HIGHLY ALKALINE TEXTILE DETERGENT WITH PROTEASE

Description

[0001] The invention lies in the field of detergents, in particular detergents containing at least one enzyme. The invention relates to a textile detergent, especially a liquid textile detergent, comprising at least one protease whose amino acid sequence has been modified, particularly with regard to its use in textile detergents, and at least one detergent ingredient, wherein the textile detergent has a pH value of approximately 9 to approximately 12. Also part of the invention are the corresponding washing processes, the use of the agents described herein, and the use of proteases in textile detergents with a pH value of approximately 9 to approximately 12, as well as the use of proteases in textile detergents with a pH value of approximately 9 to approximately 12 for the removal of protease-sensitive soils from textiles.

[0002] The use of enzymes in detergents has been established in the art for decades. They serve to expand the performance spectrum of the respective products according to their specific activities. These include, in particular, hydrolytic enzymes such as proteases, amylases, lipases, and cellulases. The first three hydrolyze proteins, starches, and fats, thus contributing directly to dirt removal. Cellulases are used especially for their tissue-cleaning properties. Another group of detergent enzymes are oxidative enzymes, especially oxidases, which, possibly in combination with other components, primarily serve to bleach soils or to neutralize the bleaching agents. in situIn addition to these enzymes, which are subject to continuous optimization, further enzymes are constantly being provided for use in detergents in order to optimally tackle special types of soiling, such as pectinases, β-glucanases, mannanases or other hemicellulases (glycosidases) for the hydrolysis of special plant polymers in particular.

[0003] The longest-established enzymes, and those found in virtually all modern, high-performance detergents, are proteases. They are therefore among the most technically important enzymes. Of these, subtilisin-type proteases (subtilases, subtilopeptidases, EC 3.4.21.62) are particularly important. These are serine proteases due to the catalytically active amino acids. They act as non-specific endopeptidases and hydrolyze any amide bonds located within peptides or proteins. Their optimum pH is usually in the alkaline range, around pH 9. An overview of this family is provided, for example, by the article "Subtilases: subtilisin-like Proteases" by R. Siezen, pages 75-95 in "Subtilisin enzymes," edited by R. Bott and C. Betzel, New York, 1996. subtilases are naturally produced by microorganisms. These include, in particular, those of Bacillus speciesThe most important group within the subtilisins is the formed and secreted subtilisins.

[0004] Examples of subtilisin-type proteases preferably used in detergents are the subtilisins BPN' and Carlsberg, the protease PB92, the subtilisins 147 and 309, the protease from Bacillus lentus, especially from Bacillus lentus DSM 5483, subtilisin DY and the enzymes thermitase, proteinase K and the proteases TW3 and TW7, which are no longer classified as subtilisins in the narrower sense, as well as variants of the aforementioned proteases that have an altered amino acid sequence compared to the original protease.

[0005] For example, European patent application EP 2016175 A1 describes a protease intended for use in washing and cleaning agents made of Bacillus pumilusThis has been revealed. Generally, only selected proteases are suitable for use in liquid surfactant-containing preparations. Many proteases do not exhibit sufficient catalytic performance in such preparations, or they are not sufficiently stable. Therefore, for the use of proteases in textile detergents, high catalytic activity and stability under conditions such as those encountered during a washing process are particularly desirable.

[0006] Solid detergents typically have a pH value of around 8 to 10, while liquid detergents can have a pH value of around 7 to 9. Alkaline liquid detergents are relatively widespread and particularly popular in North America. However, the alkaline pH makes it difficult to find suitable enzymes and incorporate them into alkaline liquid detergents, as the high pH can negatively affect not only the enzymes' performance but also their storage stability. Furthermore, enzymes are generally less stable in liquid detergents with a high water content.

[0007] Consequently, prior art liquid formulations containing protease and surfactants have the disadvantage that the proteases contained do not exhibit satisfactory proteolytic activity or are not sufficiently stable under standard washing conditions (e.g. in a temperature range of 20 to 40°C), especially at a higher pH value, and the formulations therefore do not show optimal cleaning performance on protease-sensitive soils.

[0008] One goal in the development of detergent formulations is therefore to modify the enzymes they contain, either selectively or randomly, using methods known from the prior art, and thus optimize them for use in detergents. These methods include, for example, point mutagenesis, deletion or insertion mutagenesis, or fusion with other proteins or protein fragments.

[0009] Although a number of proteases for detergents are known from the prior art, there is still a need for proteases for use in highly alkaline textile detergents, especially liquid textile detergents with a pH value of about 9 to about 12.

[0010] The task was therefore to further improve the stability of proteases in highly alkaline textile detergents and thus the cleaning performance of these detergents. Surprisingly, it has now been found that a protease, as defined herein, or a sufficiently similar protease (with respect to sequence identity), exhibits improved storage stability compared to conventional proteases and / or the wild-type form (SEQ ID NO:1) and is therefore particularly suitable for use in textile detergents with a pH of approximately 9 to 12.

[0011] The present invention therefore relates, in a first aspect, to a textile detergent comprising a) at least one protease, wherein the protease has proteolytic activity and comprises an amino acid sequence having at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its total length and, with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, amino acid substitutions, in particular the amino acid substitutions 9T, 130D, 130V, 133A, 144K, 217M, 252T and 271E, and (ii) at least one, preferably at least two, of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, at least one further amino acid substitution, in particular selected from 6W, 6F, 89A, 89G, 131H, 131Y, 131F, 166M, 166L, 166I, 189T, 189L, 189I, 211N, 211Q, 224A and 224G, preferably selected from 6W, 89A, 131H, 166M, 189T, 211N and 224A,comprising; and b) at least one detergent ingredient, preferably in an amount of 0.01 to 99.9 wt.%, wherein the textile detergent has a pH value of about 9 to about 12, measured in a 1 wt.% solution in deionized water at 20°C. Preferably, the detergent according to the invention is a liquid textile detergent. More preferably, the detergent according to the invention has a pH value in the range of 9 to 12, in particular 9.5 to 11.5, more preferably 10 to 11, and most preferably pH 10, in a 1 wt.% solution in deionized water at 20°C.

[0012] WO2019 / 048495 teaches protease variants from B. pumilus,in particular a variant with the mutations 9T, 130D, 133A, 144K, 217M, 252T and 271E (D5, mutant 38). The subject matter of the present application differs from the teaching of WO2019 / 048495 by the mutations mentioned under (ii) and the pH value (present application: pH 9-12; WO2019 / 048495: no pH is mentioned in the example). The additional mutations at positions 6, 89, 131, 166, 189, 211 or 224 (preferably selected from 6W / F, 89A / G, 131H / Y / F, 166M / L / I, 189T / L / I, 211N / Q and 224A / G) result in improved storage stability at high pH (see page 3, paragraphs 1-4 of the originally submitted description; example).

[0013] Textile detergents according to the invention exhibit performance advantages over other textile detergents, particularly when they contain at least one additional enzyme of the same or a different type, such as amylase, cellulase, lipase, mannanase, or pectinase (this list of further enzymes is not exhaustive). Therefore, it is preferred that the textile detergents according to the invention contain at least one additional enzyme of the same type (i.e., another protease) or a different type. Preferably, the detergent comprises at least one amylase, wherein the amylase has amylolytic activity and is selected from a) an α-amylase comprising an amino acid sequence that is at least 80% identical over its total length to the amino acid sequence specified in SEQ ID NO:2 and optionally has at least one amino acid substitution at one of positions 172, 202, 208, 255, and 261 in the numbering according to SEQ ID NO:2.preferably selected from the group consisting of M202L, M202V, M202S, M202T, M202I, M202Q, M202W, S255N, R172Q and combinations thereof; and / or b) an α-amylase comprising an amino acid sequence that is at least 60% identical over its total length to the amino acid sequence specified in SEQ ID NO:3 and optionally includes at least one amino acid substitution at one of positions 9, 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 195, 202, 203, 214, 231, 256, 257, 258, 269, 270, 272, 283, 295, 296, 298, 299, 303, 304, 305, 311, 314, 315, 318, 319, 320, 323, 339, 345, 361, 378, 383, 419, 421, 437, 441, 444, 445, 446, 447, 450, 458, 461, 471, 482 and 484 and / or a deletion at one of positions 183 and 184 in the numbering according to SEQ ID NO:3, preferably at least one amino acid substitution at one of positions 9, 26, 149, 182, 186, 202, 257, 295, 299, 323, 339 and 345,and / or particularly preferably at least one amino acid substitution or deletion selected from the group consisting of R118K, D183*, G184*, N195F, R320K, R458K and combinations thereof; and / or c) an α-amylase comprising an amino acid sequence that is at least 90% identical over its total length to the amino acid sequence specified in SEQ ID NO:4 and optionally includes at least one substitution and / or deletion at one of positions 93, 116, 118, 129, 133, 134, 140, 142, 146, 147, 149, 151, 152, 169, 174, 183, 184, 186, 189, 193, 195, 197, 198, 200, 203, 206, 210, 212, 213, 235, 243, 244, 260, 262, 284, 303, 304, 320, 338, 347, 359, 418, 431, 434, 439, 447, 458, 469, 476 and 477 in the numbering according to SEQ ID NO:4, exhibiting, preferably amino acid deletions at positions 183 and 184. Such amylases are known from, for example, WO2013 / 063460 and EP2357220.

[0014] Another object of the invention is a method for producing such a textile detergent and a method for cleaning textiles in which a detergent according to the invention is used.

[0015] Another aspect of the invention is the use of such a textile detergent for cleaning textiles.

[0016] A further object of the invention is the use of a protease comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its total length and, with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, amino acid substitutions, in particular the amino acid substitutions 9T, 130D, 130V, 133A, 144K, 217M, 252T and 271, and (ii) at at least one, preferably at least two, of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, at least one further amino acid substitution, in particular selected from 6W, 6F, 89A, 89G, 131H, 131Y, 131F, 166M, 166L, 166I, 189T, 189L, 1891, 211N, 211Q, 224A and 224G, preferably selected from 6W, 89A, 131H, 166M, 189T, 211N and 224A, comprising, in a textile detergent, preferably a liquid textile detergent,for the removal of protease-sensitive soiling from textiles, wherein the textile detergent has a pH value of approximately 9 to approximately 12, measured in a 1 wt% solution in deionized water at 20°C.

[0017] Particularly preferred in the means, methods and uses according to the invention is a protease which has at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its total length and, with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, contains the amino acid substitutions P9T, N130D, T133A, N144K, Y217M, N252T and Q271E, and (ii) at at least two of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, contains at least two further amino acid substitutions consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, G166M, G166L, G166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G, preferably Y6W, S89A, G131H, G166M, S189T, S211N and S224A, existing group selected, has.

[0018] These and other aspects, features, and advantages of the invention will become apparent to the person skilled in the art upon studying the following detailed description and claims. Each feature from one aspect of the invention can be incorporated into any other aspect of the invention. Furthermore, it is understood that the examples contained herein are intended to describe and illustrate the invention, but do not limit it, and in particular, the invention is not limited to these examples.

[0019] Unless otherwise stated, all percentages are weight percent (wt%).

[0020] Numeric ranges specified in the format "from x to y" include the stated values. If multiple preferred numeric ranges are specified in this format, it is understood that all ranges resulting from the combination of the different endpoints are also included.

[0021] "At least one", as used herein, means one or more, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more.

[0022] The term "detergent", as used herein, is synonymous with the term "textile detergent" or "agent" and refers to a composition for cleaning textiles as explained in the description.

[0023] "Approximately", "about" or "about", as used herein in reference to a numerical value, refer to the corresponding numerical value ±10%, preferably ±5%.

[0024] "Liquid," as used herein, includes liquids and gels as well as pasty compositions. It is preferred that the liquid compositions are free-flowing and pourable at room temperature, but it is also possible that they exhibit a yield point.

[0025] "Highly alkaline", as used herein, means a pH value of about 9 to about 12, measured in 1 wt% solution in deionized water at 20°C.

[0026] The present invention is based on the surprising finding of the inventors that amino acid substitutions at the positions described herein result in improved storage stability of this modified protease in highly alkaline detergents compared to conventional proteases and / or compared to the wild type.

[0027] This is particularly surprising insofar as no such proteases or amino acid substitutions have previously been associated with improved storage stability of the corresponding protease in highly alkaline detergents, especially liquid highly alkaline detergents.

[0028] In preferred embodiments, the modification(s) of the proteases used according to the invention, in each case with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, and (ii) at at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, leads to improved storage stability of this modified protease in highly alkaline textile detergents (pH 9 to 12).

[0029] In further preferred embodiments of the proteases used according to the invention, the protease, in each case with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, has the amino acid substitutions P9T, N130D, T133A, N144K, Y217M, N252T and Q271E and (ii) at at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, has at least one amino acid substitution consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G existing group is selected.

[0030] In preferred embodiments, the protease used according to the invention, with reference to the numbering according to SEQ ID NO:1, (i) at positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, has the amino acid substitutions P9T, N130D, T133A, N144K, Y217M, N252T and Q271E and (ii) at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, has at least one amino acid substitution consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, selected from the group consisting of S211N, S211Q, S224A and S224G, the combination of the amino acid substitutions from group (i) and the at least one amino acid substitution from group (ii) leads to improved storage stability of this modified protease in highly alkaline textile detergents (pH 9 to 12).

[0031] In particularly preferred embodiments, the detergent according to the invention contains a protease with one of the following amino acid substitution variants: (i) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+N252T+Q271E; (ii) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+S224A+N252T+Q271E; (iii) P9T+S89A+N130D+G131H+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (iv) Y6W+P9T+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (v) P9T+N130D+T133A+N144K+G166M+S211N+Y217M+N252T+Q271E; (vi) P9T+S89A+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E, where the numbering refers to the numbering according to SEQ ID NO:1.

[0032] Certain embodiments of the proteases used according to the invention have improved storage stability. They exhibit increased stability in highly alkaline textile detergents compared to conventional proteases and / or the wild-type enzyme (SEQ ID NO:1), particularly during storage of 3 or more days, 4 or more days, 7 or more days, 10 or more days, 12 or more days, 14 or more days, 21 or more days, or 28 or more days, wherein the textile detergent has a pH of approximately 9 to approximately 12, measured in a 1 wt% solution in deionized water at 20°C.

[0033] Certain embodiments of the proteases according to the invention can exhibit increased catalytic activity in highly alkaline textile detergents, independently of or in addition to their increased storage stability. In various embodiments, the proteases used according to the invention can possess a proteolytic activity of at least 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, or 110%, relative to the wild type (SEQ ID NO:1). Such performance-enhanced proteases enable improved washing results on protease-sensitive soils in various temperature ranges, particularly a temperature range of 20 to 40°C.

[0034] Furthermore, preferred embodiments of the proteases used according to the invention exhibit particular stability in detergents, for example, against surfactants and / or bleaching agents and / or chelators, and / or against temperature influences, especially against high temperatures, for example between 50 and 65°C, particularly 60°C, and / or against pH changes and / or against denaturing or oxidizing agents and / or against proteolytic degradation and / or against changes in redox conditions. Thus, particularly preferred embodiments of the invention provide performance-enhanced and / or temperature-stable protease variants. Further particularly preferred embodiments of the invention provide performance-enhanced and temperature-stable protease variants.Such advantageous embodiments of the proteases according to the invention therefore enable improved washing results on protease-sensitive soils in a wide temperature range.

