5.25 STPP (H2O al12%) 46,482 10,600 9.93 HEDP great. 0.789 0.180 0.28 SKS6 6,578 1,500 2.25 Silicate ratio 2 7,016 1,600 1.65 PB1 10,743 2,450 3.68 Termamyl 2x PCA 0.491 0.1 12 .17 Savinase 0.526 0.120 0.18 Plurafac 3.508 0.800 0.9 BTA 0.263 0.060 0.09 PEG 1.140 0.260 - PEG 4000 - 0.39 Winog 0.439 0.100 0.15 Perfume 0.101 0.023 0.01 Pore filling Citric acid 0.987 0.225 0.23 Bicarbonate 2,600 0.593 0.59 Coloring Sandolan 0.007 0.0017 0.0017 EHRL PEG 400/4000 0.395 0.090 PEG 400 - 0.02 PEG 4000 - 0.08 Amylase 1.412 0.322 0.32 Protease1 0.05 0.268 0.27 3. - Fabric cleaning compositions Granular Fabric Cleaning Composition The granular fabric cleaning compositions of the present invention contain an effective amount of one or more protease enzymes, preferably from about 0.001% to about 10%, most preferably, from about 0.005% to about 5%, most preferably from 0.01% to about 1% by weight of active protease enzyme of the composition. (See patent No. 5,679,630 in the examples).

EXAMPLE 27 Granulated cleaning compositions for fabrics Example No. Component A B C D Protease1 0.10 0.20 0.03 0.05 Protease2 - - 0.2 0.15 Alkylbenzenesulfonate from 22.00 22.00 22.00 22.00 C-I3 linear Phosphate (as 23.00 23.00 23.00 23.00 sodium tripolyphosphates) Sodium carbonate 23.00 23.00 23.00 23.00 Sodium silicate 14.00 14.00 14.00 14.00 Zeolite 8.20 8.20 8.20 8.20 Chelator (acid 0.40 0.40 0.40 0.40 diethylenetriaminpentaacetic acid) Sodium sulphate 5.50 5.50 5.50 5.50 Water Rest at 100% 2 Protease other than Protease 1 including but not limited to proteases useful in the present invention described herein. In Examples 27C and D, any combination of protease enzymes useful in the present invention that are cited herein, inter alia, are replaced by Protease1 and Protease2, with substantially similar results.

EXAMPLE 28 Quenched cleaning composition for fabrics Example No Component A B C D Protease1 0.10 0.20 0.03 0.05 Protease2 - - 0.2 0.1 Alkylbenzenesulfonate 12.00 12.00 12.00 12.00 linear C13 Zeolite A (1-10 26.00 26.00 26.00 26.00 micrometers) (2.3) alkylsulfate 5.00 5.00 5.00 5.00 C-? 2-C .4 secondary Sodium citrate 5.00 5.00 5.00 5.00 Optical brightener 0.10 0.10. 0.10 0.10 Sodium sulphate 17.00. 5:00 PM 5:00 PM Fillers, water, and minor ingredients Rest at 100% 2 Protease other than Protease 1 including but not limited to the additional proteases useful in the present invention described herein. In Examples 28C and D, any combination of the protease enzymes useful in the present invention that are cited herein, inter alia, are replaced by Protease1 and Protease2, with substantially similar results. EXAMPLE 29 Granulated fabric cleaning compositions Components Example No. A B Linear alkyl benzene sulfonate 1 1 .4 10.70 Tallow alkyl sulfate 1.80 2.40 C-Alkylsulfate -? 5 3.00 3.10 Cu-5 alcohol, ethoxylated 7 times 4.00 4.00 Ethoxylated tallow alcohol 1 1 times 1.80 1.80 Dispersant 0.07 0.1 Silicone fluid 0.80 0.80 Trisodium citrate 14.00 15.00 Citric acid 3.00 2.50 Zeolite 32.50 32.10 Copolymer of maleic acid acid 5.00 5.00 acrylic Diethylenetriamine acid 1.00 0.20 pentamethylene phosphonic Protease1 0.1 0.01 Lipase 0.36 0.40 Amylase 0.30 0.30 Sodium silicate 2.00 2.50 Sodium sulfate 3.50 5.20 Polyvinyl pyrrolidone 0.30 0.50 Perborate 0.5 1 Phenol sulfonate 0.1 0.2 Peroxidase 0.1 0.1 Minor ingredients Up to 100 Up to 100 EXAMPLE 30 Granulated fabric cleaning compositions Example No. Component A B Sodium linear alkylene benzene sulphonate 6.5 8.0 Sodium sulfate 15.0 18.0 Zeolite A 26.0 22.0 Sodium Nitrilotriacetate 5.0 5.0 Polyvinylpyrrolidone 0.5 0.7 Tetraacetylethylenediamine 3.0 3.0 Boric acid 4.0 - Perborate 0.5 1 Phenol sulfonate 0.1 0.2 Protease1 0.02 0.05 Fillers up to 100 to 100 (eg: silicates, carbonates, perfumes, water) EXAMPLE 31 Compact granular fabric cleaning compositions Component% by weight Alkylsulfate 8.0 Alkylethoxy Sulfate 2.0 Alcohol mixture of C25 and C45 ethoxylated 3 and 7 times 6.0 Polyhydroxy fatty acid amide 2.5 Zeolite 17.0 Silicate / stratified citrate 16.0 Carbonate 7.0 Acrylic acid maleic acid copolymer 5.0 Discharge polymer 0.4 Carboxymethylcellulose 0.4 Poly (4-vinylpyridine) N-oxide 0.1 Vinylimidazole copolymer and vinylpyrrolidone 0.1 PEG2000 0.2 Protease1 0.03 Lipase 0.2 Cellulase 0.2 Tetraacetylethylenediamine 6.0 Percarbonate 22.0 Ethylenediamindisuccinic acid 0.3 3.5-4'-disodium disulfonate 3,5'-bis (2-morpholin-4-anilin-s-triazine-6-ylamin) stilbene foam buffer 0.25 Disodium-4,4'-bis (sulfoestiril) ) biphenyl 0.05 Water, perfume and minor ingredients up to 100 EXAMPLE 32 Granulated fabric cleaning composition Component | % in weigh Linear alkylbenzenesulfonate 7.6 C6-C Alkylsulfate .8 1.3 Alcohol of C.4..5 ethoxylated 7 times 4.0 Coconut-alkyl-dimethyl hydroxyethylammonium chloride 1.4 Dispersant 0.07 Silicone fluid 0.8 Trisodium Citrate 5.0 Zeolite 4A 15.0 Acrylic acid 4.0 maleic acid copolymer Diethylenetriaminpentamethylenephosphonic acid 0.4 Perborate 15.0 Tetraacetylethylenediamine 5.0 Smectite clay 10.0 Poly (oxyethylene) (MW 300,000) 0.3 Protease1 0.02 Lipase 0.2 Amylase 0.3 Cellulase 0.2 Sodium silicate 3.0 Sodium carbonate 10.0 Carboxymethylcellulose 0.2 0.2 Brighteners Water, perfume and minor ingredients up to 100 EXAMPLE 33 Granulated composition of I impieza of fabrics Component% < in weigh Linear alkylbenzene sulfonate 6.92 Tallow tallow 2.05 Alcohol of C 4.15 ethoxylated 7 times 4.4 C12-.5-ethoxylated alkyl ethoxylate 3 times 0.16 Zeolite 20.2 Citrate 5.5 Carbonate 15.4 Silicate 3.0 Acrylic acid 4.0 maleic acid copolymer Carboxymethyl cellulase 0.31 Dirt releasing polymer 0.30 Protease1 0.1 Lipase 0.36 Cellulase 0.13 Perborate tetrahydrate 1 1.64 Perborate monohydrate 8.7 Tetraacetylethylenediamine 5.0 Diethylenetriaminpentamethylphosphonic acid 0.38 Magnesium sulfate 0.40 Brightener 0.19 Perfume, silicone, foam suppressors 0.85 Ingredients under 100 EXAMPLE 34 Granulated fabric cleaning composition Component A B Base granules LAS / AS / AES (65/35) 9.95 - - LAS / AS / AES (70/30) - 12.05 7.70 Aluminum silicate 14.06 15.74 17.10 Sodium carbonate 1 1.86 12.74 13.07 Sodium silicate 0.58 0.58 0.58 NaPAA solids 2.26 2.26 1.47 PEG solids 1.01 1.12 0.66 Polishers 0.17 0.17 0.1 1 DTPA - - 0.70 Sulfate 5.46 6.64 4.25 Desaerator 0.02 0.02 0.02 Humidity 3.73 3.98 4.33 Minor ingredients 0.31 0.49 0.31 Aerosol B.O.T. Nonionic surfactant 0.50 0.50 0.50 Components of agglomerates LAS / AS (25/75) 11.70 9.60 10.47 Aluminum silicate 13.73 11.26 12.28 Carbonate 8.1 1 6.66 7.26 PEG 4000 0.59 0.48 0.52 Moisture / minor ingredients 4.88 4.00 4.36 Functional Additives Sodium Carbonate 7.37 6.98 7.45 Perborate 1.03 1.03 2.56 Base covering AC - 1.00 - NOBS - - 2.40 Dirt-free polymer 0.41 0.41 0.31 Cellulase 0.33 0.33 0.24 Protease1 0.1 0.05 0.15 Scale AE 0.40 0.40 0.29 Liquid Spray Perfume 0.42 0.42 0.42 Non-ionic aerosol 1.00 1.00 0.50 Ingredients less than 100 EXAMPLE 35 Granular fabric cleaning composition EXAMPLE 36 Granular fabric cleaning compositions EXAMPLE 37 The following granular laundry detergent compositions 37 AC, according to the invention, were prepared, which are of principal utility under washing conditions. in European machines.

