WO2025113890A1 - Washing and cleaning composition with protease - Google Patents

Abstract

The disclosure is in the field of detergents and the invention particularly relates to washing and cleaning composition, comprising at least one protease, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1, (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161, 163, 171, 181, 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitutions) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q; less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, and at least one surfactant.

Description

WASHING AND CLEANING COMPOSITION WITH PROTEASE

DESCRIPTION

The disclosure is in the field of detergents, in particular detergents comprising at least one protease. The disclosure relates to a textile detergent, in particular a liquid textile detergent, comprising at least one protease of which the amino acid sequence has been modified in particular with regard to use in textile detergents, and at least one surfactant, the textile detergent comprising less than 0.2 wt.% boric acid, preferably being free of boric acid. Also part of the disclosure are the corresponding washing methods, the use of the compositions described herein and the use of proteases in textile detergents comprising less than 0.2 wt.% boric acid, preferably being free of boric acid, and the use of proteases in textile detergents comprising less than 0.2 wt.% boric acid, preferably being free of boric acid.

The most important criterion when cleaning textiles and/or hard surfaces is the cleaning performance on a wide variety of stains. Even though the cleaning performance of the washing and cleaning compositions used today is generally high, the general trend towards the increased use of low-temperature programs and the trend towards compacting formulations (especially for preportioned forms of use) has led to the problem that many of the usual washing and cleaning composition formulations have insufficient cleaning performance, especially on stubborn stains soiling. Such inadequate cleaning performance leads to consumer dissatisfaction and to such stains being pre-treated by the consumer, which in turn increases water and energy consumption. Hence, constantly changing requirements are being placed on the packaging and supply forms of washing and cleaning compositions. The compaction of modern washing and cleaning compositions requires the ongoing optimization of existing formulations and the development of new, more efficient formulations. One aim of these developments, which generally make use of known active washing and cleaning ingredients, is to increase the washing or cleaning performance of the formulations while maintaining or reducing the dosage.

The use of enzymes in detergents has been established in the prior art for decades. They are used to expand the performance range of the compositions in question according to their special activities. These include in particular hydrolytic enzymes such as proteases, amylases, lipases and cellulases. The first three mentioned hydrolyze proteins, starch and fats and thus contribute directly to the removal of dirt. Cellulases are used in particular due to their effect on fabric. Another group of detergent enzymes are oxidative enzymes, in particular oxidases, which, optionally in conjunction with other components, are preferably used to bleach stains or to produce the bleaching agents in situ. In addition to these enzymes, which are subject to continuous optimization, other enzymes such as pectinases, p-glucanases, mannanases or other hemicellulases (glycosidases) are constantly being made available for use in detergents in particular in order to be able to optimally tackle specific stains, to hydrolyze specific vegetable polymers in particular. Proteases are among the technically most important enzymes, and they are the longest- established enzymes contained practically all modern effective detergents. Of these, in turn, proteases of the subtilisin type (subtilases, subtilopeptidases, EC 3.4.21.62), which are serine proteases due to the catalytically active amino acids, are particularly important. They act as non-specific endopeptidases and hydrolyze any acid amide bonds that are inside peptides or proteins. Their optimum pH is usually in the alkaline range at roughly pH 9. An overview of this family is given, e.g., in the article "Subtilases: Subtilisin-like Proteases" by R. Siezen, pages 75-95 in "Subtilisin enzymes", published by R. Bott and C. Betzel, New York, 1996. Subtilases are formed naturally from microorganisms. In particular, the subtilisins formed and secreted by Bacillus species are the most significant group of subtilases. Examples of the subtilisin-type proteases preferably used in washing and cleaning compositions are the subtilisins BPN' from Bacillus amyloliquefaciens and Carlsberg from Bacillus licheniformis, the protease PB92, the subtilisins 147 and 309, the protease from Bacillus lentus, in particular from Bacillus lentus DSM 5483, 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 narrower sense, as well as variants of the aforementioned proteases which have a different amino acid sequence to the parent protease. Proteases are modified in a targeted or random manner using processes known from the prior art and thus optimized for use in washing and cleaning compositions. These include point, deletion or insertion mutagenesis or fusion with other proteins or protein parts. Correspondingly optimized variants are known for most proteases known from the prior art. EP 2016175, e.g., discloses a protease from Bacillus pumilus intended for washing and cleaning compositions. WO 2013/060621 and EP 3660151 describe protease variants of the alkaline protease from Bacillus lentus that are optimized for washing and cleaning compositions. International patent applications WO 95/23221 A1 , WO 92/21760 A1 and WO 2013/060621 A1 disclose variants of the protease from Bacillus lentus DSM 5483 which are suitable for use in washing or cleaning compositions. Furthermore, the international patent applications WO 2011/032988 A1 and WO 2016/096714 A1 and the European patent application EP 3044302 A1 disclose washing and cleaning compositions which contain variants of the protease from Bacillus lentus DSM 5483. The protease variants disclosed in these documents can, in addition to other positions, be modified at positions 3, 4, 99 and/or 199 in the numbering of the protease from Bacillus lentus DSM 5483 and, e.g., have the amino acids 3T, 4I, 99E or 1991 at said positions. However, it is not clear from these documents that certain Bacillus lentus protease variants are suitable for use in washing or cleaning compositions comprising only low amounts of boric acid, preferably being free of boric acid.