[0035] The proteases used according to the invention exhibit enzymatic activity, i.e., they are capable of hydrolyzing peptides and proteins, particularly in detergents. A protease used according to the invention is therefore an enzyme that catalyzes the hydrolysis of amide / peptide bonds in protein / peptide substrates and is thus able to cleave proteins or peptides. Furthermore, a protease used according to the invention is preferably a mature protease, i.e., the catalytically active molecule without signal and / or propeptide(s). Unless otherwise specified, the given sequences also refer to mature (processed) enzymes.

[0036] In various embodiments of the invention, the protease used according to the invention is a free enzyme. This means that the protease can interact directly with all components of a composition and, if the composition is a liquid, that the protease is in direct contact with the solvent of the composition (e.g., water). In other embodiments, a composition may contain proteases that form an interaction complex with other molecules or that contain a "coating." In this case, a single or multiple protease molecules may be separated from the other components of the composition by a surrounding structure. Such a separating structure can be formed by, but is not limited to, vesicles, such as a micelle or a liposome. The surrounding structure may also be a virus particle, a bacterial cell, or a eukaryotic cell. In various embodiments, a composition may contain cells of Bacillus pumilus or Bacillus gibsonii or Bacillus subtilis, which express the proteases according to the invention, or contain cell culture supernatants of such cells.

[0037] In preferred embodiments of the invention, the protease used according to the invention comprises an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:1 over its total length to at least 70% and increasingly preferably to at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5% and 97%, in each case with respect to the numbering according to SEQ ID NO:1 (i) has the amino acid substitutions P9T, N130D, T133A, N144K, Y217M, N252T and Q271E, and (ii) has at least one amino acid substitution at at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, wherein the at least one amino acid substitution is selected from the group consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G.

[0038] In preferred embodiments of the invention, the protease used according to the invention comprises an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:1 over its total length to at least 70% and increasingly preferably to at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96% and 96.5%, and in each case, with reference to the numbering according to SEQ ID NO:1 (i) the (ii) has amino acid substitutions P9T, N130D, T133A, N144K, Y217M, N252T and Q271E, and (ii) has at least two amino acid substitutions at at least two of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, wherein the at least two amino acid substitutions are selected from the group consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G.

[0039] In preferred embodiments of the invention, the protease used according to the invention comprises an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:1 over its total length to at least 70% and increasingly preferably to at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5% and 96%, and in each case, with reference to the numbering according to SEQ ID NO:1 (i), the amino acid substitutions (i) comprises P9T, N130D, T133A, N144K, Y217M, N252T and Q271E, and (ii) comprises at least three amino acid substitutions at at least three of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, wherein the at least three amino acid substitutions are selected from the group consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G.

[0040] In preferred embodiments of the invention, the protease used according to the invention comprises an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:1 over its total length to at least 70% and increasingly preferably to at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5% and 96%, and in each case, with reference to the numbering according to SEQ ID NO:1 (i), the amino acid substitutions (i) comprises P9T, N130D, T133A, N144K, Y217M, N252T and Q271E, and (ii) comprises at least four amino acid substitutions at at least four of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, wherein the at least four amino acid substitutions are selected from the group consisting of Y6W, Y6F, S89A, S89G, G131H, G131Y, G131F, S166M, S166L, S166I, S189T, S189L, S189I, S211N, S211Q, S224A and S224G.

[0041] In particularly preferred embodiments, the protease used according to the invention contains one of the following amino acid substitution variants: (i) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+N252T+Q271E; (ii) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+S224A+N252T+Q271E; (iii) P9T+S89A+N130D+G131H+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (iv) Y6W+P9T+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (v) P9T+N130D+T133A+N144K+G166M+S211N+Y217M+N252T+Q271E; (vi) P9T+S89A+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E, where the numbering refers to the numbering according to SEQ ID NO:1.

[0042] In the context of the present invention, the feature that a protease has at least one of the specified amino acid substitutions means that it contains one (of the specified) amino acid substitution(s) at the respective position, i.e., at least the specified positions are not otherwise mutated or deleted, for example by fragmentation of the protease.

[0043] The identity of nucleic acid or amino acid sequences is determined by sequence comparison. This sequence comparison is based on the BLAST algorithm, which is established in the art and commonly used (see, e.g., Altschul et al. (1990) "Basic local alignment search tool", J. Mol. Biol. 215:403-410 and Altschul et al. (1997) "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res., 25:3389-3402). In principle, it is achieved by matching similar sequences of nucleotides or amino acids in the nucleic acid or amino acid sequences. A tabular assignment of the relevant positions is called an alignment. Another algorithm available in the art is the FASTA algorithm. Sequence comparisons (alignments), especially multiple sequence comparisons, are performed using computer programs. Frequently used programs include, for example, the Clustal series (see, e.g., Chenna et al.).(2003) "Multiple sequence alignment with the Clustal series of programs", Nucleic Acid Res. 31:3497-3500), T-Coffee (see, e.g., Notredame et al. (2000) "T-Coffee: A novel method for multiple sequence alignments", J. Mol. Biol. 302:205-217) or programs based on these programs or algorithms. Sequence comparisons (alignments) are also possible using the computer program Vector NTI® Suite 10.3 (Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California, USA) with the predefined standard parameters, whose AlignX module for sequence comparisons is based on ClustalW. Unless otherwise specified, the sequence identity given herein is determined using the BLAST algorithm.

[0044] Such a comparison also allows for a statement about the similarity of the compared sequences to one another. This is usually expressed as percent identity, i.e., the proportion of identical nucleotides or amino acid residues at the same positions or positions corresponding to each other in an alignment. The broader concept of homology includes conserved amino acid substitutions in the analysis of amino acid sequences, i.e., amino acids with similar chemical activity, since these usually exert similar chemical activities within the protein. Therefore, the similarity of the compared sequences can also be expressed as percent homology or percent similarity. Identity and / or homology statements can be made for entire polypeptides or genes, or only for individual regions. Homologous or identical regions of different nucleic acid or amino acid sequences are therefore defined by similarities in the sequences.Such regions often exhibit identical functions. They can be small, comprising only a few nucleotides or amino acids. Often, these small regions perform essential functions for the overall activity of the protein. It can therefore be advantageous to refer to sequence similarities only in individual, possibly small, regions. Unless otherwise stated, however, statements of identity or homology in this application refer to the total length of the respective nucleic acid or amino acid sequence.

[0045] In the context of the present invention, the statement that an amino acid position corresponds to a numerically designated position in SEQ ID NO:1 therefore means that the corresponding position is assigned to the numerically designated position in SEQ ID NO:1 in an alignment as defined above.

[0046] For the description of substitutions affecting exactly one amino acid position (amino acid exchanges), the following convention is used: first, the naturally occurring amino acid is designated using the internationally accepted one-letter code, followed by the corresponding sequence position, and finally the inserted amino acid. Multiple exchanges within the same polypeptide chain are separated by slashes. In the case of insertions, additional amino acids are named after the sequence position. In the case of deletions, the missing amino acid is replaced by a symbol, such as an asterisk or a dash, or a Δ is indicated before the corresponding position. For example, A95G describes the substitution of alanine at position 95 by glycine, A95AG the insertion of glycine after the amino acid alanine at position 95, and A95* or ΔA59 the deletion of alanine at position 95.This nomenclature is familiar to experts in the field of enzyme technology.

[0047] Another object of the present invention is a detergent comprising a protease, characterized in that the protease is obtainable from a protease according to the invention as a starting molecule by one or more conservative amino acid substitutions. The term "conservative amino acid substitution" means the exchange (substitution) of one amino acid residue for another amino acid residue, wherein this exchange does not lead to a change in polarity or charge at the position of the exchanged amino acid, e.g., the exchange of one nonpolar amino acid residue for another nonpolar amino acid residue. Conservative amino acid substitutions within the scope of the invention include, for example: G=A=S, I=V=L=M, D=E, N=Q, K=R, Y=F, S=T, G=A=I=V=L=M=Y=F=W=P=S=T.

[0048] Another object of the present invention is a detergent comprising a protease, characterized in that the protease is obtainable from a protease according to the invention as a starting molecule by fragmentation, deletion, insertion or substitution mutagenesis and comprises an amino acid sequence that is identical to the starting molecule over a length of at least 190, 200, 210, 220, 230, 240, 250, 260, 270, 271, 272, 273, 274 or 275 contiguous amino acids, wherein the protease (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 or 271, and (ii) at least one amino acid substitution at at least one of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224.

[0049] For example, it is possible to delete individual amino acids at the termini or in the loops of the enzyme without losing or reducing its proteolytic activity. Furthermore, such fragmentation, deletion, insertion, or substitution mutagenesis can, for example, reduce the allergenicity of the enzymes in question and thus improve their overall usability. Advantageously, the enzymes retain their proteolytic activity even after mutagenesis; that is, their proteolytic activity is at least equal to that of the original enzyme. In a preferred embodiment, the proteolytic activity is at least 80%, preferably at least 90%, of the activity of the original enzyme. Further substitutions can also have beneficial effects. Both single and multiple contiguous amino acids can be exchanged for other amino acids.

[0050] The amino acid positions are determined by aligning the amino acid sequence of a protease according to the invention with the amino acid sequence of the protease from Bacillus pumilus, as defined in SEQ ID NO:1. Furthermore, the assignment of positions is based on the mature protein. This assignment is particularly applicable if the amino acid sequence of a protease according to the invention comprises a higher or lower number of amino acid residues than the protease from Bacillus pumilus according to SEQ ID NO:1. Based on the mentioned positions in the amino acid sequence of the protease from Bacillus pumilus The change positions in a protease according to the invention are those that are assigned to these positions in an alignment.

[0051] Further confirmation of the correct assignment of the amino acids to be modified, i.e., in particular their functional correspondence, can be provided by comparative experiments in which the two positions assigned to each other on the basis of an alignment are modified in the same way in both compared proteases, and it is observed whether the enzymatic activity is altered in the same way in both. If, for example, an amino acid substitution at a specific position of the protease according to SEQ ID NO:1 is accompanied by a change in an enzymatic parameter, for example, an increase in the KM value, and a corresponding change in the enzymatic parameter, e.g., also an increase in the KM value, is observed in a protease variant according to the invention, whose amino acid substitution was achieved by the same introduced amino acid, then this can be seen as confirmation of the correct assignment.

[0052] A protease according to the invention can be additionally stabilized, in particular by one or more mutations, e.g., substitutions, or by coupling to a polymer. Increased stability during storage and / or use, e.g., during the washing process, results in longer-lasting enzymatic activity and thus improved cleaning performance. In principle, all stabilization methods described in the prior art and / or deemed suitable are suitable. Stabilization achieved through mutations of the enzyme itself is preferred, as such stabilization does not require any further processing steps after enzyme production. Other stabilization methods include, for example: Alteration of the binding of metal ions, especially the calcium binding sites, e.g. by exchanging one or more of the amino acid(s) involved in the calcium binding for one or more negatively charged amino acids and / or by introducing sequence changes in at least one of the sequences of the two amino acids arginine / glycine; protection against the influence of denaturing agents such as surfactants by mutations that cause a change in the amino acid sequence on or at the surface of the protein; exchange of amino acids located near the N-terminus for those that presumably interact with the rest of the molecule via non-covalent interactions and thus contribute to maintaining the globular structure.

[0053] Preferred embodiments are those in which the enzyme is stabilized in several ways, since multiple stabilizing mutations act additively or synergistically.

[0054] Another object of the invention is a protease as described above, characterized in that it has at least one chemical modification. A protease with such a modification is called a derivative, i.e., the protease is derivatized. For the purposes of this application, derivatives are therefore understood to be proteins whose pure amino acid chain has been chemically modified. Such derivatizations can eg in vivo Derivatization occurs through the host cell that expresses the protein. In this regard, couplings of low-molecular-weight compounds such as lipids or oligosaccharides are particularly noteworthy. However, derivatizations can also occur. in vitroDerivatization can be carried out, for example, by the chemical modification of a side chain of an amino acid or by covalently binding another compound to the protein. For instance, coupling amines to carboxyl groups of an enzyme is possible to alter the isoelectric point. Such another compound can also be a further protein, which is bound to a protein according to the invention, for example, via bifunctional chemical bonds. Similarly, derivatization refers to covalent binding to a macromolecular support or non-covalent inclusion in suitable macromolecular cage structures. Derivatizations can, for example, influence the substrate specificity or the binding strength to the substrate, or cause a temporary blockage of enzymatic activity if the attached substance is an inhibitor. This can be useful, for example, during storage.Such modifications can also influence the stability or enzymatic activity. They can also serve to reduce the allergenicity and / or immunogenicity of the protein and thus, for example, increase its skin compatibility. For instance, coupling with macromolecular compounds, e.g., polyethylene glycol, can improve the protein with regard to stability and / or skin compatibility. In the broadest sense, preparations of a protein according to the invention can also be understood as derivatives of that protein. Depending on the method of extraction, processing, or preparation, a protein may be associated with various other substances, e.g., from the culture of the producing microorganisms. A protein may also have been deliberately combined with other substances, e.g., to increase its storage stability. Therefore, all preparations of a protein according to the invention are also considered to be in accordance with the invention.This is also independent of whether it actually exhibits this enzymatic activity in a particular preparation or not. It may be desirable for it to have no or only slight activity during storage and only develop its enzymatic function at the time of use. This can be controlled, for example, by using appropriate accompanying substances. In particular, the combined preparation of proteases with specific inhibitors is possible in this regard.

[0055] Of all the proteases or protease variants and / or derivatives described above, those whose storage stability and / or purification performance is improved compared to the starting variant are particularly preferred within the scope of the present invention, the purification performance being determined in a washing system as described below. Numerous proteases, and especially subtilisins, are formed as so-called preproteins, i.e., together with a propeptide and a signal peptide. The function of the signal peptide is typically to ensure the release of the protease from the cell producing it into the periplasm or the medium surrounding the cell, and the propeptide is usually necessary for the correct folding of the protease. The signal peptide and the propeptide are generally the N-terminal part of the preprotein.Under natural conditions, the signal peptide is cleaved from the rest of the protease by a signal peptidase. Subsequently, the protease, aided by the propeptide, correctly folds to its final position. The protease is then in its active form and cleaves the propeptide itself. After the propeptide is cleaved, the mature protease, particularly subtilisin, exerts its catalytic activity without the originally present N-terminal amino acids. For technical applications in general, and particularly within the scope of the invention, mature proteases, i.e., the processed enzymes after their synthesis, are preferred over preproteins. Furthermore, the proteases can be modified by the cells that produce them after the polypeptide chain has been synthesized, e.g., by the addition of sugar molecules, formylation, amination, etc.Such modifications are post-translational modifications and may, but do not necessarily, have an influence on the function of the protease.