EXAMPLE 38 The following formulations are examples of compositions according to the invention, which may be in the form of granules or in the form of a tablet.

EXAMPLE 39 The laundry detergent granular compositions 39 A-E are of primary utility under washing conditions in Japanese machines, and are prepared according to the invention: Liquid fabric cleaning compositions The liquid fabric cleaning compositions of the present invention preferably contain an effective amount of one or more protease enzymes, preferably from about 0.001% to about 10%, most preferably, about 0.001% a about 1%, most preferably still from 0.001% to about 0.1% by weight of active protease enzyme of the composition. (See patent No. 5,679,630 in the examples).

EXAMPLE 40 Liquid fabric cleaning compositions Example No Component A B C D E Protease 0.05 0.03 0.30 0.03 0.10 Protease2 - - - 0.1 0.20 C? 2-C alkylsulfate? , Na 20.00 20.00 20.00 20.00 20.00 2-Butyloctanoic acid 5.00 5.00 5.00 5.00 5.00 5.00 Sodium citrate 1.00 1.00 1.00 1.00 1.00 alcohol ethoxylate Cyan (3) 13.00 13.00 13.00 13.00 13.00 Monoethanolamine 2.50 2.50 2.50 2.50 2.50 2.50 Water / propylene glycol / ethanol ( 100: 1 :: 11)) Remainder at 10% 2 Protease other than Protease 1 including but not limited to the additional proteases useful in the present invention described herein. In Examples 40D and E, any combination of the protease enzymes useful in the present invention that are cited herein, inter alia, are replaced by Protease1 and Protease2, with substantially similar results.

EXAMPLE 41 Liquid compositions for fabric cleaning Example No.

Component A B Alkenyl succinic acid of C .2-14 3.0 8.0 Citric acid monohydrate 10.0 15.0 C? 2- Sodium Alkylsulfate 8.0 8.0 Sodium sulfate alcohol C 12-15 2 times - 3.0 ethoxylated alcohol C 12-15 7 times ethoxylated - 8.0 Diethylenetriaminpenta (methylenephosphonic acid) 0.2 - Oleic acid 1.8 - Ethanol 4.0 4.0 Propanodiol 2.0 2.0 Protease1 0.01 0.02 Polyvinyl pyrrolidone 1.0 2.0 Foam suppressor 0.15 0.15 NaOH up to pH 7.5 Perborate 0.5 1 Phenolsulfonate 0.1 0.2 Peroxidase 0.4 0.1 Water and minor ingredients up to 100% EXAMPLE 42 Lined cloth cleaning compositions Example No.

Component 40 NaLAS (100% am) 16 Neodol 21.5 Citrate 6.8 EDDS 1.2 Dispersant 1.3 Perborate 12 Ester of N-nonanoyl-6- 6-aminocaproic acid phenolsulfonate Protease1 (% pure enzyme) 0.03 Amylase 0.40 Cellulase 0.03 Solvent (BPP) 18.5 Polymer 0.1 Carbonate 10 FWA 15 0.2 TiO2 0.5 PEG 8000 0.4 Perfume 1.0-1 .2 0.06 foam suppressor Water and minor ingredients Up to 100% EXAMPLE 43 Lined cloth cleaning compositions Example No. Component A _B DIH2O 38.63 _.

MEA 0.48 9.0 NaOH 4.40 1.0 Pdiol 4.00 10.0 Citric acid 2.50 2.0 Sodium sulphate 1.75 - DTPA 0.50 1 .0 Premix of FWA (Br 15 / MEA / NI 23-9) 0.15 0.15 Na C25AEI.80S 23.50 - AE3S (H) - 4.0 C1 1.8HLAS 3.00 14.0 Neodol 2.00 6.0 EtOH 0.50 2.0 Formate of Ca * 0.10 0.1 Borax Premix 2.50 - (Borax / MEA / Pdiol / Citric acid) Boric Acid - 1.0 C10 APA 1.50 - TEPA 105 1.20 - FA C12-18 5.00 - Neptune LC 0.50 - Colorant 0.0040 0.0015 Cellulase 0.053 0.2 Amylase 0.15 0.2 Protease1 0.1 0.1 DC-2-3597 0.12 0.2 Rapeseed FA 6.50 4.0 Water and minor ingredients up to 100% EXAMPLE 44 Lined cloth cleaning compositions Component 44 NaOH 5.50 Pdiol 6.90 Citric acid 1.50 DTPA 1.50 Premix FWA (Br 15 / MEA NI 23-9) 0.15 AE3S (H) 2.50 LAS (H) 13.0 Neodol 2.00 EtOH 3.50 Ca Formate * 0.10 Boric Acid 1.00 Clay 4.00 Amylase 0.15 Protease1 0.02 Fatty Acid 16.50 Water and minor ingredients Up to 100% EXAMPLE 45 Fabric Cleaning Liguid Compositions The liquid fabric cleaning composition of particular utility under washing conditions in Japanese machines, is prepared according to the invention: EXAMPLE 46 Lightweight fabric cleaning composition The liquid fabric cleaning composition of particular utility under washing conditions in Japanese machines, as well as washing fine fabrics, is prepared according to the invention: Fabric Cleaning Bar Compositions The fabric cleaning bar compositions of the present invention, suitable for manual washing of soiled fabrics, preferably contain an effective amount of one or more protease enzymes, preferably from about 0.001% to about 10%, most preferably, from about 0.001% to about 1%, most preferably still from 0.01% to about 1% by weight of active protease enzyme of the composition. (See patent No. 5,679,630 in the examples).

EXAMPLE 47 Compositions in cleaning rod for fabrics Example No. Component A B C D Protease1 0.3 0.01 0.02 Protease2 - 0.4 0.1 C12-C16 alkyl sulfate, 20.0 20.0 20.0 20.00 Na N-methyl glucamide from C.2- 5.0 5.0 5.0 5.00 C-? 4 Alkylbenzenesulfonate of 10.0 10.0 10.0 10.00 Cn-Cis, Na sodium pyrophosphate 7.0 7.0 7.0 7.00 Sodium tripolyphosphate 7.0 7.0 7.0 7.00 Zeolite A (0.1 -.10μ 5.0 5.0 5.0 5.00 Carboxymethylcellulose 0.2 0.2 0.2 0.20 Polyacrylate (MW 1400) 0.2 0.2 0.2 0.20 Coconut Monoethanolamide 5.0 5.0 5.0 5.00 Brightener, perfume 0.2 0.2 0.2 0.20 CaSo4 1.0 1 -0 1.0 1.00 MgSO4 1.0 1.0 1.0 1.00 Water 4.0 4.0 4.0 4.00 Filler * rest at 100% * They can be selected from suitable materials such as CaCo3, talc, clay, silicates and the like. Protease other than Protease 1: i. Including but not limited to proteases useful in the present invention described herein. In Examples 47C and D, any combination of protease enzymes useful in the present invention that are cited herein, inter alia, are replaced by Protease1 and Protease2, with substantially similar results. 4. - Oral cleansing compositions Oral cleansing compositions (dentifrices, toothpastes, toothpastes, dental gels, dental powders, mouthwashes, mouth sprays, mouth gels, chewing gum, lozenges, sachets, tablets, biogels, prophylaxis pastes, solutions for dental treatments, and the like) typically contain a pharmaceutically acceptable amount of one or more protease enzymes, preferably from about 0.0001% to about 20%, most preferably from about 0.001% to about 10%, most preferably still around from 0.01% to about 5% by weight of active protease enzymes, useful for removing proteinaceous stains from teeth or dentures. (See also U.S. Patent No. 5,679,630 in the examples).