In washing and cleaning compositions, proteases are used to break down protein-containing stains on the items to be cleaned. However, they also hydrolyze themselves (autoproteolysis) and all other proteins contained in the respective compositions, i.e., in particular other enzymes contained in the washing and cleaning compositions. This occurs particularly during the cleaning process, i.e., in the aqueous washing or cleaning liquor, when comparatively favorable reaction conditions are present. To a lesser extent, however, this also occurs during storage of the respective compositions, which is why long storage is always accompanied by a certain loss of protease activity and the activity of other enzymes. Due to the loss of enzymatic activity, they no longer exhibit optimum cleaning performance. This is particularly problematic in gel or liquid formulations and especially in formulations containing water because the water contained in these formulations provides both the reaction medium and the hydrolysis reagent.

In general, only selected proteases are suitable for use in liquid preparations containing surfactants. Many proteases do not show sufficient catalytic performance in such preparations or they are not sufficiently stable. For the application of proteases in detergents, a high catalytic activity and stability under conditions as they occur during a washing process is therefore particularly desirable.

One aim in the development of washing and cleaning agent formulations is therefore not only to stabilize the contained enzymes, particularly during storage, but also to protect them from denaturation and/or cleavage or degradation and/or decomposition due to physical influences or oxidation, etc., particularly during storage and/or use of the washing or cleaning composition. One focus of these developments is the protection of the proteins and/or enzymes contained against (auto-)proteolytic cleavage. This can be achieved by creating physical barriers, e.g., by encapsulating the enzymes in special enzyme granules or by packaging the compositions in two- or multi-chamber systems. The other frequently used method is to add chemical compounds that inhibit the proteases and thus act as stabilizers or inhibitors for proteases and other proteins and enzymes contained in the product. However, these must be reversible protease inhibitors, as the protease activity should only be temporarily inhibited, particularly during storage, but no longer during the cleaning process.

Various reversible protease inhibitors are described in the prior art, e.g., boric acids, boronic acids or derivatives, salts or esters thereof, in particular 4-formylphenylboronic acid (4-FPBA), as well as peptide aldehydes, i.e., oligopeptides with a reduced C-terminus, in particular those consisting of 2 to 50 monomers.

Due to sustainability efforts and concerns about the environmental compatibility of boric acid and/or other boron-containing substances in washing and cleaning compositions, more and more washing and cleaning compositions are being developed that contain less to no boron-containing compounds, especially boric acid.