[0056] "Variant," as used herein, refers to naturally occurring or artificially generated variations of a native protease that exhibits an amino acid sequence modified from the reference form. In addition to the amino acid modifications described above, proteases according to the invention may exhibit further amino acid modifications, in particular amino acid substitutions, insertions, or deletions. Such proteases are further developed, for example, by targeted genetic modification, i.e., by mutagenesis techniques, and optimized for specific applications or with regard to special properties (e.g., their catalytic activity, stability, etc.). Furthermore, nucleic acids according to the invention can be introduced into recombination reactions and thus used to generate entirely novel proteases or other polypeptides. The aim is to introduce targeted mutations, such as substitutions, insertions, or deletions, into known molecules in order to, for example,to improve the cleaning performance of enzymes. This can be achieved, in particular, by modifying the surface charges and / or the isoelectric point of the molecules and thereby their interactions with the substrate. For example, the net charge of the enzymes can be altered to influence substrate binding, especially for use in detergents and cleaning agents. Alternatively or additionally, one or more appropriate mutations can increase the stability or catalytic activity of the enzyme and thereby improve its cleaning performance. Advantageous properties of individual mutations, e.g., individual substitutions, can be complementary. A protease already optimized with respect to certain properties can therefore be further developed within the scope of the invention, e.g., with respect to its stability against surfactants and / or bleaching agents and / or other components.

[0057] A detergent according to the invention increasingly preferably contains the protease in an amount of 1 x 10 -8 to 5 wt.%, from 0.0001 to 1 wt.%, from 0.0005 to 0.5 wt.%, from 0.001 to 0.1 wt.%, in each case based on active protein and based on the total weight of the detergent.

[0058] In a further embodiment of the invention, the protease is characterized in that its cleaning performance (after storage, e.g. over 2 weeks) is not significantly reduced compared to conventional proteases and / or the wild type enzyme (SEQ ID NO:1), i.e. it has at least 80% of the reference washing performance, preferably at least 100%, more preferably at least 110% or more.

[0059] Within the scope of the invention, cleaning performance is understood to mean the ability of an agent to partially or completely remove existing soiling, in particular its brightening performance on one or more soilings on textiles. Examples of such soiling are blood on cotton, chocolate-milk / soot on cotton, cocoa on cotton, or porridge on cotton. Within the scope of the invention, both the detergent comprising the protease, or the washing solution formed by this agent, and the protease itself exhibit a cleaning performance. The cleaning performance of the protease thus contributes to the cleaning performance of the agent or the washing solution formed by the agent. The cleaning performance is preferably determined as described below.

[0060] The term "wash liquor" refers to the solution containing the detergent that comes into contact with the textiles or fabrics, thus interacting with the soiling present on them. The wash liquor is typically created when the washing process begins and the detergent is diluted with water, for example, in a washing machine or other suitable container.

[0061] The cleaning performance can be determined in a washing system containing a detergent at a dosage of 2.0 to 8.0 grams per liter of wash liquor, as well as the protease. The proteases to be compared are used at the same concentration (relative to the active protein). Using the same activity level for each protease ensures that even if the ratio of active substance to total protein (the specific activity values) differs, the respective enzymatic properties, such as the cleaning performance on specific types of soil, can be compared. Generally, a low specific activity can be compensated for by adding a larger amount of protein. Furthermore, the enzymes to be tested can also be used in the same amount of substance or weight if they exhibit different affinities for the test substrate in an activity test.In this context, the term "equal amount of substance" refers to using the enzymes under investigation in the same number of moles. The term "equal amount by weight" refers to using the enzymes under investigation in the same weight.

[0062] The concentration of the protease in the detergent intended for this washing system is 0.0001 to 0.1 wt.%, in particular 0.001 to 0.1 wt.%, more preferably 0.01 to 0.06 wt.% and particularly preferably 0.001 to 0.02 wt.%, based on active protein.

[0063] A preferred liquid detergent for such a washing system is composed as follows (all values ​​in weight percent): <1% antifoaming agent, 1-4% citric acid, 0.5-3% glycerin, 0.3-2% NaOH, 4-8% 1,2-propanediol, 2-5% FAEOS (fatty alcohol ether sulfate), 5-9% non-ionic surfactants (FAEO), 7-12% anionic surfactants (LAS), 0.5-1.5% protease stabilizer, 1-3% palm kernel oil fatty acids, 0.5-2% HEDP (1-hydroxyethane-(1,1-diphosphonic acid)), 2-6% monoethanolamine (MEA), 0.2-1% soil release polymer, remainder demineralized water. Preferably, the dosage of the liquid detergent is between 2.0 and 8.0 grams per liter of wash liquor, e.g., 2.5 g / L, 3.2 g / L, 3.5 g / L, 4.0 g / L, 4.7 g / L, 4.9 g / L, 5.5 g / L, or 5.9 g / L of wash liquor, or approximately 55 g / job. Washing is preferably carried out at a pH of approximately 9 to approximately 12, more preferably at approximately 10 to 11, and particularly preferably at pH 10.

[0064] The degree of whiteness, i.e., the lightening of the soiling, as a measure of cleaning performance, is preferably determined using optical measurement methods, preferably photometrically. A suitable device for this purpose is, for example, the Minolta CM508d spectrometer. Typically, the devices used for the measurement are calibrated beforehand with a white standard, preferably a supplied white standard.

[0065] Methods for determining protease activity are familiar to those skilled in the field of enzyme technology and are routinely used by them. For example, such methods are disclosed in Surfactants, Volume 7 (1970), pp. 125-132. Alternatively, protease activity can be determined via the release of the chromophore para-nitroaniline (pNA) from the substrate suc-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (AAPF). The protease cleaves the substrate and releases pNA. The release of pNA causes an increase in absorbance at 410 nm, the time course of which is a measure of the enzymatic activity (see Del Mar et al., 1979). The measurement is performed at a temperature of 25°C, pH 8.6, and a wavelength of 410 nm. The measurement time is 5 minutes and the measurement interval is 20 to 60 seconds. Protease activity is usually expressed in protease units (PE). Suitable protease activities are, for example, 2.25, 5, or 10 PE per ml of washing solution.However, protease activity is not zero.

[0066] An alternative test for determining the proteolytic activity of the proteases according to the invention is an optical measurement method, preferably a photometric method. The suitable test comprises the protease-dependent cleavage of the substrate protein casein. This is cleaved by the protease into a multitude of smaller subproducts. The totality of these subproducts exhibits increased absorption at 290 nm compared to uncleaved casein, whereby this increased absorption can be determined using a photometer, and thus a conclusion can be drawn about the enzymatic activity of the protease.

[0067] The protein concentration can be determined using known methods, e.g., the BCA method (bicinchoninic acid; 2,2'-bicinolyl-4,4'-dicarboxylic acid) or the biuret method (Gornall et al. (1948) J. Biol. Chem., 177:751-766). The determination of the active protein concentration can be carried out by titration of the active sites using a suitable irreversible inhibitor and determination of the residual activity (Bender et al. (1966) J. Am. Chem. Soc. 88(24):5890-5913).

[0068] Preferred embodiments of the proteases according to the invention achieve such advantageous cleaning performance even at low temperatures, particularly in the temperature ranges between 10 and 60°C, preferably between 15 and 50°C and especially preferably between 20 and 40°C.

[0069] According to the invention, the term "detergent" encompasses all conceivable types of detergents, both concentrates and undiluted products, for use on a commercial scale, in washing machines, or for hand washing or cleaning. This includes, for example, detergents for textiles, carpets, or natural fibers, for which the term "detergent" is used. Detergents within the scope of the invention also include washing aids that are added to the actual detergent during manual or machine textile washing to achieve an additional effect. Furthermore, textile pre- and post-treatment agents, i.e., agents with which the garment is brought into contact before the actual washing, are also considered detergents within the scope of the invention.These include products for loosening stubborn stains, and also those that, in a subsequent step of the actual textile washing process, impart further desirable properties to the laundry, such as a pleasant feel, wrinkle resistance, or low static charge. Fabric softeners are among the latter.

[0070] The detergents according to the invention, which can be in the form of powdered or granular solids, in compacted or post-compacted particle form, as homogeneous solutions or suspensions, can contain, in addition to a protease according to the invention, all known ingredients customary in such agents, wherein the detergent preferably contains such an additional ingredient in an amount of 0.01 to 99.9% by weight. The detergents according to the invention can, in particular, contain surfactants, builders, polymers, glass corrosion inhibitors, corrosion inhibitors, bleaching agents such as peroxygen compounds, bleach activators, or bleach catalysts.Furthermore, they may contain water-miscible organic solvents, other enzymes, enzyme stabilizers, sequestrants, electrolytes, pH regulators and / or other auxiliary substances such as optical brighteners, graying inhibitors, color transfer inhibitors, foam regulators, as well as colorants and perfumes and combinations thereof.

[0071] Suitable surfactants include, in particular, anionic surfactants, nonionic surfactants, and mixtures thereof. The compositions according to the invention can contain one or more surfactants, including, in particular, anionic surfactants, nonionic surfactants, and mixtures thereof, but also cationic, zwitterionic, and / or amphoteric surfactants. The compositions preferably contain 5 to 70 wt.% surfactant, more preferably 5 to 55 wt.%, and more preferably 5 to 35 wt.% surfactant. Particularly preferably, the washing compositions according to the invention contain no more than 30 wt.% surfactants, and most preferably no more than 20 wt.% surfactants.

[0072] Suitable anionic surfactants are, in particular, soaps and those containing sulfate or sulfonate groups with preferably alkali ions as cations. Usable soaps are preferably the alkali salts of saturated or unsaturated C12-18 fatty acids. Such fatty acids can also be used in a partially neutralized form. Suitable sulfate-type surfactants include the salts of the sulfuric acid half-esters of C12-18 fatty alcohols and the sulfation products of the aforementioned nonionic surfactants with a low degree of ethoxylation. Usable sulfonate-type surfactants include, for example, C9-14 alkylbenzenesulfonates, alkanesulfonates obtained from C12-18 alkanes by sulfochlorination or sulfoxidation followed by hydrolysis or neutralization, C12-18 olefin sulfonates formed by the reaction of corresponding monoolefins with sulfur trioxide, mixtures of alkene and hydroxyalkanesulfonates, and disulfonates, such as those found in...obtained from C 12-18 monoolefins with terminal or internal double bonds by sulfonation with gaseous sulfur trioxide and subsequent alkaline or acidic hydrolysis of the sulfonation products, as well as α-sulfofaticial esters (ester sulfonates) that are formed by the sulfonation of fatty acid methyl or ethyl esters, e.g. α-sulfonated methyl esters of hydrogenated coconut, palm kernel or tallow fatty acids.

[0073] Preferably, the composition comprises 2 to 55 wt.%, more preferably 3 to 35 wt.%, of anionic surfactant. Most preferably, the composition comprises 3 to 15 wt.% of alkylbenzenesulfonate. Furthermore, the composition may preferably contain other anionic surfactants, in particular alkyl ether sulfates, as well as non-ionic surfactants, in particular fatty alcohol alkoxylates. These may then constitute the remainder of the surfactants.

[0074] Suitable alkylbenzenesulfonates are preferably selected from linear or branched alkylbenzenesulfonates of the formula In which R' and R" are independently H or alkyl and together contain 6 to 19, preferably 7 to 15 and particularly 9 to 13 carbon atoms. A particularly preferred representative is sodium dodecylbenzylsulfonate.

[0075] The alkali and, in particular, the sodium salts of the sulfuric acid half-esters of C 12-18 fatty alcohols, e.g., from coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol, or of C 10-20 oxo alcohols, and those half-esters of secondary alcohols of these chain lengths are preferred as alk(en)yl sulfates. Also preferred are alk(en)yl sulfates of the aforementioned chain lengths which contain a synthetic, petrochemically produced, straight-chain alkyl group and which exhibit analogous degradation behavior to the corresponding compounds based on fatty chemical raw materials. For detergent applications, C 12-16 alkyl sulfates, C 12-15 alkyl sulfates, and C 14-15 alkyl sulfates are preferred.

[0076] Sulfuric acid monoesters of straight-chain or branched C 7-21 alcohols ethoxylated with 1 to 6 mol of ethylene oxide, such as 2-methyl-branched C 9-11 alcohols with an average of 3.5 mol of ethylene oxide (EO) or C 12-18 fatty alcohols with 1 to 4 EO, are also suitable.

[0077] Suitable alkyl ether sulfates are, for example, compounds of the formula R< -O-(AO) n -SO 3 -< X +< . In this formula, R< represents a linear or branched, substituted or unsubstituted alkyl group, preferably a linear, unsubstituted alkyl group, and particularly preferably a fatty alcohol group. Preferred R< groups are selected from decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, and eicosyl groups and mixtures thereof, with those having an even number of carbon atoms being preferred. Particularly preferred residues R1< are derived from C12-18 fatty alcohols, e.g., coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl, or stearyl alcohol, or from C10-20 oxo alcohols. AO stands for an ethylene oxide (EO) or propylene oxide (PO) group, preferably an ethylene oxide group. The index n stands for an integer from 1 to 50, preferably from 1 to 20, and particularly from 2 to 10.Most preferably, n represents the numbers 2, 3, 4, 5, 6, 7, or 8. X+< represents a monovalent cation or the nth part of an n-valent cation, preferably the alkali metal ions, including Na+< or K+<, with Na+< being highly preferred. Further cations X+< can be selected from NH4+<, ½ Zn2+<, ½ Mg2+<, ½ Ca2+<, ½ Mn2+<, and mixtures thereof.

[0078] In various embodiments, the alkyl ether sulfate can be selected from fatty alcohol ether sulfates of the formula with k = 11 to 19, n = 2, 3, 4, 5, 6, 7, or 8. Particularly preferred representatives are Na-C 12-14 fatty alcohol ether sulfates with 2 EO (k = 11-13, n = 2). The stated degree of ethoxylation represents a statistical average, which may be a whole number or a fraction for a specific product. The stated degrees of alkoxylation represent statistical averages, which may be a whole number or a fraction for a specific product. Preferred alkoxylates / ethoxylates exhibit a narrow range of homologs (nre ethoxylates).

[0079] For cold washing performance, it has proven advantageous for detergents to contain additional soap(s). Preferred detergents are therefore characterized by the fact that they contain soap(s). Suitable soaps include saturated fatty acid soaps, such as the salts of lauric acid, myristic acid, palmitic acid, stearic acid, hydrogenated erucic acid, and behenic acid, as well as, in particular, soap mixtures derived from natural fatty acids, e.g., coconut, palm kernel, or tallow fatty acids.

[0080] Suitable nonionic surfactants are, in particular, alkyl glycosides and ethoxylation and / or propoxylation products of alkyl glycosides or linear or branched alcohols, each with 8 to approximately 18 carbon atoms in the alkyl moiety and 3 to 20, preferably 4 to 10, alkyl ether groups. Furthermore, corresponding ethoxylation and / or propoxylation products of N-alkylamines, vicinal diols, fatty acid esters, and fatty acid amides, which correspond to the aforementioned long-chain alcohol derivatives with respect to the alkyl moiety, as well as of alkylphenols with 5 to 12 carbon atoms in the alkyl group, are also suitable.