EXAMPLE 48 Tooth composition Example No.

Component B D Protease1 0.4 0.35 0.15 0.2 Sorbitol (35,000 solution 35,000 35,000 35,000 aqueous 70%) PEG-6 * 1,000 1,000 1,000 1,000 Dental abrasive 20,000 20,000 20,000 20,000 silica ** Sodium fluoride 0.243 0.243 0.243 0.243 Titanium dioxide 0.500 0.500 0.500 0.500 Sodium saccharine 0.286 0.286 0.286 0.286 Sodium alkylsulfate 4.000 4.000 4.000 4.000 (aqueous solution at 27.9%) Flavor 1.040 1.040 1.040 1.040 Polymer of 0.300 0.300 0.300 0.300 carboxivinil *** Carrageenan **** 0.800 0.800 0.800 0.800 Water Rest at 100% * PEG-6 = Polyethylene glycol with a molecular weight of 600. ** Precipitated silica identified as Zeodent 1 19 offered by J.M. Huber. *** Carbopol offered by B.F. Goodrich Chemical Company **** Iota Carrrageenan offered by Hercules Chemical Company.

EXAMPLE 49 Mouthwash composition Example No.

Component \ B C D Protease1 0.3 0.75 0.5 1.00 alcohol SDA 40 8.00 8.00 8.00 8.00 Taste 0.08 0.08 0.08 0.08 Sodium fluoride 0.05 0.05 0.05 0.05 Glycerin 10.00 10.00 10.00 10.00 Sweetener 0.02 0.02 0.02 0.02 Benzoic acid 0.05 0.05 0.05 0.05 Sodium hydroxide.

Coloring 0.04 0.04 0.04 0.04 Water Rest at 100% EXAMPLE 50 Composition in tablet Example No.

Component B D Protease1 0.01 0.03 0.10 0.02 Sorbitol 17.50 17.50 17.50 17.50 Mannitol 17.50 17.50 17.50 17.50 Starch 13.60 13.60 13.60 13.60 Sweetener 1.20 1.20 1.20 1.20 Taste 1 1.70 1 1 .70 1 1.70 1 1.70 Color 0.10 0.10 0.10 0.10 Corn Syrup 100% Rest EXAMPLE 51 Chewing gum composition Example No.

Component A B C D Protease1 0.03 0.02 0.10 0.05 Sorbitol crystals 38.44 38.40 38.40 38.40 Paloja-rubber base T * 20.00 20.00 20.00 20.00 Sorbitol (70% aqueous solution) 22.00 22.00 22.00 22.00 Mannitol 10.00 10.00 10.00 10.00 Glycerin 7.56 7.56 7.56 7.56 Taste 1.00 1.00 1.00 1.00 * Supplied by L.A. Dreyfus Company.

. - Denture Cleaning Compositions Denture cleaning compositions typically contain an effective amount of one or more protease enzymes, preferably from about 0.0001% to about 50%, most preferably from about 0.001% to about 35%, most preferably even about 0.01% to about 20% by weight of active protease enzyme of the composition and a denture cleaning vehicle. (See also U.S. Patent No. 5,679,630 in the examples).

EXAMPLE 52 Two-layer effervescent tablet for cleaning dentures Example No.

Component B C D Acid layer Protease1 1.0 1.5 0.01 0.05 Tartaric acid 24.0 24.0 24.0 24.0 Sodium carbonate 4.0 4.0 4.00 4.00 Sulfamic acid 10.0 10.0 10.00 10.00 PEG 20,000 4.0 4.0 4.00 4.00 Sodium bicarbonate 24.50 24.5 24.50 24.50 Potassium persulfate 15.0 15.0 15.00 15.00 Sodium pyrophosphate acid 7.0 7.0 7.00 7.00 Pyrrogenic silica 2.0 2.0 2.00 2.00 Tetraacetylethylenediamine 7.0 7.0 7.00 7.00 Ricinoleyl sulfosuccinate 0.5 0.5 0.50 0.50 Taste 1.0 1.0 1.00 1.00 Alkaline layer Sodium perborate Monohydrate 32.0 32.0 32.0 32.0 Sodium bicarbonate 19.0 19.0 19.00 19.00 EDTA 3.0 3.0 3.00 3.00 Sodium tripolyphosphate 12.0 12.0 12.00 12.00 PEG 20,000 2.0 2.0 2.0 2.0 Potassium persulfate 26.0 26.0 26.00 26.00 Sodium carbonate 2.0 2.0 2.00 2.00 Pyrrogenic silica 2.0 2.0 2.00 2.00 Coloring / flavor 2.0 2.0 2.00 2.00 Although the particular embodiments of the present invention have been described, it will be obvious to those skilled in the art that various changes and modifications of the present invention can be made without departing from the scope and spirit of the invention itself. Its intention is to cover, in the appended claims, all the determined modifications that are within the scope of the invention. The compositions of the present invention can be prepared in a suitable manner by any method chosen by the formulator, the non-limiting examples of which are described in E.U. 5,691, 297, Nassano et. al, issued November 1, 1997; E.U. 5,574,005 Welch et al., Issued November 12, 1996, E.U. 5,569, 645 Dinniwell et al., Issued October 29, 1996; E.U. 5,565,422 Del Greco et al., Issued October 15, 1996; E.U. 5,516,448 Capeci et al, issued May 14, 1996; E.U. 5,489,392 Capeci et al., Issued February 6, 1996; E.U. 5,486,303 Capeci et al., Issued January 23, 1996, all of which are incorporated herein by reference. In addition to the above examples, the cleaning compositions of the present invention can be formulated into any suitable laundry detergent composition, the non-limiting examples of which are described in E.U. 5,679,630 Baec et al., Issued October 21, 1997; E.U. 5,565,145 Watson et al., Issued Oct. 15, 1996; E.U. 5,478,489 Fredj et al., Issued December 26, 1995; E.U. 5,470,507 Fredj et al., Issued November 28, 1995; E.U. 5,466,802 Panandiker et al., Issued November 14, 1995; E.U. 5,460,752 Fredj et al., Issued October 24, 1995; E.U. 5,458,810 Fredj et al., Issued October 17, 1995; E.U. 5,458,809 Fredj et al., Issued October 17, 1995; E.U. 5,288,431 Huber et al., Issued February 22, 1994, all incorporated herein by reference. As the invention has already been described in detail, with reference to preferred embodiments and examples, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the scope of the invention, which should not be considered. as limited to what is represented in the detailed description.

Claims

NOVELTY OF THE INVENTION CLAIMS

1. - A composition for cleaning fabrics and / or hard surfaces and / or for dishwashing, characterized in that it comprises: (a) an effective amount of a protease variant, wherein said protease variant includes a substitution of a residue of amino acid with another naturally occurring amino acid residue at an amino acid residue position corresponding to position 103 of the subtilisin of Bacillus amyloliquefaciens in combination with a substitution of an amino acid residue with another naturally occurring amino acid residue, in one or more positions of amino acid residues corresponding to positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37 , 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 74, 75, 76, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104 , 106, 107, 109, 1 1 1, 1 14, 1 16, 1 17, 119, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218 , 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257 , 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of Bacillus amyloliquefaciens subtilisin; wherein each protease variant includes a substitution of amino acid residues at the positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the amino acid residue positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and b) one or more auxiliary cleaning materials.

2. The cleaning composition according to claim 1, further characterized in that said protease variant is derived from a subtilisin of Bacillus, preferably subtilisin 309 or subtilisin of Bacillus lentus.