However, there is still a need to improve the cleaning performance of enzyme-containing washing and cleaning compositions and to better stabilize the enzymes contained in washing and cleaning compositions. In particular, there is an increasing requirement to avoid boric acid-containing compounds, i.e., in particular boron-containing protease inhibitors, in washing and cleaning compositions.

Surprisingly, it was found that a protease having proteolytic activity and comprising an amino acid sequence which 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%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, is more stable in washing and cleaning compositions with low boric acid contents, in particular boric acid free washing and cleaning compositions. More surprisingly, such proteases do not seem to need any of the classical reversible protease inhibitors like boric acid or 4-FPBA or the like.

In a first aspect, the invention therefore relates to a washing and cleaning composition, preferably liquid washing and cleaning composition, more preferably liquid laundry detergent, comprising, each based on the total weight of the washing and cleaning composition, at least one protease, in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein, wherein the protease has proteolytic activity and comprises an amino acid sequence which 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%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, and optionally at least one further enzyme, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases, pectate lyases and mixtures thereof, each in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein. In a further aspect, the invention relates to a washing and cleaning composition, preferably liquid washing and cleaning composition, more preferably liquid laundry detergent, comprising, each based on the total weight of the washing and cleaning composition, at least one protease, in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71 %, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 and 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K and L256E; less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, and optionally at least one further enzyme, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases, pectate lyases and mixtures thereof, each in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein.

In a further aspect, the invention relates to a washing and cleaning composition, preferably liquid washing and cleaning composition, more preferably liquid laundry detergent, comprising, each based on the total weight of the washing and cleaning composition, at least one protease, in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations: S3T-V4I-R99E-V199I-Q200L-Y203W; S3T-V4I- R99E-V199I-N212S; S3T-V4I-R99E-V199I-N74D; S3T-V4I-R99E-V199I-S154D-L256E; S3T-V4I- R99E-V199I-Q200L-Y203W-S154D-L256E; S3T-V4I-R99E-V199I-N74D-Q200L-Y203W; S3T-V4I- R99E-V199I-N74D-S154D-Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-N74D-N212S; S3T-V4I- R99E-V199I-N74D-S154D-Y203W-L256E; S3T-V4I-R99E-V199I-N74D-Y203W; S3T-V4I-R99E- V199I-N74D-S154D-Q200L-L256E; S3T-V4I-R99E-V199I-N74D-Q200L; S3T-V4I-R99E-V199I-

S154D-Q200L-Y203W; S3T-V4I-R99E-V199I-Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-A136Q-

R143W-Y161 T-Q200L; S3T-V4I-R99E-V199I-N74D-R143Y-A209W-N212S-L256E; S3T-V4I-R99E- V199I-A136Q-S154D-V171 L-Q200L; S3T-V4I-R99E-V199I-Q200L-Y203W-A209K-S154D-L256E;

S3T-V4I-R99E-V199I-S154D-S160G-Q185R-Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-S1540-

Al 81 D-F183R-Q200L-Y203W-L256E; S3T-V4I-V199I-Q200L-Y203W; S3T-V4I-V199I-Q200L-Y203W- S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y- A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200L- Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209K-S154D-L256E; S3T-V4I-V1991- Q200I-Y203Y-A209K-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E; less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, and optionally at least one further enzyme, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases, pectate lyases and mixtures thereof, each in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein.

In preferred embodiments, the washing and cleaning composition described herein is free of boric acid.

In preferred embodiments, the washing and cleaning composition described herein is essentially free, preferably free, of any protease stabilizers selected from the group consisting of boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4-formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, the washing and cleaning composition described herein comprises at least one protease according to any one of claims 1 to 3 in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and

LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

Further aspects of the invention relate to

• a method for cleaning textiles and/or hard surfaces, characterized in that a washing or cleaning composition described herein is used in at least one method step, the method preferably being carried out in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C;

• use of a washing or cleaning composition described herein for cleaning textiles and/or hard surfaces, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C;

• use of a washing or cleaning composition described herein for improving the stability, in particular the storage stability, of enzymes, preferably proteases, in a washing and cleaning composition, preferably in a washing or cleaning composition described herein;

• use of a washing or cleaning composition described herein for preventing and/or reducing the proteolysis of further enzymes contained in the washing and cleaning composition by a protease contained in the washing and cleaning composition, wherein the washing and cleaning composition is preferably a washing and cleaning composition described herein;

• use of a washing or cleaning composition described herein for preventing and/or reducing the autoproteolysis of a protease contained in the washing and cleaning composition, wherein the washing and cleaning composition is preferably a washing and cleaning composition described herein.