[0081] Preferably, alkoxylated, advantageously ethoxylated, and especially primary alcohols with preferably 8 to 18 carbon atoms and an average of 1 to 12 moles of ethylene oxide (EO) per mole of alcohol are used as nonionic surfactants. The alcohol residue in these alcohols may be linear or, preferably, methyl-branched at the 2-position, or the mixture may contain both linear and methyl-branched residues, as is commonly found in oxo alcohol residues. However, alcohol ethoxylates with linear residues derived from native alcohols with 12 to 18 carbon atoms, e.g., from coconut, palm, tallow, or oleyl alcohol, and an average of 2 to 8 EO per mole of alcohol, are particularly preferred. Preferred ethoxylated alcohols include, for example, C 12-14 alcohols with 3 EO or 4 EO, C 9-11 alcohol with 7 EO, C 13-15 alcohols with 3 EO, 5 EO, 7 EO or 8 EO, C 12-18 alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of C 12-14 alcohol with 3 EO and C 12-18 alcohol with 5 EO.The stated degrees of ethoxylation represent statistical averages, which may be whole numbers or fractions for a specific product. Preferred alcohol ethoxylates exhibit a narrow range of homologs (NRE). In addition to these nonionic surfactants, fatty alcohols with more than 12 EO can also be used. Examples include tallow fatty alcohols with 14 EO, 25 EO, 30 EO, or 40 EO.

[0082] Another class of preferably used non-ionic surfactants, which are used either as the sole non-ionic surfactant or in combination with other non-ionic surfactants, are alkoxylated, preferably ethoxylated or ethoxylated and propoxylated fatty acid alkyl esters, preferably with 1 to 4 carbon atoms in the alkyl chain, in particular fatty acid methyl esters.

[0083] Another class of nonionic surfactants that can be used advantageously are the alkyl polyglycosides (APGs). Suitable alkyl polyglycosides conform to the general formula RO(G)z, where R represents a linear or branched, particularly 2-methyl-branched, saturated or unsaturated, aliphatic residue with 8 to 22, preferably 12 to 18, carbon atoms, and G is the symbol representing a glucose unit with 5 or 6 carbon atoms, preferably glucose. The degree of glycosylation z is between 1.0 and 4.0, preferably between 1.0 and 2.0, and particularly between 1.1 and 1.4. Linear alkyl polyglycosides are preferred, i.e., alkyl polyglycosides in which the polyglycosyl residue is a glucose residue and the alkyl residue is an n-alkyl residue.

[0084] Non-ionic surfactants of the amine oxide type, e.g., N-cocosalkyl-N,N-dimethylamine oxide and N-tallowalkyl-N,N-dihydroxyethylamine oxide, and fatty acid alkanolamides may also be suitable. The amount of these non-ionic surfactants is preferably no more than that of the ethoxylated fatty alcohols, and in particular not more than half of it.

[0085] Non-ionic surfactants preferably used in the detergents according to the invention have the formula R 1< -CH(OH)CH 2 O-(AO) w -(A'O) x -(A"O) y -(A‴O) z -R 2<, in which R 1< represents a straight-chain or branched, saturated or mono- or polyunsaturated C 6-24 alkyl or -alkenyl residue; R 2< represents a linear or branched hydrocarbon residue with 2 to 26 carbon atoms; A, A', A" and A‴ independently represent a residue from the group -CH 2 CH 2 , -CH 2 CH 2 -CH 2 , -CH 2 -CH(CH 3 ), -CH 2 -CH 2 -CH 2 -CH 2 , -CH 2 -CH(CH 3 )-CH 2 -, -CH 2 -CH(CH 2 -CH 3 ) stand; w, x, y and z stand for values ​​between 0.5 and 120, where x, y and / or z can also be 0, are preferred.By adding the aforementioned non-ionic surfactants of the formula R 1< -CH(OH)CH 2 O-(AO) w -(A'O) x -(A"O) y -(A‴O) z -R 2< , hereinafter also referred to as "hydroxy mixed ether", the cleaning performance of enzyme-containing preparations according to the invention can be significantly improved, both in comparison to surfactant-free systems and in comparison to systems containing alternative non-ionic surfactants, e.g. from the group of polyalkoxylated fatty alcohols.

[0086] The use of these non-ionic surfactants with one or more free hydroxyl group(s) on one or both terminal alkyl groups can significantly improve the stability of the enzymes contained in the detergent preparations according to the invention.

[0087] Particularly preferred are end-group-capped poly(oxyalkylated) niotenes which, according to the formula R 1< O[CH 2 CH 2 O] x CH 2 CH(OH)R 2< , in addition to a residue R 1< , which represents linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon residues with 2 to 30 carbon atoms, preferably with 4 to 22 carbon atoms, further comprise a linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon residue R 2< with 1 to 30 carbon atoms, where x represents values ​​between 1 and 90, preferably values ​​between 30 and 80 and particularly values ​​between 30 and 60.

[0088] Particularly preferred are surfactants of the formula R1< O[CH2CH(CH3)O] x [CH2CH2O] y CH2CH(OH)R2< , where R1< represents a linear or branched aliphatic hydrocarbon residue with 4 to 18 carbon atoms or mixtures thereof, R2< represents a linear or branched hydrocarbon residue with 2 to 26 carbon atoms or mixtures thereof, and x represents values ​​between 0.5 and 1.5 and y represents a value of at least 15. Examples of these nonionic surfactants include the C2-26 fatty alcohol (PO)1-(EO)15-40-2-hydroxyalkyl ethers, and in particular the C8-10 fatty alcohol (PO)1-(EO)22-2-hydroxydecyl ethers.

[0089] Particularly preferred are end-group-capped poly(oxyalkylated) niotenes of the formula R 1< O[CH 2 CH 2 O] x [CH 2 CH(R 3< )O] y CH 2 CH(OH)R 2< , in which R 1< and R 2< independently represent a linear or branched, saturated or mono- or polyunsaturated hydrocarbon residue with 2 to 26 carbon atoms, R 3< is independently selected from -CH 3 , -CH 2 CH 3 , -CH 2 CH 2 -CH 3 , -CH(CH 3 ) 2 , preferably, however, represents -CH 3, and x and y independently represent values ​​between 1 and 32, wherein niotenes with R 3< = -CH 3 and values ​​for x from 15 to 32 and y from 0.5 and 1.5 are particularly preferred.

[0090] Other preferably usable niotenes are the end-capped poly(oxyalkylated) niotenes of the formula R 1< O[CH 2 CH(R 3< )O] x [CH 2 ] k CH(OH)[CH 2 ] j OR 2< , in which R 1< and R 2< represent linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon residues with 1 to 30 carbon atoms, R 3< represents H or a methyl, ethyl, n-propyl, iso-propyl, n-butyl, 2-butyl or 2-methyl-2-butyl residue, x represents values ​​between 1 and 30, k and j represent values ​​between 1 and 12, preferably between 1 and 5. If the value x ≥ 2, each R 3< in the formula above R 1< O[CH 2 CH(R 3< )O] x [CH 2 ] k CH(OH)[CH 2 ] j OR 2< can be different. R 1< and R 2< are preferably linear or branched, saturated or unsaturated, aliphatic or aromatic hydrocarbon residues with 6 to 22 carbon atoms, with residues having 8 to 18 carbon atoms being particularly preferred.For the residue R3<, H, -CH3, or -CH2CH3 are particularly preferred. Particularly preferred values ​​for x are in the range of 1 to 20, especially 6 to 15. As described above, each R3< in the formula above can be different if x ≥ 2. This allows the alkylene oxide unit in the square brackets to be varied. For example, if x represents 3, the residue R3< can be selected to form ethylene oxide (R3< = H) or propylene oxide (R3< = CH3) units, which can be joined together in any order, e.g., (EO)(PO)(EO), (EO)(EO)(PO), (EO)(EO)(EO), (PO)(EO)(PO), (PO)(PO)(EO), and (PO)(PO)(PO). The value 3 for x has been chosen here as an example and can certainly be larger, whereby the range of variation increases with increasing x-values ​​and includes, for example, a large number of (EO) groups combined with a small number of (PO) groups, or vice versa.Particularly preferred end-capped poly(oxyalkylated) alcohols of the formula above have values ​​of k = 1 and j = 1, so that the above formula simplifies to R1< O[CH2CH(R3< )O] x CH2CH(OH)CH2OR2<. In the latter formula, R1<, R2<, and R3< are defined as above, and x represents numbers from 1 to 30, preferably from 1 to 20, and particularly from 6 to 18. Surfactants in which the R1< and R2< groups have 9 to 14 carbon atoms, R3< represents hydrogen, and x takes on values ​​from 6 to 15 are particularly preferred.

[0091] Finally, the nonionic surfactants of the formula R 1< -CH(OH)CH 2 O-(AO) w -R 2< have proven to be particularly effective, in which R 1< represents a straight-chain or branched, saturated or mono- or polyunsaturated C 6-24 -alkyl or -alkenyl residue; R 2< represents a linear or branched hydrocarbon residue with 2 to 26 carbon atoms; A represents a residue from the group -CH 2 CH 2 , -CH 2 CH 2 -CH 2 , -CH 2 -CH(CH 3 ), and w represents values ​​between 1 and 120, preferably 10 to 80, in particular 20 to 40. Examples of these non-ionic surfactants include the C 4-22 fatty alcohol (EO) 10-80 2-hydroxyalkyl ethers, in particular the C 8-12 fatty alcohol (EO) 22 2-hydroxydecyl ethers and the C 4-22 fatty alcohol (EO) 40-80 2-hydroxyalkyl ethers.

[0092] Preferred detergents are characterized in that the detergent contains at least one non-ionic surfactant, preferably a non-ionic surfactant from the group of hydroxy mixed ethers, wherein the weight fraction of the non-ionic surfactant to the total weight of the detergent is preferably 0.2 to 20 wt.%, preferably 1 to 18 wt.%, more preferably 2 to 15 wt.% and particularly preferably 5 to 10 wt.%.

[0093] Suitable amphoteric surfactants are, for example, betaines of the formula (R iii< )(R iv< )(R v< )N +< CH 2 COO -< , in which R iii< represents an alkyl group with 8 to 25, preferably 10 to 21 carbon atoms, possibly interrupted by heteroatoms or heteroatom groups, and R iv< and R v< represent similar or different alkyl groups with 1 to 3 carbon atoms, in particular C 10-18 alkyldimethylcarboxymethyl betaine and C 11-17 alkylamidopropyldimethylcarboxymethyl betaine. Suitable cationic surfactants include, among others, the quaternary ammonium compounds of the formula (R vi< )(R vii< )(R viii< )(R ix< )N +< X -< , in which R vi< to R ix< represent four identical or different, in particular two long-chain and two short-chain, alkyl groups and X -< represent an anion, in particular a halide ion, e.g. didecyldimethylammonium chloride, alkylbenzyldidecylammonium chloride and their mixtures.Other suitable cationic surfactants are quaternary surfactants, particularly those with a sulfonium, phosphonium, iodonium, or arsonium group, which are also known as antimicrobial agents. By using quaternary surfactants with antimicrobial activity, the product can be formulated with antimicrobial properties, or its existing antimicrobial effect, if present due to other ingredients, can be enhanced.

[0094] Another preferred component of detergents according to the invention are complexing agents. Phosphonates are particularly preferred complexing agents, provided their use is permitted under regulations. In addition to 1-hydroxyethane-1,1-diphosphonic acid, the complexing phosphonates include a number of different compounds, such as diethylenetriaminepenta(methylenephosphonic acid) (DTPMP). In this application, hydroxyalkane and aminoalkane phosphonates are particularly preferred. Among the hydroxyalkane phosphonates, 1-hydroxyethane-1,1-diphosphonate (HEDP) is of particular importance as a cobuilder. It is preferably used as a sodium salt, with the disodium salt being neutral and the tetrasodium salt reacting alkaline (pH 9). Ethylenediaminetetramethylenephosphonate (EDTMP), diethylenetriaminepentamethylenephosphonate (DTPMP), and their higher homologs are preferably suitable as aminoalkane phosphonates. They are preferably used in the form of the neutral sodium salts, e.g.,Aminoalkane phosphonates are used as the hexasodium salt of EDTMP or as the hepta- and octa-sodium salts of DTPMP. HEDP, from the phosphonate class, is preferred as a builder. Furthermore, aminoalkane phosphonates possess a pronounced heavy metal binding capacity. Accordingly, especially when the products also contain bleach, it may be preferable to use aminoalkane phosphonates, particularly DTPMP, or mixtures of the aforementioned phosphonates.A detergent preferred in this application contains one or more phosphonates from the group consisting of aminotrimethylenephosphonic acid (ATMP) and / or its salts; ethylenediaminetetra(methylenephosphonic acid) (EDTMP) and / or its salts; diethylenetriaminepenta(methylenephosphonic acid) (DTPMP) and / or its salts; 1-hydroxyethane-1,1-diphosphonic acid (HEDP) and / or its salts; 2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC) and / or its salts; hexamethylenediaminetetra(methylenephosphonic acid) (HDTMP) and / or its salts; nitrilotri(methylenephosphonic acid) (NTMP) and / or its salts. Detergents containing 1-hydroxyethane-1,1-diphosphonic acid (HEDP) or diethylenetriaminepenta(methylenephosphonic acid) (DTPMP) as phosphonates are particularly preferred. Naturally, the detergents according to the invention can contain two or more different phosphonates.Preferred detergents according to the invention are characterized in that the detergent contains at least one complexing agent from the group of phosphonates, preferably 1-hydroxyethane-1,1-diphosphonate, wherein the weight fraction of the phosphonate to the total weight of the detergent is preferably 0.1 and 8.0 wt.%, more preferably 0.2 and 5.0 wt.%, further preferably 0.3 and 3.0 wt.% and particularly preferably 0.5-2.0 wt.%.

[0095] The detergents according to the invention further preferably contain a builder, preferably at least one water-soluble and / or water-insoluble, organic and / or inorganic builder. The builders include, in particular, silicates, carbonates, and organic co-builders.

[0096] Examples of organic cobuilders include polycarboxylates / polycarboxylic acids, polymeric polycarboxylates, aspartic acid, polyacetals, dextrins, other organic cobuilders, and phosphonates. These classes of substances are described below. Organic cobuilder substances can be present, if desired, in amounts up to 40% by weight, particularly up to 25% by weight, and preferably from 1 to 8% by weight.

[0097] Useful organic framework substances include, for example, polycarboxylic acids which can be used in the form of the free acid and / or their sodium salts, whereby polycarboxylic acids are understood to be carboxylic acids that carry more than one acid function.Examples include citric acid, adipic acid, succinic acid, glutaric acid, malic acid, tartaric acid, maleic acid, fumaric acid, sugar acids and carboxymethylinulins, monomeric and polymeric aminopolycarboxylic acids, in particular glycine diacetic acid, methylglycine diacetic acid, glutamine diacetic acid, nitrilotriacetic acid (NTA), iminodisuccinates such as ethylenediamine-N,N'-disuccinic acid and hydroxyiminodisuccinates, ethylenediaminetetraacetic acid and polyaspartic acid, polyphosphonic acids, in particular aminotris(methylenephosphonic acid), ethylenediaminetetrakis(methylenephosphonic acid), lysinetetra(methylenephosphonic acid) and 1-hydroxyethane-1,1-diphosphonic acid, polymeric hydroxy compounds such as dextrin and polymeric (poly)carboxylic acids, in particular by oxidation of polysaccharides or...Dextrin-accessible polycarboxylates, and / or polymeric acrylic acids, methacrylic acids, maleic acids, and copolymers thereof, which may also contain small amounts of polymerizable substances without carboxylic acid functionality. Such organic builder substances may, if desired, be present in amounts up to 50 wt.%, in particular up to 25 wt.%, preferably from 10 to 20 wt.%, and most preferably from 1 to 5 wt.%.