3. The cleaning composition according to claim 1, further characterized in that said protease variant includes substitutions of the amino acid residues in the position 103 and in one or more of the following positions 236 and 245, preferably in the positions 103 and 236, and in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104, 109, 130, 131, 159, 183, 185, 205, 209 , 210, 21 1, 212, 213, 215, 217, 230, 232, 248, 252, 257, 260, 270 and 275; or in positions 103 and 245 and in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104, 109, 130, 131, 159, 170, 183, 185 , 205, 209, 210, 21 1, 212, 213, 215, 217, 222, 230, 232, 248, 252, 257, 260, 261, 270 and 275; and most preferably at positions 103, 236 and 245 and at one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 104, 109, 130, 131, 159, 183 , 185, 205, 209, 210, 21 1, 212, 213, 215, 217, 230, 232, 243, 248, 252, 257, 260, 270 and 275.

4 - The cleaning composition according to claim 1 , further characterized in that said protease variant includes a series of substitutions selected from the group consisting of: 12 102/103/104/159/212 232/236/245/248/252; 12/76/103/104/130/170/185/222/243/245; 12/76/103/104/130/222/245/261; 12/76/103/104/130/222/245; 12/76/103/104/222/245; 61/68/103/104/159/232/236/245/248/252; 62/103/104/159/213/232/236/245/248/252; 62 103/104/109/159/213/232/236/245/248/252; 62/103/104/159/232/236/245/248/252; 62/101/103/104/159/212/213/232/236/245/248/252; 62 103/104/130/159/213/232/236/245/248/252; 68/103/104/159/232/236/245/248/252/270; 68/103/104/159/185/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252; 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/213/232/236/245/248/252; 68/103/104/159/230/232/236/245; 68/76/103/104/159/209/232/236/245; 68/103/104/232/236/245/248/257/275; 68/103/104/213/232/236/245 248/252; 68/103/104/159/232/236/245/248/252; 68/103/104/159/209/232/236/245; 68/76/103/104/159/236; 68/76/103/104/159/236/245; 68/76/103/104/159/232/236/245; 68/103/104/159/232/236/245/252; 68/103/104/159/232/236/245; 68/103/104/159/232/236/245/257; 68/76/103/104/159/211/232/236/245; 68/76/103/104/159/215/232/236/245; 68/103/104/159/210/232/236/245; 68/103/104/159/213/232/236/245/260; 68/76/103/104/159/213/232/236/245/260; 68/103/104/159/236; 68/76/103/104/159/210/232/236/245/260; 68/103/104/159/236/245; 68/103/104/159/183/232/236/245/248/252; 68/76/103/104/159/236/245; 68/103/104/232/236/245/257/275; 68/103/104/159/213/232/236/245; 76/103/222/245: 76/103/104/222/245; 76/103/104/159/232/236/245; 76/103/104/159/213/232/236/245/260; 76/103/104/159; 76/103/104/131/159/232/236/245/248/252; 97/103/104/159/232/236/245/248/252; 98/102/103 / 104/159/212/232/236/245/248/252; 98/103/104/159/232/236/245/248/252: 101/103/104/159/232/236/245/248/252; 102/103/104/159/232/236/245/248/252; 103/104/159/232/236/245; 103/104/159/232/236/245/248/252; 103/104 / 159/205/209/232/236/245/257 103/104/159/232/245/248/252; 103/104/159/205/209/210/232/236/245/257; 103/104/159/213/232/236/245/248/252; 103/104/159/217/232/236/245/248/252; 103/104/130/159/232/236/245/248/252; 103/104/159/230/236/245; 103/104/159/236/245; 103/104/159/248/252/270; 103/104/13 1/159/232/236/245/248/252; 103/104/159/205/209/232/236/245; and 103/104/159/232/236/245/257.

5. - The cleaning composition according to claim 4, further characterized in that said protease variant includes a series of substitutions selected from the group consisting of: 12R / 76D / 103A / 104T / 130T / 222S / 245R; 12R / 76D / 103 A / 104I / 222S 245R; 12R / 102.V103A / 104I / 159D / 2I2G / 232V / 236H / 245R 248D / 2S2K; 12R / 76D / 103 A / 104T / 130G / 222S / 245R / 261 D; 12R / 76D / 103A / 104T / 130G / 170S / 185D / 222S / 243D / 245R; 61 E / 68A 103.A / 1041 / 159D / 232V / 236H / 245R 248D / 2S2K; 62D / 103A / 104I / 109R / I59D / 213R 232V / 236H / 245R / 248D / 252K; 62D / 103A / 104I / 159D / 213R 232V / 236H / 245R 248D / 252K; 62D / 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; Ó2D / 103A / 104I / 130G / 159D / 213R 232V / 236H / 24SR 248D / 252K; 62D / 101G / 103A / 104I / 159D / 212G / 213R / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252K / 270A; 68A 76D / 103 A / 1041 / 159D / 213R / 232V / 236H / 245R / 260A; 68A / 103A / 104I / 159D / 236H; 68A / 103A / 104I / 159D / 236H / 245R; 68A 76D / 103A / 104I / 159D / 210I / 232V / 236H / 245R / 260A; 68A / 103 A 1041 / 159D / 183D / 232V / 236H / 245R / 248D / 252K; 68 A / 103 A / 1041 / 159D / 209W / 232V / 236H / 245R; 68A / 76D / 103 A / 1041 / 159D / 21 1 R / 232V / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 215R 232V / 236H / 245R; 68A / 103A 104I / 159D / 2I3R / 232V / 236H / 245R / 260A; 68A / 76D / 103A / 104I / 159D / 236H: 68A 76D / 103A 1041 / 159D / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 232V / 236H / 245R; 68A 103 A 1041 / 159D / 232V / 236H / 245R / 252K; 68A / 103 A 1041 / 159D / 232V / 236H / 245R: 68A 103A / 104I / 159D / 232V / 236H / 245R / 257V; 68A 103A / 104I / 159D / 185D / 232V / 236H / 245R / 248D / 252; 68A / 103A 1041 / 159D / 210L / 232V / 236H / 245R / 248D / 252K; A / 103A / 1041 / 159D / 185D / 210L / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 213E / 232V / 236H / 245R / 248D / 252; 68A 103 A / 1041 / 159D / 230V / 232V / 236H / 245R; 68A / 76D / 103A / 1041 / 159D / 209 / 232V / 236H / 245R; 68A / 103A / 104I / 232V / 236H / 245R 248D / 257V / 275H; 68A 103A / 1041 / 232V / 236H 245R 257V / 275H; 68A / 103 A / 1041 / 213E / 232V / 236H / 245R / 248D / 252K; 68A / 103 to 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 68A 103 A / 1041 / 159D / 210I / 232V / 236H / 245R; 68A / 103A 104I / 159D / 210L / 232V / 236H / 245R; 68A / 103 A / 1041 / 159D / 213G / 232V / 236H / 245R; 76D / 103A 222S / 245R; 76D / 103A / 104I / 222S / 245R; 76D / 103A / 104I / 159D / 232V / 236H / 245R; 76D / 103A / 104I / 159D; 76D / 103A / 104I / 131V / 159D / 232V / 236H / 245R / 248D / 252K; 76D / 103 A / 1041/1 S9D / 213R / 232V / 236H / 245R / 260A; 97E / 103 A 1041 / 159D / 232V / 236H / 245R / 248D / 252; 98L / 103A 104I / 159D / 232V / 236H / 245R 248D / 252K; L / 102A / 103A / 104I / 159D / 212G / 232V / 236H / 245R / 248D / 252K; 101G / 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252; 102A 103A 104I / 159D / 232V / 236H / 245R / 248D / 252K; 103A 1041 / 159D / 232V / 236H 245R / 248D / 252K; 10 A 104 I / 159 D / 213 R 232 V / 236 H / 245 R / 248 D / 252 K; 103A / 1041 / 130G / 159D / 232V / 236H / 245R 248D / 252K; 103 A / 1041 / 159D / 230V / 236H / 245R; 103A / 104I / 159D / 217E / 232V / 236H / 245R / 248D / 252; 103 A / 1041 / 159D / 236H / 245R; 103 A l 041 / 159D / 248D / 252K / 270V; 103 A / 1041 / 159D / 232V / 236H / 245R; 103A / 104I / 159D / 205I / 209W / 232V / 236H7245R; 103A / 104I / 159D / 232V / 236H / 245R / 257V: 103 A / 1041 / 159D / 205I / 209W / 232V / 236H 245R / 257V; 103A / 104I / 131 V / 159D / 232V / 236H / 245R 248D / 252K; 103A 104I / 159D / 205I / 209W / 210I / 232V / 236H / 245R / 257V; ar.d 103.A / 104I / 159D / 232V / 245R / 248D / 252K.