Preferably, the washing and cleaning composition according to the invention is a liquid textile washing composition. Further preferably, the textile washing composition according to the invention in 1 wt.% solution in deionized water at 20°C has a pH value in a range from about 6 to about 11 , preferably from about 6.5 to about 10.5, more preferably from about 7 to about 10, most preferably from about 8 to about 9.

Textile washing compositions according to the invention also particularly show performance advantages over other textile washing compositions if the textile washing compositions contain at least one additional enzyme of the same or a different kind, e.g., amylase, cellulase, lipase, mannanase or pectinase, the list of further enzymes not being exhaustive. It is therefore preferred that the textile washing compositions according to the invention contain at least one additional enzyme of the same type (i.e., a further protease) or of a different type, preferably selected from amylase, cellulase, mannanase and lipase.

These and other aspects, features and advantages of the invention will become apparent to the person skilled in the art from a study of the following detailed description and claims. Any feature from one aspect of the invention can be used in any other aspect of the invention. Furthermore, it is understood that the examples contained herein are intended to describe and illustrate the invention, but are not limiting thereto and, in particular, the invention is not limited to these examples.

All percentages are percent by weight (wt.%) unless otherwise indicated.

Numerical ranges given in the format "from x to y" include the stated values. Where multiple preferred numerical ranges are given in this format, it is understood that all ranges resulting from the combination of the various endpoints are also included.

"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. The term "washing or cleaning agent" or "washing and cleaning composition" as used herein is synonymous with the term "agent" or "detergent" and means a composition for cleaning textiles and/or hard surfaces, in particular dishes, as explained in the description.

"About" or "approximately" as used herein with respect to a numerical value refers to the corresponding numerical value ±10%, preferably ±5%.

"Liquid" as used herein includes liquids and gels as well as pasty compositions. It is preferred that the liquid compositions are flowable and pourable at room temperature, but it is also possible that they have a yield point.

A substance, e.g., a composition or an agent, is solid according to the definition of the invention if it is in the solid state at 25°C and 1013 mbar.

A substance, e.g., a composition or an agent, is liquid according to the definition of the invention if it is in the liquid state at 25°C and 1013 mbar. In this context, liquid also includes gel-like.

"Variant" as used herein refers to natural or artificially produced variations of a native enzyme or peptide having an amino acid sequence that is modified from the reference form.

The term "textile" as used herein means any textile material, including yarns, yarn intermediates, fibers, nonwovens, natural materials, synthetic materials, and any other textile materials, fabrics made from these materials, and products made from fabrics (e.g., garments and other articles). The textile or fabric can be in the form of knitted fabrics, woven fabrics, denims, nonwovens, felts, yarns and terry cloth. The textile may be cellulose-based, such as natural cellulosic fibers such as cotton, flax/linen, jute, ramie, sisal or coir, or man-made cellulosic fibers (e.g., from wood pulp) such as viscose/rayon, cellulose acetate fibers (tricell), lyocell or blends thereof. The textile or fabric may also consist of non- cellulosic fibers, e.g., natural polyamides such as wool, camel, cashmere, mohair, rabbit and silk or synthetic polymers such as nylon, aramid, polyester, acrylic, polypropylene and spandex/elastane or blends thereof as well as blends of cellulosic fibers and non-cellulosic fibers. Examples of blends are blends of cotton and/or rayon/viscose with one or more accompanying materials such as wool, synthetic fibers (e.g., polyamide fibers, acrylic fibers, polyester fibers, polyvinyl chloride fibers, polyurethane fibers, polyurea fibers, aramid fibers) and/or cellulosic fibers (e.g., rayon/viscose, ramie, flax/linen, jute, cellulose acetate fibers, lyocell). The fabric may be conventional washable laundry, e.g., soiled household linen. When the term "fabric" or "garment" is used, it should also include the broader term "textiles".