[0098] In addition to their builder properties, free acids typically also possess the properties of an acidifying component and thus serve to adjust detergents to a lower and milder pH value. Citric acid, succinic acid, glutaric acid, adipic acid, gluconic acid, and any mixtures thereof are particularly noteworthy in this regard. Citric acid or its salts are especially preferred as the building block. Other particularly preferred building blocks include methylglycine diside acid (MGDA), glutamic acid diacetate (GLDA), aspartic acid diacetate (ASDA), hydroxyethyliminodiacetate (HEIDA), iminodisuccinate (IDS), ethylenediamine disuccinate (EDDS), carboxymethylinulin, and polyaspartate.

[0099] In preferred embodiments, citric acid and / or citrate are used as water-soluble, organic builders. Particularly preferred is the use of 0.5 to 25 wt.%, preferably 0.75 to 12.5 wt.%, more preferably 1 to 4 wt.% citric acid and / or 0.5 to 25 wt.%, preferably 0.75 to 12.5 wt.%, more preferably 1 to 4 wt.% citrate, preferably alkali citrate, and even more preferably sodium citrate. Citric acid and citrate can each be used in the form of their hydrates; for example, citric acid can be used in the form of the monohydrate, and citrate in the form of the trisodium citrate dihydrate.

[0100] Suitable structural materials also include polymeric polycarboxylates, such as the alkali metal salts of polyacrylic acid or polymethacrylic acid, e.g., those with a relative molecular mass of 500 to 70,000 g / mol. For the purposes of this application, the molar masses specified for polymeric polycarboxylates are weight-average molar masses Mw of the respective acid form, which were generally determined by gel permeation chromatography (GPC) using a UV detector. The measurement was performed against an external polyacrylic acid standard, which, due to its structural similarity to the polymers under investigation, provides realistic molar mass values. These values ​​differ significantly from the molar mass values ​​obtained when polystyrenesulfonic acids are used as the standard. The molar masses measured against polystyrenesulfonic acids are generally considerably higher than the molar masses specified in this application.Suitable polymers are, in particular, polyacrylates, preferably having a molecular weight of 2,000 to 20,000 g / mol. Due to their superior solubility, short-chain polyacrylates from this group, with molecular weights of 2,000 to 10,000 g / mol, and especially preferably 3,000 to 5,000 g / mol, are particularly preferred. Copolymers of polycarboxylates are also suitable, especially those of acrylic acid with methacrylic acid and of acrylic acid or methacrylic acid with maleic acid. Copolymers of acrylic acid with maleic acid containing 50 to 90 wt% acrylic acid and 50 to 10 wt% maleic acid have proven to be particularly suitable. Their relative molecular mass, based on free acids, is generally 2,000 to 70,000 g / mol, preferably 20,000 to 50,000 g / mol and particularly 30,000 to 40,000 g / mol.

[0101] A solid composition according to the invention preferably contains at least one water-soluble and / or water-insoluble, organic and / or inorganic builder. The water-soluble organic builder substances include the organic scaffold substances mentioned above.

[0102] In addition to the aforementioned water-soluble organic builders, the compositions of the invention may also contain inorganic water-soluble builders. Suitable water-soluble inorganic builder materials include, in particular, alkali silicates, alkali carbonates, alkali hydrogen carbonates, alkali phosphates, and / or sesquicarbonates, which may be present in the form of their alkaline, neutral, or acidic sodium or potassium salts. Small amounts of calcium carbonate may also be present in solid textile detergents. Suitable examples include water-soluble crystalline and / or amorphous alkali silicates. The alkali silicates suitable as builders in the compositions of the invention preferably have a molar ratio of alkali oxide to SiO₂ below 0.95, particularly from 1:1.1 to 1:12, and may be amorphous or crystalline.Preferred alkali silicates are sodium silicates, particularly amorphous sodium silicates, with a molar ratio Na₂O:SiO₂ of 1:2 to 1:2.8. Crystalline silicates, which may be present alone or in mixtures with amorphous silicates, are preferably crystalline layered silicates of the general formula Na₂Si₆O₆ 2x+1 · y H₂O, where x, the so-called modulus, is a number from 1.9 to 22, particularly 1.9 to 4, and y is a number from 0 to 33, with preferred values ​​for x being 2, 3, or 4. Preferred crystalline layered silicates are those in which x in the aforementioned general formula takes the values ​​2 or 3. In particular, both β- and δ-sodium disilicates (Na₂Si₂O₅ y H₂O) are preferred. Practically anhydrous crystalline alkali silicates of the above-mentioned general formula, in which x represents a number from 1.9 to 2.1, produced from amorphous alkali silicates, can also be used in compositions according to the invention.In a further embodiment of the composition according to the invention, a crystalline sodium layered silicate with a modulus of 2 to 3 is used, such as can be produced from sand and soda. Crystalline sodium silicates with a modulus in the range of 1.9 to 3.5 are used in a further embodiment of the composition according to the invention. In compositions containing both amorphous and crystalline alkali silicates, the weight ratio of amorphous alkali silicate to crystalline alkali silicate is preferably 1:2 to 2:1 and particularly 1:1 to 2:1. Crystalline layered silicates of the above-mentioned formula (I) are marketed by Clariant GmbH under the trade name Na-SKS, e.g. Na-SKS-1 (Na 2 Si 22 O 45 x H 2 O, Kenyaite), Na-SKS-2 (Na 2 Si 14 O 29 x H 2 O, Magadiite), Na-SKS-3 (Na 2 Si 8 O 17 x H 2 O) or Na-SKS-4 (Na 2 Si 4 O 9 x H 2 O, Makatite).Of these, Na-SKS-5 (α-Na₂Si₂O₅), Na-SKS-7 (β-Na₂Si₂O₅, natrosilite), Na-SKS-9 (NaHSi₂O₅₃H₂O), Na-SKS-10 (NaHSi₂O₅₃H₂O, kanemite), Na-SKS-11 (t-Na₂Si₂O₅) and Na-SKS-13 (NaHSi₂O₅), but especially Na-SKS-6 (δ-Na₂Si₂O₅), are particularly suitable. In one embodiment of the inventive composition, a granular compound of crystalline layered silicate and citrate, of crystalline layered silicate and the aforementioned (co-)polymeric polycarboxylic acid, or of alkali silicate and alkali carbonate is used, such as is commercially available under the name Nabion®< 15. Such water-soluble inorganic builder materials are preferably contained in the compositions according to the invention in amounts of 1 to 20 wt.%, in particular 5 to 15 wt.%. Furthermore, carbonates (and hydrogen carbonates), in particular sodium carbonate, and phosphonic acids / phosphonates are also important as water-soluble inorganic builder substances.The term phosphonic acids also includes, where applicable, substituted alkylphosphonic acids, which may also contain multiple phosphonic acid groups (so-called polyphosphonic acids). They are preferably selected from the hydroxy and / or aminoalkylphosphonic acids and / or their alkali salts, such as dimethylaminomethanediphosphonic acid, 3-aminopropane-1-hydroxy-1,1-diphosphonic acid, 1-amino-1-phenylmethanediphosphonic acid, 1-hydroxyethane-1,1-diphosphonic acid (HEDP), aminotris-(methylenephosphonic acid), N,N,N',N'-ethylenediaminetetrakis-(methylenephosphonic acid), diethylenetriaminepenta-(methylenephosphonic acid) (DTPMP), and acylated derivatives of phosphorous acids, which can also be used in any mixtures.

[0103] In various embodiments, the builder system preferably consists of the following components, each based on the total mass of the medium: a) 0 to 10 wt.%, preferably 1 to 4 wt.% citric acid; and / or b) 0 to 10 wt.%, preferably 1 to 4 wt.% citrate, preferably alkali citrate; and / or c) 0 to 40 wt.%, preferably 0 to 15 wt.%, particularly preferably 1 to 3 wt.% alkali carbonate, which may also be at least partially replaced by alkali hydrogen carbonate, in particular sodium carbonate; and / or d) 0 to 20 wt.%, preferably 3 to 10 wt.% alkali silicate; e) 0 to 10 wt.%, preferably 0.02 to 2 wt.%, particularly preferably 0.5 to 2 wt.% phosphonic acid and / or alkali phosphonate, in particular HEDP and / or DTPMP; and / or f) 0 to 10 wt.%, preferably 0.5 to 3 wt.% polymeric polycarboxylate, in particular polyacrylate.

[0104] Regarding component f), suitable materials include, for example, the alkali metal salts of polyacrylic acid or polymethacrylic acid, such as those with a relative molecular mass of 500 to 70,000 g / mol. This class of substances has already been described in detail above. The (co-)polymeric polycarboxylates can be used either as a powder or as an aqueous solution.

[0105] The compositions according to the invention are preferably free of phosphate builders, i.e., they contain less than 1 wt% and preferably no deliberately added phosphate builder.

[0106] The agents may also contain water-insoluble builder substances. Particularly suitable water-insoluble inorganic builder materials are crystalline or amorphous water-dispersible alkali aluminosilicates, in amounts of up to 50 wt.%, preferably not exceeding 40 wt.%, particularly 3 to 20 wt.%, and most preferably 1 to 15 wt.%. Among these, crystalline sodium aluminosilicates of detergent quality, especially zeolite A, zeolite P, zeolite MAP, and optionally zeolite X, alone or in mixtures, e.g., in the form of a co-crystallizate of zeolites A and X (Vegobond®<AX, a commercial product of Condea Augusta SpA), are preferred. Amounts close to the aforementioned upper limit are preferably used in solid, particulate agents. Suitable aluminosilicates, in particular, do not contain particles with a grain size greater than 30 µm and preferably consist of at least 80 wt.% of particles with a size less than 10 µm.Their calcium binding capacity, which can be determined according to DE 2412837 A1, is usually in the range of 100 to 200 mg CaO per gram.

[0107] In addition to the previously described base materials, the detergent may contain cleaning-active polymers. The weight fraction of the cleaning-active polymers in the total weight of the detergent according to the invention is preferably 0.1 to 20 wt.%, more preferably 1.0 to 15 wt.%, and more preferably 2.0 to 12 wt.%. Preferably, polymers containing sulfonic acid groups, in particular from the group of copolymeric polysulfonates, are used as cleaning-active polymers. These copolymeric polysulfonates contain, in addition to sulfonic acid group-containing monomer(s), at least one monomer from the group of unsaturated carboxylic acids.Unsaturated carboxylic acids of the formula R 1< (R 2< )C=C(R 3< )COOH are particularly preferred, in which R 1< to R 3< independently represent -H, -CH 3 , a straight-chain or branched saturated alkyl group with 2 to 12 carbon atoms, a straight-chain or branched, mono- or poly-unsaturated alkenyl group with 2 to 12 carbon atoms, alkyl or alkenyl groups substituted with -NH 2 , -OH or -COOH as defined above, or -COOH or -COOR 4< , where R 4< is a saturated or unsaturated, straight-chain or branched hydrocarbon group with 1 to 12 carbon atoms. Particularly preferred unsaturated carboxylic acids are acrylic acid, methacrylic acid, ethacrylic acid, α-chloroacrylic acid, α-cyanoacrylic acid, crotonic acid, α-phenylacrylic acid, maleic acid, maleic anhydride, fumaric acid, itaconic acid, citraconic acid, methylenemalonic acid, sorbic acid, cinnamic acid or mixtures thereof.Unsaturated dicarboxylic acids can also be used, of course. For sulfonic acid group-containing monomers, those of the formula R 5< (R 6< )C=C(R 1< )-X-SO 3 H are preferred, in which R 5< to R 7< independently represent -H, -CH 3 , a straight-chain or branched saturated alkyl group with 2 to 12 carbon atoms, a straight-chain or branched, mono- or polyunsaturated alkenyl group with 2 to 12 carbon atoms, alkyl or alkenyl groups substituted with -NH 2 , -OH or -COOH, or -COOH or -COOR 4<, where R 4< is a saturated or unsaturated, straight-chain or branched hydrocarbon group with 1 to 12 carbon atoms, and X represents an optional spacer group selected from -(CH 2 ) n - with n = 0 to 4. -COO-(CH 2 ) k - with k = 1 to 6, -C(O)-NH-C(CH 3 ) 2 -, -C(O)-NH-C(CH 3 ) 2 -CH 2 - and -C(O)-NH-CH(CH 2 CH 3 )-.Among these monomers, preferred are those of the formulas H₂C=CH-X-SO₃H, H₂C=C(CH₃)-X-SO₃H, and HO₃SX-(R₆<)C=C(R₇<)-X-SO₃H, in which R₆< and R₇< are independently selected from -H, -CH₃, -CH₂CH₃, -CH₂CH₂CH₃, -CH(CH₃)₂, and X represents an optional spacer group selected from -(CH₂)ₙ - with n = 0 to 4, -COO-(CH₂)ₖ - with k = 1 to 6, -C(O)-NH-C(CH₃)₂ -, -C(O)-NH-C(CH₃)₂ -CH₂ -, and -C(O)-NH-CH(CH₂CH₃)₂ 3 )-.Particularly preferred monomers containing sulfonic acid groups include 1-acrylamido-1-propanesulfonic acid, 2-acrylamido-2-propanesulfonic acid, 2-acrylamido-2-methyl-1-propanesulfonic acid, 2-methacrylamido-2-methyl-1-propanesulfonic acid, 3-methacrylamido-2-hydroxy-propanesulfonic acid, allylsulfonic acid, metallylsulfonic acid, allyloxybenzenesulfonic acid, metallyloxybenzenesulfonic acid, 2-hydroxy-3-(2-propenyloxy)-propanesulfonic acid, 2-methyl-2-propenesulfonic acid, styrenesulfonic acid, vinylsulfonic acid, 3-sulfopropyl acrylate, 3-sulfopropyl methacrylate, sulfomethacrylamide, sulfomethylmethacrylamide, and mixtures of the aforementioned acids or their water-soluble salts. The sulfonic acid groups in the polymers may be present wholly or partially in neutralized form. The use of partially or fully neutralized copolymers containing sulfonic acid groups is preferred according to the invention.The molar mass of the sulfo copolymers preferably used according to the invention can be varied to adapt the properties of the polymers to the desired application. Preferably, monomers of the formula R1< (R2< )C=C(R3< )-XR4< are used as nonionic monomers, in which R1< to R3< independently represent -H, -CH3 or -C2H5, X represents an optional spacer group selected from -CH2-, -C(O)O- and -C(O)-NH-, and R4< represents a straight-chain or branched saturated alkyl group with 2 to 22 carbon atoms or an unsaturated, preferably aromatic, group with 6 to 22 carbon atoms.Particularly favored nonionic monomers are butene, isobutene, pentene, 3-methylbutene, 2-methylbutene, cyclopentene, hexene, hexene-1, 2-methylpentene-1, 3-methylpentene-1, cyclohexene, methylcyclopentene, cycloheptene, methylcyclohexene, 2,4,4-trimethylpentene-1, 2,4,4-trimethylpentene-2, 2,3-dimethylhexene-1, 2,4-diemethylhexene-1, 2,5-dimethylhexene-1, 3,5-dimethylhexene-1, 4,4-dimethylhexane-1, ethylcyclohexyne, 1-octene, and α-olefins with 10 or more carbon atoms, such as...1-Decene, 1-Dodecene, 1-Hexadecene, 1-Octadecene and C 22-α-olefin, 2-Styrene, α-Methylstyrene, 3-Methylstyrene, 4-Propylstyrene, 4-Cyclohexylstyrene, 4-Dodecylstyrene, 2-Ethyl-4-Benzylstyrene, 1-Vinylnaphthalene, 2-Vinylnaphthalene, Methyl acrylate, Ethyl acrylate, Propyl acrylate, Butyl acrylate, Pentyl acrylate, Hexyl acrylate, Methyl methacrylate, N-(Methyl)acrylamide, 2-Ethylhexyl acrylate, Methacrylic acid 2-Ethylhexyl acrylate, N-(2-Ethylhexyl)acrylamide, Octyl acrylate, Methacrylic acid octyl acrylate, N-(Octyl)acrylamide Acrylic lauryl esters, methacrylic lauryl esters, N-(lauryl)acrylamide, acrylic stearyl esters, methacrylic stearyl esters, N-(stearyl)acrylamide, acrylic behenyl esters, methacrylic behenyl esters, and N-(behenyl)acrylamide, or mixtures thereof. The weight fraction of the sulfonic acid group-containing copolymers in the total weight of the detergents according to the invention is preferably 0.1 to 15 wt.%, preferably 1.0 to 12 wt.%.-% and preferably 2.0 to 10 wt.%.