6. - The cleaning composition according to claim 1, further characterized in that said cleaning adjunct materials are selected from the group consisting of surfactants, solvents, pH regulators, enzymes, agents for the release of dirt, agents for the removal of clay soils, dispersing agents, brighteners, suds suppressors, fabric softeners, foam enhancers, enzyme stabilizers, detergency builders, other bleaching agents, dyes, perfumes, chelating agents, and mixtures thereof.

7. The cleaning composition according to claim 6, further characterized in that said cleaning adjunct materials comprise at least one detersive surfactant, preferably a branched surfactant, more preferably a branched surfactant in the middle region of its chain.

8. The cleaning composition according to claim 7, further characterized in that the cleaning adjunct materials comprise at least about 0.1% by weight of surfactant of the composition, said surfactant comprising materials selected from the group consisting of alkylbenzene sulfonates. , primary alkyl sulphates, secondary alkyl sulphates, alkyl alkoxy sulfates, alkyl alkoxy carboxylates, alkyl polyglucosides and their corresponding sulphated polyglucosides, alpha-suifonated fatty acid esters, alkyl and alkyl phenol alkoxylates, betaines and sulfobetaines, amine oxides, N-methyl glucamides, alcohol primary non-ionic ethoxylates, ethoxy / non-ionic propoxy mixed with primary alcohol, and mixtures thereof.

9. The cleaning composition according to claim 8, further characterized in that it comprises at least about 5% detergency builder selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof.

10. The cleaning composition according to claim 6, further characterized in that said cleaning adjunct materials comprise at least one detersive enzyme selected from the group consisting of cellulases, lipases, amylases, phospholipases, other proteases, peroxidases, and mixtures from the same.

11. The cleaning composition according to claim 6, further characterized in that said cleaning adjunct materials comprise at least one bleaching agent selected preferably from the group consisting of percarbonates, perborates and mixtures thereof, and optionally comprising further at least one bleach activator selected from the group consisting of benzoyloxybenzenesulfonate (BOBS), nonanoyloxybenzenesulfonate (NOBS), decanoyloxybenzenesulfonate (CBS OBS), octanoyloxybenzenesulfonate (C8 OBS), perhydrolyzable esters, sodium salt of 4- [ N- (nonaoyl) amino hexanoyloxy-benzenesulfonate (NACA-OBS), lauryloxybenzenesulfonate (LOBS or C 2 OBS), 10-undecenoyloxybenzenesulfonate (UDOBS or Cu OBS with unsaturation at position 10) and decanoyloxybenzoic acid (DOBA), and mixtures thereof, and optionally further comprising at least one bleaching catalyst, preferably 3- (3,4-dihydroisoquinolium) prop ansulfonate.

12. The cleaning composition according to claim 1, further characterized in that said cleaning composition is a fabric cleaning composition, preferably in the form of a liquid, granule, bar, tablet, gel, powder or foam, comprising at least about 5% surfactant and therefore less about 5% detergency builder by weight of the composition.

13. The cleaning composition according to claim 1, further characterized in that said cleaning composition is a fabric cleaning composition, comprising: (a) from about 0.0001% to about 10% by weight of said protease variant; (b) at least about 5% by weight of a surfactant preferably selected from the group consisting of alkybenzenesulfonates, primary alkyl sulphates, secondary alkyl sulphates, alkyl alkoxy sulfates, alkyl alkoxy carboxylates, alkyl polyglucosides and their corresponding sulphated polyglycosides, esters of alpha-sulfonated fatty acid, alkyl and alkyl phenol alkoxylates, betaines and sulfobetaines, amine oxides, N-methyl glucamides, primary alcohol non-ionic ethoxylates, ethoxy / non-ionic propoxy mixed with primary alcohol, and mixtures thereof; and wherein further the detergency builder is selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof; and (c) at least about 5% by weight of a builder preferably selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof.

14. The cleaning composition according to claim 25, further characterized in that it is in the form of a concentrated granular composition for cleaning fabrics, comprising at least about 15% surfactant.

15. A method for cleaning fabrics, characterized in that said method comprises contacting a fabric that needs to be cleaned, with a cleaning composition according to claims 12 or 13.

16.- The cleaning composition in accordance with claim 1, further characterized in that said cleaning composition is a dishwashing composition, preferably in the form of a liquid, granule, powder, gel or tablet, comprising: (a) from about 0.0001% to about 10% by weight of said protease variant; and (b) from about 0.1% to about 10% by weight of a surfactant.

17. A method for cleaning dishes, said method comprising contacting a dish that needs to be cleaned, with a cleaning composition according to claim 16.

18. A composition for personal cleaning, characterized in that it comprises: ( a) an effective amount of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another naturally occurring amino acid residue at an amino acid residue position corresponding to position 103 of the subtilisin of Bacillus amyloliquefaciens in combination with a substitution of an amino acid residue with another naturally occurring amino acid residue, in one or more positions of amino acid residues corresponding to the positions: 1, 3, 4, 8, 9, 10, 12 , 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76 , 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104 , 106, 107, 109, 1 1 1, 1 14, 1 16, 1 17, 1 19, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147 , 158, 159, 160, 166, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of subtilisin Bacillus amyloliquefaciens; wherein said protease variant includes a substitution of amino acid residues at the positions corresponding to positions 103 and 76, where there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the residue positions of amino acids corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and b) one or more auxiliary cleaning materials.

19. The composition for personal cleansing according to claim 18, further characterized in that said composition for personal cleansing comprises: (a) from about 0.001% to about 5%, preferably from 0.001% to about 2%, more preferably from about 0.002% to about 0.8% by weight of said protease variant; and (b) from about 0.1% to about 95% by weight of a surfactant system preferably comprising a surfactant selected from the group consisting of anionic carboxylates, amine oxides, alkyl glucosides, glucosamides, alkyl sulfates, alkyl ether sulphates, acyl isethionates, alkyl sulfosuccinates, alkyl phosphate esters, ethoxylated phosphate esters, alkyl glyceryl ether sulfonates, and mixtures thereof, more preferably comprising a surfactant selected from the group consisting of soaps, acyl glutamates, alkyl sarcosinates, lauramine oxides, cocoamine oxides, cocoamidopropylamine oxides, decyl glucosides, lauryl sulfates, laureth sulfates, C 2-18 acyl isethionates, and mixtures thereof; and (c) optionally, from about 0.05% to about 50% by weight of an enzyme stabilizer.

20. The composition for personal cleansing according to claim 19, further characterized in that said surfactant is soap at a level of at least about 2%, preferably at least about 10%, more preferably at least about 25% by weight of the cleaning composition.

21. The composition for personal cleansing according to claim 20, further characterized in that the ratio of soap: protease variant is from about 2,000: 1 to about 8: 1, preferably from about 400: 1 to about 40: 1.

22. A method for personal cleansing, characterized in that said method comprises contacting a part of the body of a human or lower animal that needs to be cleaned, with a cleaning composition according to claim 18.

23.- A composition for cleaning fabrics and / or hard surfaces and / or for washing dishes, characterized in that it comprises: (a) an effective amount of a protease variant, wherein said protease variant includes a substitution of one amino acid residue with another naturally occurring amino acid residue in one or more positions of amino acid residues corresponding to positions 230 and 232 of the subtilisin of Bacillus amyloliquefaciens; and (b) one or more cleaning attachment materials.

24. The cleaning composition according to claim 23, further characterized in that said protease variant is derived from a subtilisin of Bacillus, preferably subtilisin 309 or subtilisin of Bacillus lentus.