An "improvement of the stability of an enzyme" within the meaning of the invention is present if the protease defined herein exhibits a higher enzymatic activity of the protease and/or optionally further enzymes contained in the washing or cleaning composition after storage in a washing or cleaning composition according to the invention compared to a reference preparation comprising a washing or cleaning composition not according to the invention and/or comprising a protease not according to the invention. After storage, the washing or cleaning composition according to the invention therefore has a higher residual activity of the contained protease and/or possibly other contained enzymes compared to the reference, wherein the washing or cleaning composition according to the invention and the reference have the same initial enzymatic activity at the start of storage, both samples are treated in the same way, in particular with regard to the conditions of storage and the determination of the enzyme activity. Increasingly preferably, storage is carried out for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or 8 weeks. Further preferably the storage is carried out at a temperature of 20°C, 25°C, 30°C or 40°C.

The present invention is based on the surprising finding of the inventors that proteases described herein are more stable in washing and cleaning compositions with low boric acid contents, in particular boric acid free washing and cleaning compositions. More surprisingly, such proteases do not seem to need any of the classical reversible protease inhibitors like boric acid or 4-FPBA or peptides or the like. This is insofar surprising since the addition of stabilizers or reversible protease inhibitors (e.g., boric acid, 4-FPBA, glycerol, 1 ,2-propanediol, peptide aldehydes, or the like) in washing or cleaning compositions is usually necessary to ensure that proteases in washing or cleaning compositions are stable and to prevent proteases from degrading themselves or other enzymes contained in the washing or cleaning composition.

In preferred embodiments, a washing and cleaning composition according to the invention comprises, each based on the total weight of the washing and cleaning composition, at least one protease, in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein, wherein the protease has proteolytic activity and comprises an amino acid sequence which 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%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, and optionally at least one further enzyme, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases, pectate lyases and mixtures thereof, each in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein.

In preferred embodiments, a composition according to the invention is characterized in that it comprises a protease as described herein. Preferably, the composition is a washing and cleaning composition, in particular a washing and cleaning composition containing less than 0.2 wt.%, preferably a boric acid-free washing and cleaning composition, particularly preferably a boric acid-free textile washing composition. More preferably, the composition is a boron-free washing and cleaning composition, in particular a boron-free textile detergent.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E. In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E. In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199l, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199l, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E.

In particularly preferred embodiments, the washing and cleaning compositions according to the invention comprise a protease, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which comprises, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations: S3T-V4I-R99E-V199I-Q200L-Y203W; S3T-V4I-R99E-V199I-N212S; S3T-V4I-R99E- V199I-N74D; S3T-V4I-R99E-V199I-S154D-L256E; S3T-V4I-R99E-V199I-Q200L-Y203W-S154D- L256E; S3T-V4I-R99E-V199I-N74D-Q200L-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L- Y203W-L256E; S3T-V4I-R99E-V199I-N74D-N212S; S3T-V4I-R99E-V199I-N74D-S154D-Y203W- L256E; S3T-V4I-R99E-V199I-N74D-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-L256E; S3T- V4I-R99E-V199I-N74D-Q200L; S3T-V4I-R99E-V199I-S154D-Q200L-Y203W; S3T-V4I-R99E-V199I- Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-A136Q-R143W-Y161 T-Q200L; S3T-V4I-R99E-V1991- N74D-R143Y-A209W-N212S-L256E; S3T-V4I-R99E-V199I-A136Q-S154D-V171 L-Q200L; S3T-V4I- R99E-V199I-Q200L-Y203W-A209K-S154D-L256E; S3T-V4I-R99E-V199I-S154D-S160G-Q185R- Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-S154D-A181 D-F183R-Q200L-Y203W-L256E; S3T-V4I- V199I-Q200L-Y203W; S3T-V4I-V199I-Q200L-Y203W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W- A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209W-S154D-L256E; S3T-V4I-V199I-Q200I- Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209W-S154D-L256E; S3T-V4I- V199I-Q200I-Y203W-A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209K-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E.