[0108] In one embodiment, a detergent comprises, 5 to 70 wt.%, in particular 5 to 30 wt.% surfactants and / or 0 to 10 wt.%, in particular 1 to 3 wt.% water-soluble or water-dispersible inorganic builder material and / or 0 to 10 wt.%, in particular 1 to 4 wt.%, water-soluble organic builder substances and / or 0.01 to 5 wt.% complexing agents and / or 0.01 to 5 wt.% graying inhibitor and / or 0.01 to 5 wt.% foaming inhibitor, and / or 0.01 to 5 wt.% optical brighteners.

[0109] In a preferred embodiment, a liquid detergent comprises, 5 to 20 wt.%, in particular 9 to 17 wt.% anionic surfactants and / or 1 to 20 wt.%, in particular 5 to 9 wt.% non-ionic surfactants and / or 0.5 to 10 wt.%, in particular 1 to 4 wt.%, water-soluble organic builder substances, in particular citric acid or citrate, and / or 0.01 to 5 wt.%, in particular 0.5 to 2 wt.% complexing agents, in particular HEDP, and / or 0.01 to 5 wt.%, in particular 0.2 to 1 wt.% graying inhibitor and / or 0.01 to 5 wt.% foam inhibitor.

[0110] In another preferred embodiment, a liquid detergent comprises: 0 to 10 wt.%, in particular 1 to 3 wt.% anionic surfactants and / or 5 to 20 wt.%, in particular 10 to 18 wt.% non-ionic surfactants and / or 1 to 10 wt.%, in particular 1.5 to 3 wt.% water-soluble or water-dispersible inorganic builder material and / or 0 to 1 wt.%, in particular 0.01 to 0.5 wt.% perfumes and / or 0.01 to 5 wt.%, in particular 0.05 to 0.5 wt.% optical brighteners.

[0111] Suitable anti-graying agents or soil-release agents (soil-release polymers) are cellulose ethers, such as carboxymethylcellulose, methylcellulose, hydroxyalkylcelluloses, and cellulose mixtures, such as methylhydroxyethylcellulose, methylhydroxypropylcellulose, and methyl carboxymethylcellulose. Sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and mixtures thereof, and optionally mixtures thereof with methylcellulose, are preferably used. Commonly used soil-release agents include copolyesters containing dicarboxylic acid units, alkylene glycol units, and polyalkylene glycol units. The proportion of anti-graying agents and / or soil-release agents in compositions according to the invention generally does not exceed 2% by weight and is preferably 0.5 to 1.5% by weight, particularly preferably 0.5 to 2% by weight.

[0112] Optical brighteners, particularly for textiles made from cellulose fibers (e.g., cotton), can include derivatives of diaminostilbene disulfonic acid or its alkali metal salts. Suitable examples are salts of 4,4'-bis(2-anilino-4-morpholino-1,3,5-triazin-6-yl-amino)stilbene-2,2'-disulfonic acid or similarly structured compounds that, instead of the morpholino group, contain a diethanolamino group, a methylamino group, or a 2-methoxyethylamino group. Brighteners of the type of substituted 4,4'-distyryldiphenyl, such as 4,4'-bis-(4-chloro-3-sulfostyryl)diphenyl, may also be present. Mixtures of brighteners can also be used. Optical brighteners of the 1,3-diaryl-2-pyrazoline type, e.g., 1-(p-sulfoamoylphenyl)-3-(p-chlorophenyl)-2-pyrazolin, and similarly structured compounds, are particularly suitable for polyamide fibers. The content of the optical brightener or brightener mixture is generally not higher than 1% by weight, preferably 0.05 to 0.5% by weight.-%. In a preferred embodiment of the invention, the agent is free of such active ingredients.

[0113] Conventional defoamers that can be used in the compositions according to the invention include, for example, polysiloxane-silica mixtures, wherein the finely divided silica contained therein is preferably silanized or otherwise hydrophobized. The polysiloxanes can consist of linear compounds as well as cross-linked polysiloxane resins and mixtures thereof. Other defoamers include paraffin hydrocarbons, in particular microparaffins and paraffin waxes, whose melting point is above 40°C, saturated fatty acids or soaps with, in particular, 20 to 22 carbon atoms, e.g., sodium behenate, and alkali salts of phosphoric acid mono- and / or dialkyl esters in which the alkyl chains each have 12 to 22 carbon atoms. Among these, sodium monoalkyl phosphate and / or dialkyl phosphate with C 16-18 alkyl groups is preferably used. The proportion of the defoamers is preferably 0.2 to 2 wt.%, and particularly preferably not more than 1 wt.%.

[0114] To adjust the desired pH value, the compositions according to the invention can contain system- and environmentally compatible acids, in particular citric acid, acetic acid, tartaric acid, malic acid, lactic acid, glycolic acid, succinic acid, glutaric acid and / or adipic acid, but also mineral acids, in particular sulfuric acid or alkali hydrogen sulfates, or bases, in particular ammonium or alkali hydroxides, preferably sodium hydroxide. Such pH regulators are preferably not present in the compositions according to the invention in amounts exceeding 10% by weight, in particular from 0.5 to 6% by weight, and most preferably from 0.3 to 2% by weight.

[0115] Suitable peroxygen compounds for use in compositions according to the invention include, in particular, organic peracids or pericy salts of organic acids, such as phthalimidopercaproic acid, perbenzoic acid, or salts of diperdodecanedioic acid, hydrogen peroxide, and inorganic salts that release hydrogen peroxide under washing conditions, including perborate, percarbonate, persilicate, and / or persulfate such as caroate, as well as hydrogen peroxide inclusion compounds, such as H₂O₂-urea adducts. Hydrogen peroxide can also be generated using an enzymatic system, i.e., an oxidase and its substrate. If solid peroxygen compounds are to be used, these can be in the form of powders or granules, which can also be coated in a manner known in principle.The peroxygen compounds can be added to the washing liquor either as such or in the form of agents containing them, which in principle can contain all conventional washing, cleaning, or disinfecting agent components. Alkali percarbonate or alkali perborate monohydrate is particularly preferred. If an agent according to the invention contains peroxygen compounds, these are present in amounts preferably up to 50 wt.%, particularly from 5 to 30 wt.%, and more preferably from 0.1 to 20 wt.%.

[0116] The bleaching activators used in these agents can be compounds that, under perhydrolysis conditions, yield aliphatic peroxocarboxylic acids with preferably 1 to 10 carbon atoms, particularly 2 to 4 carbon atoms, and / or optionally substituted perbenzoic acid. Suitable substances include those bearing O- and / or N-acyl groups of the specified number of carbon atoms and / or optionally substituted benzoyl groups. Preferred are multiply acylated alkylenediamines, in particular tetraacetylethylenediamine (TAED), acylated triazine derivatives, in particular 1,5-diacetyl-2,4-dioxohexahydro-1,3,5-triazine (DADHT), acylated glycolurils, in particular tetraacetylglycoluril (TAGU), N-acylides, in particular N-nonanoylsuccinimide (NOSI), acylated phenolsulfonates or carboxylates or the sulfonic or carboxylic acids thereof, in particular nonanoyl- or isononanoyloxybenzenesulfonate or laroyloxybenzenesulfonate (NOBS or iso-NOBS or laroyloxybenzenesulfonate).LOBS), 4-(2-Decanoyloxyethoxycarbonyloxy)benzenesulfonate (DECOBS) or decanoyloxybenzoate (DOBA), carboxylic anhydrides, in particular phthalic anhydride, acylated polyhydric alcohols, in particular triacetin, ethylene glycol diacetate, 2,5-diacetoxy-2,5-dihydrofuran and enol esters, as well as acetylated sorbitol and mannitol or their described mixtures (SORMAN), acylated sugar derivatives, in particular pentaacetylglucose (PAG), pentaacetylfructose, tetraacetylxylose and octaacetyllactose, acetylated, optionally N-alkylated glucamine and gluconolactone, N-acylated lactams, e.g. N-benzoylcaprolactam, nitriles from which perimidic acids are formed, in particular aminoacetonitrile derivatives with a quaternized nitrogen atom, and / or Oxygen-transferring sulfonimines and / or acylhydrazones. Hydrophilically substituted acylacetals and acyllactams are also preferred. Combinations of conventional bleaching activators can also be used.Such bleach activators may be present, particularly in the presence of the aforementioned hydrogen peroxide-supplying bleaching agents, in the usual quantity range, preferably in amounts of 0.5 to 10 wt.%, in particular 1 to 8 wt.%, based on the total agent, but are preferably completely absent when percarboxylic acid is used as the sole bleaching agent.

[0117] In addition to or instead of conventional bleaching activators, solid products may also contain sulfonimines and / or bleaching-enhancing transition metal salts or transition metal complexes as so-called bleaching catalysts.

[0118] As a further component, the detergents according to the invention can contain an organic solvent. The addition of organic solvents has a beneficial effect on the enzyme stability and the cleaning performance of these agents. Preferred organic solvents are from the group consisting of monohydric or polyhydric alcohols, alkanolamines, or glycol ethers.Preferably, the solvents are selected from ethanol, n- or i-propanol, butanol, glycol, propanediol, butanediol, glycerin, diglycylene glycol, propyldiglycylene glycol, butyldiglycylene glycol, hexylene glycol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether, ethylene glycol mono-butyl ether, diethylene glycol methyl ether, diethylene glycol ethyl ether, propylene glycol methyl ether, propylene glycol ethyl ether, propylene glycol propyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, methoxytriglycol, ethoxytriglycol, butoxytriglycol, 1-butoxyethoxy-2-propanol, 3-methyl-3-methoxybutanol, propylene glycol t-butyl ether, and mixtures of these solvents. The weight fraction of these organic solvents in the total weight of the detergents according to the invention is preferably 0.1 to 10 wt.%, preferably 0.2 to 8.0 wt.% and further preferably 0.5 to 5.0 wt.%.A particularly preferred organic solvent, especially effective for stabilizing detergents, is glycerin and 1,2-propylene glycol. Liquid detergents preferably comprise at least one polyol, preferably from the group consisting of glycerin and 1,2-propylene glycol, based on the total weight of the detergent, preferably in amounts of 0.1 to 10 wt.%, more preferably 0.2 to 8.0 wt.%, and further preferably 0.5 to 5.0 wt.%. Other preferred organic solvents are organic amines and alkanolamines. The detergents according to the invention preferably contain these amines in amounts of 0.1 to 10 wt.%, more preferably 0.2 to 8.0 wt.%, and further preferably 0.5 to 5.0 wt.%, each based on their total weight. A particularly preferred alkanolamine is ethanolamine.

[0119] Another preferred component of the detergents according to the invention is a sugar alcohol (alditol). The group of alditols comprises non-cyclic polyols of the formula HOCH₂[CH(OH)]nCH₂OH. Examples of alditols include mannitol, isomalt, lactitol, sorbitol, xylitol, threitol, erythritol, and arabitol. Sorbitol has proven to be particularly advantageous with regard to enzyme stability. The weight fraction of the sugar alcohol in the total weight of the detergent is preferably 1.0 to 10 wt.%, more preferably 2.0 to 8.0 wt.%, and further preferably 3.0 to 6.0 wt.%.

[0120] Detergents according to the invention can contain only one protease as defined herein. Alternatively, they can also contain other enzymes in a concentration suitable for the efficacy of the agent. A further embodiment of the invention thus comprises agents that further include one or more additional enzymes. Preferably used as additional enzymes are all enzymes that can exhibit catalytic activity in the agent according to the invention, in particular lipase, amylase, cellulase, hemicellulase, mannanase, tannase, xylanase, xanthanase, xyloglucanase, β-glucosidase, pectinase, carrageenase, perhydrolase, oxidase, oxidoreductase, or other proteases—distinguishable from the proteases according to the invention—as well as mixtures thereof. Advantageously, the additional enzymes are each contained in the agent in an amount of 1 x 10⁻⁸ to 5 wt% based on active protein.Increasingly preferably, each further enzyme is contained in the composition according to the invention in an amount of 1 x 10⁻⁷ to 3 wt.%, from 0.00001 to 1 wt.%, from 0.00005 to 0.5 wt.%, from 0.0001 to 0.1 wt.%, and particularly preferably from 0.0001 to 0.05 wt.%, based on active protein. The enzymes particularly preferably exhibit synergistic cleaning performance against certain soils or stains, i.e., the enzymes contained in the composition mutually support each other in their cleaning performance. Such synergism is most preferably present between the protease contained in the composition according to the invention and another enzyme of a composition according to the invention, including, in particular, between the aforementioned protease and an amylase and / or a lipase and / or a mannanase and / or a cellulase and / or a pectinase.Synergistic effects can occur not only between different enzymes, but also between one or more enzymes and other ingredients of the composition according to the invention. Textile detergents preferred according to the invention comprise at least one protease and at least one amylase. In a further preferred embodiment of the invention, textile detergents comprise at least one protease and at least one cellulase. In a further preferred embodiment, textile detergents comprise at least one protease and at least one lipase. Textile detergents comprising 3 to 10 different enzymes are particularly preferred, with textile detergents comprising 3 to 10 different types of enzymes being particularly advantageous with regard to cleaning performance against a very wide range of stains.