25. The cleaning composition according to claim 23, further characterized in that said protease variant includes substitutions of the amino acid residues in one or more of the following positions selected from the group consisting of: 1) position 230 and in one or more of the following positions: 68, 103, 104, 159, 232, 236 and 245; 2) position 232 and in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183,185, 205, 209 , 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 3) the position 232 and in one or more of the following positions: 103, 104, 236 and 245; 4) the positions 232 and 103 in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 5) the positions 232 and 104 in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 6) the positions 232 and 236 in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 7) the positions 232 and 245 in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 8) the positions 232, 103, 104, 236 and 245 in one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183,185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275;

26. The cleaning composition according to claim 23, further characterized in that said protease variant includes a series of substitutions selected from the group consisting of: 12/102/103/104/159/212/232/236/245/248/252; 61/68/103/104/159/232/236/245/248/252; 62/103/104/130/159/213/232/236/245/248/252; 62/103/104/159/213/232/236/245/248/252; 62/103/104/109/159/213/232/236/245/248/252; 62/103/104/159/232/236/245/248/252; 62/101/103/104/159/212/213/232/236/245/248/252; 68/103/104/159/232/236/245/248/252/270; 68/103/104 / 159/185/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252; 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/213/232/236/245/248 252; 68/103/104/159/230/232/236/245; 68/76/103/104/159/209/232/236/245; 68/103/104/232/236/245/248/257/275; 68/103/104/213/232 236/245/248/252; 68/103/104/159/232/236/245/248/252; 68/103/104/159/209/232/236/245; 68/76/103/104/159/232/236/245; 68/103/104/159/232/236/245/252; 68/103/104/159/232/236/245; 68/103/104/159 / 23Z / 236/245/257; 68/76 / 10J / 104/159/21 1/232/236/245; 68/76/103/104/159/215/232/236/245; 68/103/104/159/210/232/236/245; 68/103/104/159/213/232/236/245/260; 68/76/103 / 104/159/213/232/236/245/260; 68/76/103/104/159 210/232/236/245/260; 68/103/104/159/183/232/236/245/248/252; 68/103/104/232/236/245/257/275; 68/103/104/159/213/232/236/245; 76/103/104/159/232/236/245; 76/103/104/159/213/232/236/245/260; 76/103/104/131/159/232/236/245/248/252; 97/103/104/159/232/236/245/248/252; 98/103/104/159/232/236/245/248/252; 98/102/103/104/159/212/232/236/245/248/252; 101/103/104/159/232/236/245/248/252; 102/103/104 / 159/232/236/245/248/252; 103/104/159/232/236/245, 103/104 / 159/248/252/270; 103/104/159/232/236/245/2 8/25; 103/104/159/205/209/232/236/245/257 103/104/159/232/245/248/252; 103/104/159/205/209/210/232/236/245/257; 103/104/159/213/232/236/245/248/252; 103/104/159/217/232/236/245/248/252; 103/104/130/159/232/236/245/248/252; 103/104/131/159/232/236/245/248/252; 103/104/159/205/209/232/236/245; and 103/104/159/232/236/245/257.

27. - The cleaning composition according to claim 26, further characterized in that said protease variant includes a series of substitutions selected from the group consisting of: 12R 102.V103A / I 04I / 159D / 212G / 232V / 236H / 24SR / 248D / 252K; 61 E 68A / 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 62D / 103A / 1041 / 109R / 159D / 213R / 232V 236H / 245R / 248D / 252K, 62D / Í03A 1041 / 159D / 213R / 232V / 236H / 245R / 248D / 252; 62D / 103A / 104r / 159D / 232V / 236H / 245R / 248D / 252K; 62D / 103A / 104I / 13OG / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 62D / 101G / 103A / 1041 / 159D / 212G / 213R / 232V / 236H / 245R / 248D / 252K; 68A / 76D / 103A / 104I / 159D / 213R / 232V / 236H / 245R 260A; 68A / 76D / 103A / 104I / I 59D / 2101 / 232V / 236H 245R / 260A; 68A / 103 A 1041 / 159D / 183D / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 209W / 232V / 236H / 245R; 68A 76D / 103A / 1041 / 159D / 21 1 R / 232V / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 215R / 232V / 236H / 245R; 68A / 103A / 1041 / 159D / 213R / 232V / 236H / 245R / 260A; 68A / 76D / 103A 1041 / 159D / 232V / 236H 245R; 68A / 103 A / 1041 / 159D / 232V / 236H / 24SR / 252K, 68A / 103A / 1041 / 159D / 232V / 236H / 245R; 68A / 103A / 1041 / 159D / 232V / 236H / 245R / 257V; 68A / 103A / 104I / 159D / 185D / 232V / 236H / 245R 248D / 252K; 68A / 103 A / 1041 / 159D / 210L 232V / 236H 245R / 248D 252 68A / 103 1041 / 159D / 185D / 210L / 232V / 236H / 24SR / 248D / 252K; 68A / 103A / 104I / 159D / 213E / 232V / 236H / 245R / 248D / 252K; 68A / 103 A / 1041 / 159D / 230V / 232V 236H / 245R; 68A / 76D / 103A / 1041 / 159D / 209W / 232V / 236H / 245R; 68A 103A 104I / 232V / 236H / 245R / 248D / 257V 275H; 68A / 103A / 104I / 232V / 236H / 245R / 257V / 275H; 68A 103A / 1041 / 213E / 232V / 236H245R248D / 252K; 68A / 103 A / 1041 / 159D / 232V / 236H / 245R / 248D / 252; 68A 103 A 1041 / 159D / 210I / 232V / 236H245R; 68A / 103A 1041 / 159D / 210L / 232V / 236H245R; 68A 103A / 104I / 159D / 213G / 232V / 236H245R; 68A / 103 A / 1041 / 159D / 232V / 236H245R / 248D / 252K / 270A; 76D / 103 A / 1041 / 159D / 232V / 236H / 245R; 76D / 103A / 1041/131 V / l 59D / 232V / 236H / 245R / 248D / 252K; 76D / 103 A / 1041 / 159D / 213R / 232 V / 236H / 245R260 A; 97E / 103A 104? / 159D / 232V / 236H245R248D / 252K; 98L / 103A 104I / 159D / 232V / 236H / 245R248D / 252K; 98L / 102A / 103A 104I / 159D / 212G / 232V / 236H / 245R248D / 252K; 101G / 103A / 104I / 159D / 232V / 236H / 245R248D / 252K; 102A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252; 103A 104I / 159D / 232V / 236H245R248D / 252K; 103A / 104I / 159D / 2I3R232V / 236H245R / 248D / 252K; 103A / 104I / 130G / 159D / 232V / 236H / 245R248D / 252K; 103A 1041 / 159D / 217E232V / 236H / 245R248D / 252K; 103 A 1041 / 159D / 248D / 252K / 270V; 103A / 104I / 159D / 232V / 236H / 245R; 103A / 104I / 159D / 205I / 209W / 232V / 236H / 245R; 103 A / 1041 / 159D / 232V / 236H / 245R257V; 103A / 104I / 159D / 205I / 209W / 232V / 236H / 245R / 257V; 103A / 104I / 131V / 159D / 232V / 236H / 245R / 248D / 252K; 103A / 1041 / 159D / 205I / 209W / 210? / 232V / 236H / 245R / 257V: and 103A / 1041 / 159D / 232V / 245R / 248D / 252K.

28. - The cleaning composition according to claim 23, further characterized in that said cleaning adjunct materials are selected from the group consisting of surfactants, solvents, pH regulators, enzymes, agents for the release of dirt, agents for the removal of clay soils, dispersing agents, brighteners, suds suppressors, fabric softeners, foam enhancers, enzyme stabilizers, detergency builders, other bleaching agents, dyes, perfumes, chelating agents, and mixtures thereof.

29. The cleaning composition according to claim 28, further characterized in that said cleaning adjunct materials comprise at least one detersive surfactant, preferably a branched surfactant, more preferably a branched surfactant in the middle region of its chain.

30. The cleaning composition according to claim 28, further characterized in that the cleaning adjunct materials comprise at least about 0.1% by weight of the surfactant of the composition, said surfactant comprising materials selected from the group consisting of alkylbenzene sulfonates. , primary alkylsulphates, secondary alkyl sulphates, alkyl alkoxy sulphates, alkyl alkoxy carboxylates, alkyl polyglycosides and their corresponding sulphated polyglycosides, alpha-sulfonated fatty acid esters, alkyl and alkyl phenol alkoxylates, betaines and suiphobetaines, amine oxides, N-methyl glucamides, alcohol primary non-ionic ethoxylates, ethoxy / non-ionic propoxy mixed with primary alcohol, and mixtures thereof.

31. The cleaning composition according to claim 30, further characterized in that it comprises at least about 5% detergency builder selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof.

32. The cleaning composition according to claim 28, further characterized in that said cleaning adjunct materials comprise at least one detersive enzyme selected from the group consisting of cellulases, lipases, amylases, phospholipases, other proteases, peroxidases, and mixtures from the same.