In particularly preferred embodiments, the washing and cleaning compositions according to the invention comprise a protease, wherein the protease comprises, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations: S3T-V4I-R99E- V199I-Q200L-Y203W; S3T-V4I-R99E-V199I-N212S; S3T-V4I-R99E-V199I-N74D; S3T-V4I-R99E- V199I-S154D-L256E; S3T-V4I-R99E-V199I-Q200L-Y203W-S154D-L256E; S3T-V4I-R99E-V199I- N74D-Q200L-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-N74D-N212S; S3T-V4I-R99E-V199I-N74D-S154D-Y203W-L256E; S3T-V4I-R99E-V199I-N74D- Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-L256E; S3T-V4I-R99E-V199I-N74D-Q200L; S3T- V4I-R99E-V199I-S154D-Q200L-Y203W; S3T-V4I-R99E-V199I-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-A136Q-R143W-Y161 T-Q200L; S3T-V4I-R99E-V199I-N74D-R143Y-A209W-N212S-L256E; S3T-V4I-R99E-V199I-A136Q-S154D-V171 L-Q200L; S3T-V4I-R99E-V199I-Q200L-Y203W-A209K- S154D-L256E; S3T-V4I-R99E-V199I-S154D-S160G-Q185R-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-S154D-A181 D-F183R-Q200L-Y203W-L256E; S3T-V4I-V199I-Q200L-Y203W; S3T-V4I-V199I- Q200L-Y203W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E; S3T-V4I-V1991- Q200I-Y203Y-A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209W-S154D-L256E; S3T-V4I- V199I-Q200L-Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209K-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D- L256E, wherein the protease comprises no further changes in addition to the amino acid substitutions mentioned.

In preferred embodiments, the amino acid substitutions and/or amino acid substitution combinations described herein lead to an improved storage stability of this modified protease in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, preferably boric acid-free textile washing compositions. In particularly preferred embodiments, the amino acid substitutions and/or amino acid substitution combinations described herein lead to an improved storage stability of this modified protease in boron-free washing and cleaning compositions, in particular boron-free textile detergents.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two of the positions corresponding to positions 3, 4, 99 or 199, at least two amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least three of the positions corresponding to positions 3, 4, 99 or 199, at least three amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one amino acid substitution selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least two of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least two amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least three of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least three amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least four of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least four amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In preferred embodiments the protease used in washing and cleaning compositions according to the invention is a protease that has proteolytic activity and comprises an amino acid sequence which 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%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 or 199, the amino acid substitutions S3T, V4I, R99E and V199l, and (ii) at at least five of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least five amino acid substitutions selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In particularly preferred embodiments, the washing and cleaning compositions according to the invention comprise a protease, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which comprises, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations: S3T-V4I-R99E-V199I-Q200L-Y203W; S3T-V4I-R99E-V199I-N212S; S3T-V4I-R99E- V199I-N74D; S3T-V4I-R99E-V199I-S154D-L256E; S3T-V4I-R99E-V199I-Q200L-Y203W-S154D- L256E; S3T-V4I-R99E-V199I-N74D-Q200L-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L- Y203W-L256E; S3T-V4I-R99E-V199I-N74D-N212S; S3T-V4I-R99E-V199I-N74D-S154D-Y203W- L256E; S3T-V4I-R99E-V199I-N74D-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-L256E; S3T- V4I-R99E-V199I-N74D-Q200L; S3T-V4I-R99E-V199I-S154D-Q200L-Y203W; S3T-V4I-R99E-V199I- Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-A136Q-R143W-Y161 T-Q200L; S3T-V4I-R99E-V1991- N74D-R143Y-A209W-N212S-L256E; S3T-V4I-R99E-V199I-A136Q-S154D-V171 L-Q200L; S3T-V4I- R99E-V199I-Q200L-Y203W-A209K-S154D-L256E; S3T-V4I-R99E-V199I-S154D-S160G-Q185R- Q200L-Y203W-L256E; S3T-V4I-R99E-V199I-S154D-A181 D-F183R-Q200L-Y203W-L256E; S3T-V4I- V199I-Q200L-Y203W; S3T-V4I-V199I-Q200L-Y203W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W- A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209W-S154D-L256E; S3T-V4I-V199I-Q200I- Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209W-S154D-L256E; S3T-V4I- V199I-Q200I-Y203W-A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209K-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions, wherein the stability is determined as described in example 2.