[0121] Examples of proteases are the subtilisins BPN' from Bacillus amyloliquefaciens and Carlsberg Bacillus licheniformis,the protease PB92, the subtilisins 147 and 309, the protease from Bacillus lentus, subtilisin DY and the enzymes thermitase, proteinase K, and the proteases TW3 and TW7, which are classified as subtilases but no longer as subtilisins in the strict sense. Subtilisin Carlsberg is available in an advanced form under the trade name Alcalase® from Novozymes. Subtilisins 147 and 309 are marketed under the trade names Esperase® and Savinase®, respectively, by Novozymes. From the protease Bacillus lentusProtease variants are derived from DSM 5483. Other useful proteases include, for example, those marketed under the trade names Durazym®, Release®, Everlase®, Nafizym®, Natalase®, Kannase®, Progress Uno 101L®, and Ovozyme® by Novozymes; those marketed under the trade names Purafect®, Purafect® OxP, Purafect® Prime, Excellase®, Properase®, Preferenz P100®, and Preferenz P300® by Danisco / DuPont; those marketed under the trade name Lavergy pro 104 LS® by BASF; those marketed under the trade name Protosol® by Advanced Biochemicals Ltd.; those marketed under the trade name Wuxi® by Wuxi Snyder Bioproducts Ltd.; and those marketed under the trade names Proleather®. and Protease P® from Amano Pharmaceuticals Ltd., and the enzyme available under the name Proteinase K-16 from Kao Corp. Proteases from [missing information] are also particularly preferred. Bacillus gibsonii and Bacillus pumilus,which are disclosed in international patent applications WO 2008 / 086916 A1 and WO 2007 / 131656 A1. Further advantageously usable proteases are disclosed in patent applications WO 91 / 02792 A1, WO 2008 / 007319 A2, WO 93 / 18140 A1, WO 01 / 44452 A1, GB 1243784 A, WO 96 / 34946 A1, WO 02 / 029024 A1 and WO 03 / 057246 A1. Other usable proteases are those found in microorganisms Stenotrophomonas maltophilia, in particular Stenotrophomonas maltophilia K279a, Bacillus intermedius as well as Spherical Bacillus are naturally present.

[0122] Examples of amylases are the α-amylases from Bacillus licheniformis, Bacillus amyloliquefaciens or Bacillus stearothermophilus and especially their improved developments for use in detergents or cleaning agents. The enzyme from Bacillus licheniformisIt is available from Novozymes under the name Termamyl®< and from Danisco / DuPont under the name Purastar®< ST. Further developments of this α-amylase are available under the trade names Duramyl®< and Termamyl®< ultra (both from Novozymes), Purastar®< OxAm (Danisco / DuPont), and Keistase®< (Daiwa Seiko Inc.). The α-amylase of Bacillus amyloliquefaciens is marketed by the company Novozymes under the name BAN®, and derived variants of α-amylase from Bacillus stearothermophilus under the names BSG® and Novamyl®, also from the company Novozymes. Furthermore, α-amylase from [unclear text] is used for this purpose. Bacillus sp. A 7-7 (DSM 12368) and the cyclodextrin glucanotransferase (CGTase) from Bacillus agar-adherent(DSM 9948) is particularly noteworthy. Furthermore, the amylolytic enzymes disclosed in international patent applications WO 95 / 26397 A1, WO 96 / 23873 A1, WO 99 / 23211 A1, WO 00 / 60060 A2, WO 03 / 002711 A2, WO 03 / 054177 A2, WO 2006 / 002643 A2, WO 2007 / 079938 A2, WO 2011 / 100410 A2, and WO 2013 / 003659 A1 can be used. Fusion products of all the aforementioned molecules can also be used. In addition, the further developments of α-amylase available from Novozymes under the trade name Fungamyl® are also applicable. Aspergillus niger and A. riceSuitable. Other commercially available products that can be used advantageously include, for example, Amylase-LT® and Stainzyme® or Stainzyme® ultra or Stainzyme® plus, as well as Amplify™ 12L or Amplify Prime™ 100L, the latter also from Novozymes, and the PREFERENZ S® series from Danisco / DuPont, comprising, for example, PREFERENZ S100®, PREFERENZ S1000®, or PREFERENZ S210®. Variants of these enzymes obtained through point mutations can also be used according to the invention.

[0123] Preferred amylases comprise a) an α-amylase comprising an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:2 over its total length to at least 80%, increasingly preferably to at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and has at least one amino acid substitution at one of positions 172, 202, 208, 255 and 261 in the numbering according to SEQ ID NO:2, preferably selected from M202L, M202V, M202S, M202T, M202I, M202Q, M202W, The group consisting of S255N, R172Q and combinations thereof. The amylase particularly preferably comprises the amino acid substitution M202L or M202T.

[0124] Further preferred amylases comprise b) an α-amylase comprising an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:3 over its total length to at least 60%, increasingly preferably to at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and has at least one amino acid substitution at one of positions 9, 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 195, 202, 203, 214, 231, 256, 257, 258, 269, 270, 272, 283, 295, 296, 298, 299, 303, 304, 305, 311, 314, 315, 318, 319, 320, 323, 339, 345, 361, 378, 383, 419, 421, 437, 441, 444, 445, 446, 447, 450, 458, 461, 471, 482 and 484 and / or a deletion at any of the positions 183 and 184 in the count according to SEQ ID NO:3.Particularly preferred amylases have at least one amino acid substitution at one of positions 9, 26, 149, 182, 186, 202, 257, 295, 299, 323, 339 and 345, and / or particularly preferably at least one amino acid substitution or deletion selected from the group consisting of R118K, D183*, G184*, N195F, R320K, R458K and combinations thereof. Particularly preferred amylases comprise an amino acid sequence that is identical to the amino acid sequence specified in SEQ ID NO:3 over its entire length to at least 60%, increasingly preferably to at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and exhibit one of the following amino acid substitution / deletion combinations: . (i) M9L+M323T; (ii) M9L+M202L / T / V / I+M323T; (iii) M9L+N195F+M202L / T / V / I+M323T; (iv) M9L+R118K+D183*+G184*+R320K+M323T+R458K; (v) M9L+R118K+D183*+G184*+M202L / T / V / I+R320K+M323T+R458K; (vi) M9L+G149A+G182T+G186A+M202L+T2571+Y295F+N299Y+M323T+A339S+E345R; (vii) M9L+G149A+G182T+G186A+M2021+T257I+Y295F+N299Y+M323T+A339S+E345R; (viii) M9L+R118K+G149A+G182T+D183*+G184*+G1186A+M202L+T257I+Y295F+N299Y+R320K +M323T+A339S+E345R+R458K; (ix) M9L+R118K+G149A+G182T+D183*+G184*+G1186A+M202I+T257I+Y295F+N299Y+R320K +M323T+A339S+E345R+R458K; (x) M9L+R118K+D183*+D184*+N195F+M202L+R320K+M323T+R458K; (xi) M9L+R118K+D183*+D184*+N195F+M202T+R320K+M323T+R458K; (xii) M9L+R118K+D183*+D184*+N195F+M202I+R320K+M323T+R458K; (xiii) M9L+R118K+D183*+D184*+N195F+M202V+R320K+M323T+R458K; (xiv) M9L+R118K+N150H+D183*+D184*+N195F+M202L+V214T+R320K+M323T+R458K; (xv) M9L+R118K+D183*+D184*+N195F+M202L+V214T+R320K+M323T+E345N+R458K.

[0125] Further preferred amylases include c) an α-amylase comprising an amino acid sequence that is at least 90% identical over its total length to the amino acid sequence specified in SEQ ID NO:4 and optionally includes at least one substitution and / or deletion at one of positions 93, 116, 118, 129, 133, 134, 140, 142, 146, 147, 149, 151, 152, 169, 174, 183, 184, 186, 189, 193, 195, 197, 198, 200, 203, 206, 210, 212, 213, 235, 243, 244, 260, 262, 284, 303, 304, 320, 338, 347, 359, 418, 431, 434, 439, 447, 458, 469, 476 and 477 in the numbering according to SEQ ID NO:4, exhibits, preferably amino acid deletions at positions 183 and 184.Preferred amino acid substitutions include E260A / D / C / Q / L / M / F / P / S / W / V / G / H / I / K / N / R / T / Y, G304R / K / E / Q, W140Y / F, W189E / G / T, D134E, F262G / P, W284D / H / F / Y / R, W347H / F / Y, W439R / G, G476E / Q / R / K, G477E / Q / K / M / R, N195F / Y, N197F / L, Y198N, Y200F, Y203F, I206H / L / N / F / Y, H210Y, E212V / G, V213A, M116T, Q129L, G133E, E134Y, K142R, P146S, G147E, G149R, N151R, Y152H, Q169E, N174R, A186R, Y243F, S244Q, G303V, R320N, R359I, N418D and A447V.

[0126] Examples of cellulases (endoglucanases, EG) include the fungal, endoglucanase-rich cellulase preparation and its further developments, offered by Novozymes under the trade name Celluzyme®. The products Endolase® and Carezyme®, also available from Novozymes, are based on the 50 kDa EG and 43 kDa EG, respectively. Humicola insolensDSM 1800. Other usable commercial products from this company are Cellusoft®, Renozyme®, and Celluclean®. Also usable are, for example, cellulases available from AB Enzymes under the trade names Ecostone® and Biotouch®, which are at least partially based on the 20 kDa-EG standard. Melanocarpus based on. Other cellulases from AB Enzymes are Econase® and Ecopulp®. Other suitable cellulases are from Bacillus sp. CBS 670.93 and CBS 669.93, the latter consisting of Bacillus sp. CBS 670.93 is available from Danisco / DuPont under the trade name Puradax®. Other usable commercial products from Danisco / DuPont are Genencor detergent cellulase L and IndiAge® Neutra.

[0127] Other enzymes that can be used include, for example, lipases or cutinases, especially because of their triglyceride-cleaving activities, but also to produce suitable precursors. in situto produce peracids. These include, for example, those originally from Humicola lanuginosa (Thermomyces lanuginosus Lipases available from or further developed from these sources, in particular those with one or more of the following amino acid substitutions starting from the aforementioned lipase at positions D96L, T213R and / or N233R, especially T213R and N233R. Lipases are marketed, for example, by Novozymes under the trade names Lipolase®<, Lipolase®< Ultra, LipoPrime®<, Lipozyme®< and Lipex®<. Another advantageously usable lipase is available from Novozymes under the trade name Lipoclean®<. Furthermore, cutinases, originally derived from, for example, Fusarium solani pea and Humicola insolens have been isolated. Equally useful lipases are available from the company Amano under the names Lipase CE ®<, Lipase P ®<, Lipase B ®< or Lipase CES ®<, Lipase AKG ®< . Bacillus sp.Lipase®<, Lipase AP®<, Lipase M-AP®< and Lipase AML®< are available. Lipases and cutinases from the company Danisco / DuPont, for example, can be used, the precursor enzymes of which were originally derived from Pseudomonas mendocina and Fusarium solani have been isolated. Other important commercial products include the preparations M1 Lipase® and Lipomax® distributed by Danisco / DuPont, and the enzymes distributed by Meito Sangyo KK under the names Lipase MY-30®, Lipase OF® and Lipase PL®, as well as the product Lumafast® from Danisco / DuPont.

[0128] To enhance the bleaching effect, oxidoreductases, e.g., oxidases, oxygenases, catalases, peroxidases such as halo-, chloro-, bromo-, lignin-, glucose- or manganese-peroxidases, dioxygenases or laccases (phenol oxidases, polyphenol oxidases), can be used according to the invention. Advantageously, preferably organic, and especially aromatic, compounds that interact with the enzymes are additionally added to enhance the activity of the respective oxidoreductases (enhancers) or to ensure electron flow in cases of significantly different redox potentials between the oxidizing enzymes and the contaminants (mediators).

[0129] The enzymes to be used within the scope of the present invention can, for example, originally come from microorganisms, such as those of the genera Bacillus, Streptomyces, Humicola or Pseudomonas,originate and / or are produced by suitable microorganisms using known biotechnological methods, for example by transgenic expression hosts, e.g. of the genera Escherichia, Bacillus or by filamentous fungi. It is emphasized that these can also be technical enzyme preparations of the respective enzyme, meaning that accompanying substances may be present. Therefore, the enzymes can be formulated and used together with accompanying substances, for example from fermentation or with other stabilizers.

[0130] An agent according to the invention advantageously contains the protease in an amount of 2 µg to 20 mg, preferably 5 µg to 17.5 mg, particularly preferably 20 µg to 15 mg, and most preferably 50 µg to 10 mg per gram of the agent. In various embodiments, the concentration of the protease (active enzyme) described herein in the agent is >0 to 1 wt.%, preferably 0.001 to 0.1 wt.% based on the total weight of the agent.

[0131] Furthermore, the protease and / or other ingredients of the composition can be encapsulated in a substance that is impermeable to the enzyme at room temperature or in the absence of water, but which becomes permeable to the enzyme under the application conditions of the composition. Such an embodiment of the invention is thus characterized in that the protease is encapsulated in a substance that is impermeable to the protease at room temperature or in the absence of water. Furthermore, the detergent itself can also be packaged in a container, preferably an air-permeable container, from which it is released shortly before use or during the washing process.

[0132] The protease and / or any other enzymes of the same or different types contained in the detergent can be adsorbed onto carrier substances and / or embedded in coating materials to protect them against premature inactivation. In the washing solution, i.e., under application conditions, the enzyme is then released and can exert its catalytic effect. Preferred formulations contain 0.05 to 15 wt% and, in particular, up to 10 wt% of the active protein of the aforementioned protease.

[0133] A detergent according to the invention increasingly preferably contains the protease in an amount of 1 x 10 -8 to 5 wt.%, from 0.0001 to 1 wt.%, from 0.0005 to 0.5 wt.%, from 0.001 to 0.1 wt.%, each based on active protein.

[0134] In the preparations described herein, the enzymes to be used may also be formulated together with accompanying substances, such as those obtained from fermentation. In liquid formulations, the enzymes are preferably used as liquid enzyme formulation(s). The enzymes are generally not provided in the form of pure protein, but rather in the form of stabilized preparations that are suitable for storage and transport. These pre-formulated preparations include, for example, solid preparations obtained by granulation, extrusion, or lyophilization, or, particularly in the case of liquid or gel-like preparations, solutions of the enzymes, advantageously as concentrated as possible, with a low water content, and / or containing stabilizers or other excipients. Alternatively, the enzymes can be encapsulated for both solid and liquid dosage forms, e.g.,Enzymes are produced by spray drying or extrusion of the enzyme solution together with a preferably natural polymer, or in the form of capsules, e.g., capsules in which the enzymes are encapsulated as if in a solidified gel, or in core-shell type capsules in which an enzyme-containing core is coated with a protective layer impermeable to water, air, and / or chemicals. Additional active ingredients, e.g., stabilizers, emulsifiers, pigments, bleaching agents, or dyes, can be applied in superimposed layers. Such capsules are produced using methods known per se, e.g., by shake or roll granulation or in fluid-bed processes. Advantageously, such granules are low in dust and, due to the coating, have long-term storage stability, e.g., through the application of polymeric film formers. Furthermore, it is possible to combine two or more enzymes so that a single granule exhibits multiple enzyme activities.

[0135] The enzymes can also be incorporated into water-soluble films, such as those used in the formulation of unit-dose detergents. Such a film enables the release of the enzymes upon contact with water. As used herein, "water-soluble" refers to a film structure that is preferably completely water-soluble. Preferably, such a film consists of (fully or partially hydrolyzed) polyvinyl alcohol (PVA).

[0136] The embodiments of the present invention encompass all solid, powdered, granular, tablet-shaped, liquid, gel-shaped, or pasty dosage forms of the composition according to the invention, which may optionally consist of several phases and be in compressed or uncompressed form. The composition may be in the form of a free-flowing powder, in particular with a bulk density of 300 to 1,200 g / l, more specifically 500 to 900 g / l or 600 to 850 g / l. Solid dosage forms of the composition also include extrudates, granules, tablets, or pouches containing solid composition, which may be available in bulk containers or pre-packaged in individual portions. Alternatively, the composition may also be liquid, gel-shaped, or pasty, e.g., in the form of a non-aqueous composition or a non-aqueous paste, or in the form of an aqueous composition or a water-containing paste. Furthermore, the composition may be in the form of a single-component system.Such products consist of a single phase. Alternatively, a product can also consist of several phases (multi-component system). Such a product is therefore divided into several components, e.g., two liquid, two solid, or one liquid and one solid phase. The liquid formulations based on water and / or organic solvents can be thickened and presented in gel form.