33. The cleaning composition according to claim 28, further characterized in that said cleaning adjunct materials comprise at least one bleaching agent selected preferably from the group consisting of percarbonates, perborates and mixtures thereof, and optionally comprising further at least one bleach activator selected from the group consisting of benzoyloxybenzenesulfonate (BOBS), nonanoyloxybenzenesulfonate (NOBS), decanoyloxybenzenesulfonate (CBS OBS), octanoyloxybenzenesulfonate (C8 OBS), perhydrolyzable esters, sodium salt of 4- [ N- (nonaoyl) amino hexanoyloxy-benzenesulfonate (NACA-OBS), lauroyloxybenzenesulfonate (LOBS or C 2 OBS), 10-undecenoyloxybenzenesulfonate (UDOBS or Cu OBS with unsaturation at position 10) and decanoyloxybenzoic acid (DOBA), and mixtures thereof, and optionally further comprising at least one bleaching catalyst, preferably 3- (3,4-dihydroisoquinolium) prop ansulfonate.

34. The cleaning composition according to claim 23, further characterized in that said cleaning composition is a fabric cleaning composition, preferably in the form of a liquid, granule, bar, tablet, gel, powder or foam, which it comprises at least about 5% surfactant and at least about 5% detergency builder by weight of the composition.

35.- The cleaning composition according to claim 23, further characterized in that said cleaning composition is a fabric cleaning composition, comprising: (a) from about 0.0001% to about 10% by weight of said protease variant; (b) at least about 5% by weight of a surfactant preferably selected from the group consisting of alkylbenzene sulfonates, primary alkyl sulphates, secondary alkyl sulphates, alkyl alkoxy sulfates, alkyl alkoxy carboxylates, alkyl polyglucosides and their corresponding sulphated polyglycosides, esters of alpha-sulfonated fatty acid, alkyl and alkyl phenol alkoxylates, betaines and sulfobetaines, amine oxides, N-methyl glucamides, primary alcohol non-ionic ethoxylates, ethoxy / non-ionic propoxy mixed with primary alcohol, and mixtures thereof; and wherein further the detergency builder is selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof; and (c) at least about 5% by weight of a builder preferably selected from the group consisting of zeolites, polycarboxylates, layered silicates, phosphates, and mixtures thereof.

36.- The cleaning composition according to claim 35, further characterized in that it is in the form of a concentrated granular composition for cleaning fabrics, comprising at least about 15% surfactant.

37.- A method for cleaning fabrics, characterized in that said method comprises contacting a fabric that needs to be cleaned, with a cleaning composition in accordance with claims 34 or 35.

38.- The cleaning composition in accordance with Claim 23, further characterized in that said cleaning composition is a dishwashing composition, preferably in the form of a liquid, granule, powder, gel or tablet, comprising: (a) from about 0.0001% to about 10% by weight of said protease variant; and (b) from about 0.1% to about 10% by weight of a surfactant.

39.- A method for cleaning dishes, said method comprising contacting a dish that needs to be cleaned, with a cleaning composition according to claim 38.

40. - A composition for personal cleansing, characterized in that it comprises: an effective amount of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another naturally occurring amino acid residue, in one or more residue positions of amino acids corresponding to positions 230 and 232 of the subtilin of Bacillus amyloliquefaciens; and (b) one or more cleaning attachment materials.

41. The composition for personal cleansing according to claim 40, further characterized in that said composition for personal cleansing comprises: (a) from about 0.001% to about 5%, preferably from 0.001% to about 2%, more preferably from about 0.002% to about 0.8% by weight of said protease variant; and (b) from about 0.1% to about 95% by weight of a surfactant system preferably comprising a surfactant selected from the group consisting of anionic carboxylates, amine oxides, alkyl glucosides, glucosamides, alkyl sulfates, alkyl ether sulphates, acyl isethionates, alkyl sulfosuccinates, alkyl phosphate esters, ethoxylated phosphate esters, alkyl glyceryl ether sulfonates, and mixtures thereof, more preferably comprising a surfactant selected from the group consisting of soaps, acyl glutamates, alkyl sarcosinates, lauramine oxides, cocoamine oxides, cocoamidopropylamine oxides, decyl glucosides, lauryl sulfates, laureth sulfates, C? 2-? 8 acyl isethionates, and mixtures thereof; and (c) optionally, from about 0.05% to about 50% by weight of an enzyme stabilizer.

42.- The composition for personal cleansing according to claim 41, further characterized in that said surfactant is soap at a level of at least about 2%, preferably at least about 10%, more preferably at least about 25%. % by weight of the cleaning composition.

43.- The composition for personal cleansing according to claim 42, further characterized in that the ratio of soap: protease variant is from about 2,000: 1 to about 8: 1, preferably from about 400: 1 to about 40: 1. .

44.- A method for personal cleansing, characterized in that said method comprises contacting a part of the body of a human or lower animal that needs to be cleaned, with a cleaning composition according to claim 40.

45.- A method for the pretreatment of a fabric that needs to be cleaned, said method comprising contacting said fabric before washing it, with an aqueous solution containing a surfactant with a cleaning composition according to claims 12 or 13.

46.- A method for pretreating a fabric that needs to be cleaned, said method comprising contacting said fabric before washing with an aqueous solution containing a surfactant with a cleaning composition according to claim 34 or 35.

47. - A composition for cleaning fabrics and / or hard surfaces and / or for washing dishes, characterized in that they also include of: (a) an effective amount of a protease variant, wherein said protease variant includes a substitution of an amino acid residue with another naturally occurring amino acid residue at one or more amino acid residue positions corresponding to the positions 62, 212, 230, 232, 252 and 257 of the subtilisin of Bacillus amyloliquefaciens; wherein said protease variant includes substitutions of the amino acid residues in one or more of the following positions selected from the group consisting of: 1) a protease variant including substitutions of amino acid residues at position 62 and one or more than the following positions: 103, 104, 109, 159, 213, 232, 236, 245, 248 and 252; 2) a protease variant including substitutions of the amino acid residues at position 212 and one or more of the following positions: 12, 98, 102, 103, 104, 159, 232, 236, 245, 248 and 252; 3) a protease variant including substitutions of the amino acid residues at position 230 and one or more of the following positions: 68, 103, 104, 159, 232, 236 and 245; 4) a protease variant including substitutions of the amino acid residues at position 232 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 5) a protease variant including substitutions of amino acid residues at position 232 and one or more of the following positions: 103, 104, 236 and 245; 6) a protease variant including substitutions of amino acid residues at positions 232 and 103 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 7) a protease variant including substitutions of the amino acid residues at positions 232 and 104 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 8) a protease variant including substitutions of the amino acid residues at position 232 and 236 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 9) a protease variant including substitutions of the amino acid residues at position 232 and 245 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270 and 275; 10) a protease variant including substitutions of the amino acid residues at position 232, 103, 104, 236 and 245 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 205, 209, 210, 212, 213, 217, 230, 236, 245, 248, 252, 257, 260, 270, and 275; 1 1) a protease variant including substitutions of the amino acid residues at position 252 and one or more of the following positions: 12, 61, 62, 68, 76, 97, 98, 101, 102, 103, 104, 109 , 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248, 270; 12) a protease variant including substitutions of the amino acid residues at position 252 and one or more of the following positions: 103, 104, 236 and 245; 13) a protease variant including substitutions of the amino acid residues at position 252 and 103 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248, and 270; 14) a protease variant including substitutions of the amino acid residues at position 252 and 104 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 15) a protease variant including substitutions of the amino acid residues at position 252 and 236 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 16) a protease variant including substitutions of the amino acid residues at position 252 and 245 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 17) a protease variant including substitutions of amino acid residues at position 252, 103, 104, 236 and 245 and one or more of the following positions: 12, 61, 62, 68, 97, 98, 101, 102, 103, 104, 109, 130, 131, 159, 183, 185, 210, 212, 213, 217, 232, 236, 245, 248 and 270; 18) a protease variant including substitutions of amino acid residues at position 257 and one or more of the following positions: 68, 103, 104, 205, 209, 210, 232, 236, 245 and 275.