In particularly preferred embodiments, the washing and cleaning compositions according to the invention comprise a protease, wherein the protease comprises, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations: S3T-V4I-R99E- V199I-Q200L-Y203W; S3T-V4I-R99E-V199I-N212S; S3T-V4I-R99E-V199I-N74D; S3T-V4I-R99E- V199I-S154D-L256E; S3T-V4I-R99E-V199I-Q200L-Y203W-S154D-L256E; S3T-V4I-R99E-V199I- N74D-Q200L-Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-N74D-N212S; S3T-V4I-R99E-V199I-N74D-S154D-Y203W-L256E; S3T-V4I-R99E-V199I-N74D- Y203W; S3T-V4I-R99E-V199I-N74D-S154D-Q200L-L256E; S3T-V4I-R99E-V199I-N74D-Q200L; S3T- V4I-R99E-V199I-S154D-Q200L-Y203W; S3T-V4I-R99E-V199I-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-A136Q-R143W-Y161 T-Q200L; S3T-V4I-R99E-V199I-N74D-R143Y-A209W-N212S-L256E; S3T-V4I-R99E-V199I-A136Q-S154D-V171 L-Q200L; S3T-V4I-R99E-V199I-Q200L-Y203W-A209K- S154D-L256E; S3T-V4I-R99E-V199I-S154D-S160G-Q185R-Q200L-Y203W-L256E; S3T-V4I-R99E- V199I-S154D-A181 D-F183R-Q200L-Y203W-L256E; S3T-V4I-V199I-Q200L-Y203W; S3T-V4I-V199I- Q200L-Y203W-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E; S3T-V4I-V1991- Q200I-Y203Y-A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209W-S154D-L256E; S3T-V4I- V199I-Q200L-Y203W-A209W-S154D-L256E; S3T-V4I-V199I-Q200I-Y203W-A209K-S154D-L256E; S3T-V4I-V199I-Q200I-Y203Y-A209K-S154D-L256E; S3T-V4I-V199I-Q200L-Y203W-A209K-S154D- L256E, wherein the protease comprises no further changes in addition to the amino acid substitutions mentioned, wherein the protease exhibits improved storage stability in washing or cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid- free textile washing compositions, wherein the stability is determined as described in example 2.

Proteases used in compositions according to the invention have an increased catalytic activity in washing and cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid-free textile washing compositions. In various embodiments, the proteases used in compositions according to the invention can have a proteolytic activity which is higher than that of the wild type (SEQ ID NO:1) and/or an already performance-improved starting variant of the protease is at least 101 %, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130% or more. Such performance-enhanced proteases enable improved cleaning results on protease-sensitive stains. Such performance-improved cleaning results on proteasesensitive stains can be achieved in various temperature ranges, e.g., in a range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

The proteases used according to the invention have a high stability in washing and cleaning compositions, in particular washing and cleaning compositions containing less than 0.2 wt.% boric acid, preferably boric acid-free washing and cleaning compositions, particularly preferably boric acid- free textile washing compositions. In preferred embodiments, proteases used in compositions according to the invention also have a high stability, e.g. towards surfactants and/or chelators, and/or towards temperature influences, in particular towards high temperatures, e.g. between about 50°C and about 65°C, in particular about 60°C, and/or towards acidic or alkaline conditions and/or towards pH changes and/or towards denaturing or oxidizing agents and/or towards (auto-)proteolytic degradation and/or towards a change in the redox ratios.