[0137] A substance, e.g. a composition or an agent, is, according to the definition of the invention, solid if it is in the solid state of matter at 25°C and 1,013 mbar.

[0138] A substance, e.g., a composition or an agent, is liquid according to the definition of the invention if it exists in the liquid state at 25°C and 1,013 mbar. Liquid also includes gel-like states.

[0139] When the detergents according to the invention are in liquid form, they preferably contain more than 40 wt.%, preferably 50 to 90 wt.% and particularly preferably 60 to 80 wt.% water based on their total weight.

[0140] Another aspect of the invention is a method for cleaning textiles, wherein a textile detergent according to the invention, in particular a liquid textile detergent, is used in at least one process step. This includes both manual and machine methods, with machine methods being preferred due to their more precise controllability, for example, regarding the quantities used and contact times. Methods for cleaning textiles are generally characterized by the fact that, in one or more process steps, various cleaning-active substances are applied to the item to be cleaned and rinsed off after the contact time, or that the item to be cleaned is treated in some other way with a detergent or a solution thereof.

[0141] In various embodiments, the process is characterized by the fact that the protease is used at a temperature of 0 to 100°C, preferably 10 to 70°C, more preferably 30 to 50°C, and most preferably 20 to 40°C. In particular, in processes carried out at a temperature between 10 and 60°C, preferably between 15 and 50°C, and more preferably between 20 and 40°C, the advantages of using the textile detergents according to the invention compared to textile detergents with conventionally used proteases become apparent to the consumer.

[0142] All facts, objects, and embodiments described for textile detergents according to the invention are also applicable to this subject matter of the invention. Therefore, explicit reference is made here to the disclosure at the relevant point, with the note that this disclosure also applies to the aforementioned methods according to the invention.

[0143] Alternative embodiments of this invention also include processes for treating textile raw materials or for textile care, in which a protease according to the invention is activated in at least one process step. Processes for textile raw materials, fibers, or textiles with natural components are preferred, and especially for those containing wool or silk.

[0144] The proteases used in the compositions according to the invention can be advantageously used in textile detergents and textile washing processes, as described above. They can therefore be advantageously used to provide proteolytic activity in such compositions and processes.

[0145] Finally, the invention also encompasses the use of the proteases described herein in detergents, for example as described above, for the (improved) removal of protease-sensitive soils, for example from textiles. In preferred embodiments of this use, the protease is stored in the detergent for 3 or more days, 4 or more days, 7 or more days, 10 or more days, 12 or more days, 14 or more days, 21 or more days, or 28 or more days prior to a washing process.

[0146] Further inventions are therefore a use of an agent according to the invention for cleaning textiles, and / or a use of a protease comprising an amino acid sequence having at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its total length and, with reference to the numbering according to SEQ ID NO:1, (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, amino acid substitutions, in particular the amino acid substitutions 9T, 130D / V, 133A, 144K, 217M, 252T and 271, and (ii) at at least one, preferably at least two, of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, at least one further amino acid substitution, in particular selected from 6W / F, 89A / G, 131H / Y / F, 166M / L / I, 189T / L / I, 211N / Q and 224A / G, preferably selected from 6W, 89A, 131H, 166M, 189T, 211N and 224A, in a textile detergent, preferably a liquid textile detergent,for the removal of protease-sensitive soils from textiles, wherein the textile detergent has a pH value of approximately 9 to approximately 12, measured in a 1 wt% solution in deionized water at 20°C, in particular such that the protease is used in an amount of 2 µg to 20 mg, preferably 5 µg to 17.5 mg, particularly preferably 20 µg to 15 mg and most preferably 50 µg to 10 mg per gram of the agent.

[0147] Particularly preferred embodiments include, for example, hand washing, manual removal of stains from textiles, or use in conjunction with a machine process.

[0148] All circumstances, objects, and embodiments described for proteases and agents containing them according to the invention are also applicable to this subject matter of the invention. Therefore, explicit reference is made here to the disclosure at the relevant point, with the note that this disclosure also applies to the aforementioned use according to the invention. Examples Bestimmung der Lagerstabilität in liquidische Waschmittel Detergent matrix used

[0149] A commercially available liquid detergent, as specified in Table 1, was used. Table 1: Formula A Ingredients Formula A (in wt.%) LAS (anionic surfactant) 3% Alcohol Ethoxylate 7EO 2% Alcohol Ether Sulfate 3EO 8% Coconut fatty acids 0,5% NaOH 50% 0,4% Sodium carbonate 3% Distyrl biphenyl disulfonate 0,05% Fragrances 0,5% Demineralized water & misc.

[0150] The pH of Formula A was adjusted to pH 10 using citric acid, measured in a 1 wt. % solution in deionized water at 20°C.

[0151] The following proteases were used: V1: Protease according to SEQ ID NO:2 from WO 2013 / 060621 A1 V2: subtilisin 309 from Bacillus lentus E1: SEQ ID NO:1+P9T+S89A+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E storage

[0152] The proteases are located in bioreactor-generated supernatants in Bacillus licheniformis They are diluted to the same activity level. 90% detergent matrix was mixed with 10% appropriately diluted solution. Bacillus licheniformis The supernatant was added and thoroughly mixed. The enzyme activity in each batch was determined (starting value) before the sealed vessels were stored for two weeks at 30°C and 40°C, respectively. The extracted sample was dissolved in 0.1M Tris / HCl (pH 8.6) by stirring for 20 minutes at room temperature. The AAPF assay was then performed as described below. Protease activity assay (AAPF assay)

[0153] The activity of the protease is determined by the release of the chromophore para-nitroaniline from the substrate succinyl alanine-alanine-proline-phenylalanine-para-nitroanilide (AAPFpNA; Bachem L-1400). The release of pNA causes an increase in absorbance at 410 nm, the time course of which is a measure of the enzymatic activity.

[0154] The measurement was performed at a temperature of 25°C, pH 8.6, and a wavelength of 410 nm. The measurement time was 5 minutes with a measurement interval of 20 to 60 seconds. Measurement approach:

[0155] 10 µL AAPF solution (70 mg / mL) 1000 µL Tris / HCl (0.1 M; pH 8.6 with 0.1% Brij 35) 10 µL diluted protease solution Kinetics generated over 5 min at 25°C (410 nm) Results

[0156] The following shows the residual activity in % after storage in the aforementioned detergent matrix. The residual activity in % is relative to the respective enzyme activity before storage (starting value). Table 2: Storage in Formula B (pH 10) at 30°C or 40°C for two weeks Protease Residual activity (%) 30°C 40°C V1 18 3 V2 9 0 E1 33 6

[0157] The protease E1 according to the invention shows a significantly higher storage stability in a highly alkaline detergent formulation after storage for 2 weeks than the comparison proteases V1 and V2.

Claims

1. Textile detergent, in particular liquid textile detergent, comprising a) at least one protease, wherein the protease exhibits proteolytic activity and comprises an amino acid sequence that exhibits at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its entire length and, in each case, relative to the numbering according to SEQ ID NO:1 (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252, and 271, amino acid substitutions, in particular the amino acid substitutions 9T, 130D / V, 133A, 144K, 217M, 252T and 271E, and (ii) at at least one, preferably at least two, of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, at least one further amino acid substitution, in particular selected from 6W / F, 89A / G, 131H / Y / F, 166M / L / I, 189T / L / I, 211N / Q and 224A / G, preferably selected from 6W, 89A, 131H, 166M, 189T, 211N, and 224A, and b) at least one detergent ingredient, preferably in an amount of 0.01 to 99.9 % by weight, wherein the textile detergent has a pH value about 9 to about 12, measured in a 1 % by weight solution in deionized water at 20°C.

2. Detergent according to claim 1, wherein the protease comprises an amino acid sequence that is at least 70% and increasingly preferably at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, and 96.5%, and in each case, based on the numbering according to SEQ ID NO:1 (i), at the positions corresponding to positions 9, 130, 133, 144, 217, 252 and 271, the amino acid substitutions 9T, 130D / V, 133A, 144K, 217M, 252T and 271E, and (ii) at at least two of the positions corresponding to positions 6, 89, 131, 166, 189, 211, or 224, at least two further amino acid substitutions selected from the group consisting of 6W / F, 89A / G, 131H / Y / F, 166M / L / I, 189T / L / I, 211N / Q, and 224A / G, preferably 6W, 89A, 131H, 166M, 189T, 211N and 224A, preferably 6W, 89A, 131H, 166M, 189T, 211N and 224A.

3. Detergent according to claim 1 or 2, wherein the protease has one of the following amino acid substitution variants, each based on the numbering according to SEQ ID NO:1: (i) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+N252T+Q271E; (ii) P9T+N130D+T133A+N144K+G166M+S189T+Y217M+S224A+N252T+Q271E; (iii) P9T+S89A+N130D+G131H+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (iv) Y6W+P9T+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E; (v) P9T+N130D+T133A+N144K+G166M+S211N+Y217M+N252T+Q271E; (vi) P9T+S89A+N130D+T133A+N144K+S189T+Y217M+S224A+N252T+Q271E.

4. Detergent according to any one of claims 1 to 3, wherein the protease is present in an amount of 1 x 10-8 to 5 % by weight and increasingly preferably from 0.0001 to 1 % by weight, from 0.0005 to 0.5 % by weight, from 0.001 to 0.1 % by weight, based on the total weight of the detergent contained therein.

5. Detergent according to any one of claims 1 to 4, wherein the agent comprises at least one further enzyme selected from the group consisting of amylases, cellulases, hemicellulases, mannanases, tannases, xylanases, xanthanases, xyloglucanases, β-glucosidases, pectinases, carrageenases, perhydrolases, oxidases, oxidoreductases, lipase, and combinations thereof, preferably at least one amylase.

6. Detergent according to claim 5, wherein the at least one further enzyme is an amylase and wherein the amylase is selected from: a) α -amylase comprising an amino acid sequence that is at least 80% identical to the amino acid sequence specified in SEQ ID NO:2 over its entire length and optionally has at least one amino acid substitution at one of positions 172, 202, 208, 255, and 261 in the numbering according to SEQ ID NO:2, preferably selected from the group consisting of M202L, M202V, M202S, M202T, M202I, M202Q, M202W, S255N, R172Q, and combinations thereof; and / or b) α-amylase comprising an amino acid sequence that is at least 60% identical to the amino acid sequence specified in SEQ ID NO:3 over its entire length and optionally has at least one amino acid substitution at one of positions 9, 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 195, 202, 203, 214, 231, 256, 257, 258, 269, 270, 272, 283, 295, 296, 298, 299, 303, 304, 305, 311, 314, 315, 318, 319, 320, 323, 339, 345, 361, 378, 383, 419, 421, 437, 441, 444, 445, 446, 447, 450, 458, 461, 471, 482 and 484 and / or a deletion at one of positions 183 and 184 in the counting according to SEQ ID NO:3, preferably at least one amino acid substitution at one of positions 9, 26, 149, 182, 186, 202, 257, 295, 299, 323, 339, and 345, and / or particularly preferably at least one amino acid substitution or deletion selected from the group consisting of R118K, D183*, G184*, N195F, R320K, R458K, and combinations thereof; and / or c) α-amylase comprising an amino acid sequence that is at least 90% identical to the amino acid sequence specified in SEQ ID NO:4 over its entire length and optionally has at least one substitution and / or deletion at one of positions 93, 116, 118, 129, 133, 134, 140, 142, 146, 147, 149, 151, 152, 169, 174, 183, 184, 186, 189, 193, 195, 197, 198, 200, 203, 206, 210, 212, 213, 235, 243, 244, 260, 262, 284, 303, 304, 320, 338, 347, 359, 418, 431, 434, 439, 447, 458, 469, 476, and 477 in the numbering according to SEQ ID NO:4, preferably amino acid deletions at positions 183 and 184.

7. Detergent according to any one of claims 1 to 6, wherein the detergent ingredient is selected from the group consisting of surfactants, builders, complexing agents, polymers, glass corrosion inhibitors, corrosion inhibitors, bleaching agents such as peroxygen compounds, bleach activators or bleach catalysts, water-miscible organic solvents, enzyme stabilizers, sequestrants, electrolytes, pH regulators, and / or other auxiliaries such as optical brighteners, gray inhibitors, color transfer inhibitors, foam regulators, and dyes and fragrances, as well as combinations thereof.

8. Detergent according to any of claims 1 to 7, comprising a) 5 to 20 % by weight, in particular 9 to 17 % by weight, of anionic surfactants and / or b) 1 to 20 % by weight, in particular 5 to 9 % by weight, of nonionic surfactants and / or c) 0.5 to 10 % by weight, in particular 1 to 4 % by weight, water-soluble organic builder substances, in particular citric acid or citrate, and / or d) 0.01 to 5 % by weight, in particular 0.5 to 2 % by weight, of complexing agents, in particular HEDP, and / or e) 0.01 to 5 % by weight, in particular 0.2 to 1 % by weight, of a gray inhibitor and / or f) 0.01 to 5 % by weight of a foam inhibitor.

9. Detergent according to any one of claims 1 to 7, comprising a) 0 to 10 % by weight, in particular 1 to 3 % by weight, of anionic surfactants and / or b) 5 to 20 % by weight, in particular 10 to 18 % by weight, of nonionic surfactants and / or c) 1 to 10 % by weight, in particular 1.5 to 3 % by weight, of water-soluble or water-dispersible inorganic builder material and / or d) 0 to 1 % by weight, in particular 0.01 to 0.5 % by weight, of fragrances and / or e) 0.01 to 5 % by weight, in particular 0.05 to 0.5 % by weight, optical brighteners.

10. Detergent according to one of claims 1 to 9, wherein it has a pH value in a range of about 9 to about 12, in particular from about 9.5 to about 11.5, more preferably from about 10 to about 11, particularly preferably from about pH 10, in a 1 % by weight solution in deionized water at 20°C.

11. Method for cleaning textiles, characterized in that an agent according to one of claims 1 to 10 is used in at least one step of the method.

12. Use of a detergent according to one of claims 1 to 10 for removing protease-sensitive soiling from textiles.

13. Use of a protease comprising an amino acid sequence which has at least 70% sequence identity with the amino acid sequence specified in SEQ ID NO:1 over its entire length and, in each case, based on the numbering according to SEQ ID NO:1 (i) at the positions corresponding to positions 9, 130, 133, 144, 217, 252, and 271, amino acid substitutions, in particular the amino acid substitutions 9T, 130D / V, 133A, 144K, 217M, 252T and 271E, and (ii) at at least one, preferably at least two, of the positions corresponding to positions 6, 89, 131, 166, 189, 211 or 224, at least one further amino acid substitution, in particular selected from 6W / F, 89A / G, 131H / Y / F, 166M / L / I, 189T / L / I, 211N / Q and 224A / G, preferably selected from 6W, 89A, 131H, 166M, 189T, 211N and 224A, in a textile detergent, preferably a liquid textile detergent, for removing protease-sensitive soiling from textiles, wherein the textile detergent has a pH value of about 9 to about 12, measured in a 1 % by weight solution in deionized water at 20°C.