48.- One composition for cleaning fabrics and / or hard surfaces and / or for washing dishes, further characterized in that it comprises: (a) an effective amount of a protease variant, wherein said protease variant includes a substitution of a residue of amino acid with another naturally occurring amino acid residue in one or more positions of amino acid residues corresponding to positions 62, 212, 230, 232, 252 and 257 of the subtilisin of Bacillus amyloliquefaciens; wherein said protease variant includes a series of substitutions selected from the group consisting of: 12/102/103/104/159/212/232/236/245/248/252; 61/68/103/104/159/232/236/245/248/252; 62/103/104/130/159/213/232 236/245/248/252; 62/103/104/159/213/232/236/245/248/252, 62/103/104/109/159/213/232/236/245/248/252; 62/103/104/159/232/236/245/248/252; 62/101/103/104/159/212/213/232/236/245/248/252; 68/103/104/159/232/236/245/248/252/270; 68/103/104/159/185/232/236/245/248/252; 68/103/104/159/210/232/236/245/248/252; 68/103/104/159/185/210/232/236/245/248/252; 68/103/104/159/213/232/236/245/248/252: 68/103/104/159/230/232/236/245; 68/76/103/104/159/209/232/236/245; 68/103/104/232/236/245/248/257/275; 68/103/104/213/232/236/245/248/252; 68/103/104/159/232/236/245/248/252; 68/103/104/159/209/232/236/245; 68/76/103/104/159/232/236/245; 68/103/104/159/232/236/245/252; 68/103/104/159/232/236/245; 68/103/104/159/232/236/245/257; 68/76/1037! 04/159/21 1/232/236/245; 68/76/103/104/159/215/232/236/245; 68/103/104/159/210/232/236/245; 68/103/104 / 159/213/232/236/245/260; 68/76/103/104/159/213/232/236/245/260; 68/76/103/104/159/210/232/236/245/260; 68/103/104/159/183/232/236/245/248/252; 68/103/104/232/236/245/257/275; 68/103/104/159/213/232/236/245; 76/103/104/159/232/236/245; 76/103/104/159/213/232/236/245/260; 76/103/104/131/159/232/236/245/248/252; 97/103/104/159/232/236/245/248/252; 98/103/104/159/232/236/245/248/252; 98/102/103/104/159/212/232/236/245/248/252; 101/103/104/159/232/236/245/248/252; 102 / 103/104/159/232/236/245/248/252; 103/104/159/232/236/245: 103/104 / 159/248/252/270; 103/104/159/232/236/245/248/252; 103/104/159/205/209/232/236/245/257 103/104/159/232/245/248/252; 103/104/159/205/209/210/232/236/245/257; 103/104/159/213/232/236/245/248/252; 103/104/1 59/217/232/236/245/248/252; 103/104/130/159/232/236/245/248/252; 103/104/13 1/159/232/236/245/248/252; 103/104/159/205/209/232/236/245; and 103/104 / 159/232/236/245/257.

49. - The cleaning composition according to claim 48, further characterized in that said protease variant includes a series of substitutions selected from the group consisting of: 12R 102A / 103 A / 1041 / 159D / 212G / 232 V / 236H / 24SR 248D / 252K; 6 I / 68A / 103 A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 62D / 103A / 104I / 109R 159D / 213R 232V / 236H 245R / 248D / 252K; 62D / 103 A / 1041 / 159D / 213R 232V / 236H / 245R / 248D / 252K; 62D / 103A / 1041 / 159D / 232V / 236H 245R 248D / 252; 62D / 103A 104I / 130G / 159D / 213R / 232V / 236H / 245R / 248D / 252K; 62D / 101G / 103A / 104I / 159D / 212G / 213R / 232V 236H / 245R / 248D / 252; 68A / 76D / 103A / 104I / 159D / 213R / 232V / 236H / 245R / 260A; 68A / 76D / 103 A / 1041 / 159D / 210I / 232V / 236H / 245R / 26OA; 68A 103A 1041 / 159D / 1 3D / 232V7236H / 245R / 248D / 252K; 68A / 103 A 1041 / 159D / 209W / 232V / 236H / 245R; 68A / 76D / 103 A 1041 / 159D / 21 1 R / 232V / 236H / 245R; 68A / 76D / 103A 104I / 159D / 215R / 232V / 236H 245R; 68A / 103A / 104I / 159D / 213R / 232V / 236H / 245R / 260A; 68A / 76D / 103 A / 1041 / 159D / 232V / 236H / 245R; 68A / 103A 104I / 159D / 232V 236H / 245R / 252K; 68A / 103A / 104I / 159D / 232V / 236H 245R; 68A / 103A / 1041 / 159D / 232V / 236H / 245R / 257V; 68A / 103.A / 104I / 159D / 185D / 232V / 236H / 245R / 248D / 252K; 68A / 103 A / 1041 / 159D / 21 OL / 232 V / 236H / 245R / 248D / 252K; 68A / 103 A / 1041 / 159D / 185D 210I./232V/236H/245R/248D/252K; 68A / 103A / 104I / 159D / 213E? 232V / 236H / 245R / 248D / 2S2K; 68A / 103A / 104I / 159D / 230V / 232V / 236H / 245R; 68A / 76D / 103A / 104I / 159D / 209W / 232V / 236H / 245R; 68A / 103A / 104I / 232V / 236H / 245R / 248D / 257V / 275H; 68A / 103A / 104I / 232V / 236H 245R 2S7V / 275H; 68A / 103 A 1041 / 213E 232V / 236H / 245R / 248D / 252; 68A / 103 A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 68A / 103A / 104I / 159D / 2101 / 232V / 236H / 245R; 68A / 103A / 1041 / 159D / 210L 232V / 236H / 245R; 68A / 103A / 104I / 159D / 213G / 232V / 236H / 245R; 68A / 103 A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K / 270A; 76D / 103A / 1041 / 159D / 232V / 236H 245R; 76D / 103 to 1041/131 V / l 59D / 232V / 236H / 245R / 248D / 252K; 76D / 103A 104I / 159D / 213R 232V / 236H / 245R / 260A; 97E / 103A / 104I / 159D / 232V / 236H 245R 248D / 252K; 98L / 103A 104I / 159D / 232V / 236H / 245R / 248D / 252K; 98L / 102A / 1 O3A / 104I / 159D / 212G / 232V / 236H / 245R / 248D / 252K; 101G / 103A / 1041 / 159D / 232V / 236H / 245R / 248D / 252K; 102A / 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252K; 103A / 104I / 159D / 232V / 236H / 245R / 248D / 252; 103A 104I / 159D / 213R 232V / 236H / 245R / 248D / 252K: 10 103A / 104I / 130G / 159D / 232V / 236H / 245R / 248D / 252K; 103A / 104I / 159D / 217E 232V / 236H / 245R / 248D / 252K; 103 A / 1041 / 159D / 248D / 252K / 270V; 103 A / 1041 / 159D / 232V / 236H / 245R; 103A / 104I / 159D / 205I / 209W / 232V / 236H 245R; 103 A / 1041 / 159D / 232V / 236H / 245R / 257V; 103A / 1041 / 159D / 2051 / 209W / 232V / 236H / 245R / 257V; 103 A / 1041/131 V / l 59D / 232V / 236H / 245R / 248D / 252K; 103 A / 1041 / 159D / 205I / 209W / 210I / 232V / 236H / 245R 257V; and 15 103A / 1041 / 159D / 232V / 245R / 248D / 252K. SUMMARY OF THE INVENTION The present invention relates to cleaning compositions comprising a protease variant, a cleaning composition comprising a protease variant including a substitution of an amino acid residue with another natural amino acid residue at an amino acid residue position corresponding to position 103 of Bacillus amyloliquefaciens subtilisin in combination with a substitution of an amino acid residue with another natural amino acid residue at one or more amino acid residue positions corresponding to positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77.78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 111, 114, 116, 117, 119, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 21 1, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of Bacillus amyloliquefaciens subtilisin; wherein each said protease variant includes a substitution of amino acid residues at the positions corresponding to positions 103 and 76, there is also a substitution of an amino acid residue at one or more amino acid residue positions different from the residue positions of amino acids corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and one or more cleaning attachment materials; another cleaning composition comprises a protease variant, which includes a substitution of an amino acid residue with another amino acid residue that occurs naturally at one or more amino acid residue positions corresponding to positions 62, 212, 230, 232 , 252, and 257 of Bacillus amyloliquefaciens subtilisin; and one or more cleaning attachment materials; In addition, methods for using the cleaning compositions are provided. P00 / 586F RM / cgt *