The proteases used according to the invention have enzymatic activity, i.e., they are capable of hydrolyzing peptides and proteins, in particular in washing and cleaning compositions. A protease used in compositions according to the invention is therefore an enzyme which catalyzes the hydrolysis of amide/peptide bonds in protein/peptide substrates and is thus capable of cleaving proteins or peptides. Furthermore, a protease used in compositions according to the invention is preferably a mature protease, i.e., the catalytically active molecule without signal and/or propeptide(s). Unless otherwise indicated, the sequences given herein refer to mature (processed) enzymes.

In various embodiments of the invention, the protease is a freely present enzyme. This means that the protease can act directly with all components of a composition and, if the composition is a liquid composition, that the protease is in direct contact with the solvent of the composition (e.g., water). In other embodiments, a composition may comprise proteases that form an interaction complex with other molecules or that comprise an "envelope". Here, a single or multiple protease molecules may be separated from the other components of the composition by a surrounding structure. Such a separating structure may be formed by, but is not limited to, vesicles, such as a micelle or a liposome. However, the surrounding structure may also be a viral particle, a bacterial cell or a eukaryotic cell. In various embodiments, a composition may comprise cells of Bacillus pumilus or Bacillus subtilis expressing the proteases of the invention, or cell culture supernatants of such cells.

In the context of the present invention, the feature that a protease has the indicated substitutions means that it contains one (of the indicated) substitution(s) at the respective position, i.e., at least the indicated positions are not otherwise mutated or deleted, e.g., by fragmentation of the protease. In preferred embodiments, the proteases described herein have the sequence of SEQ ID NO:1 with the exception of the explicitly mentioned substitutions, i.e., are 100% identical to the sequence according to SEQ ID NO:1 except for the substituted positions.

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 and commonly used in the state of the art (e.g., Altschul et al., Basic local alignment search tool, J. Mol. Biol., 1990, 215, 403-410, and Altschul et al., Gapped BLAST and PSI-BLAST: a new generation of protein database search programs, Nucleic Acids Res., 1997, 25, 3389-3402) and is basically done by assigning similar sequences of nucleotides or amino acids in the nucleic acid or amino acid sequences to each other. A tabular assignment of the corresponding positions is referred to as alignment. Another algorithm available in the state of the art is the FASTA algorithm. Sequence comparisons (alignments), in particular multiple sequence comparisons, are created using computer programs. Frequently used are, e.g., the Clustal series (e.g., Chenna et al., Multiple sequence alignment with the Clustal series of programs, Nucleic Acid Res., 2003, 31 , 3497-3500), T-Coffee (e.g., Notredame et al., T-Coffee: A novel method for multiple sequence alignments, J. Mol. Biol., 2000, 302, 205-217) or programs based on these programs or algorithms. Furthermore, sequence comparisons (alignments) are possible with the computer program Vector NTI® Suite 10.3 (Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California, USA) with the specified standard parameters, whose AlignX module for sequence comparisons is based on ClustalW, or Clone Manager 10 (use of the BLOSUM 62 scoring matrix for sequence alignment at amino acid level). Unless otherwise stated, the sequence identity specified herein is determined using the BLAST algorithm. Such a comparison also allows a statement to be made about the similarity of the compared sequences to each other. It is usually expressed as a percentage of identity, i.e., the proportion of identical nucleotides or amino acid residues at the same positions or at positions corresponding to each other in an alignment. The broader concept of homology includes conserved amino acid exchanges in amino acid sequences, i.e., amino acids with similar chemical activity, as these usually have 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 information can be given 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 matches in the sequences. Such regions often have identical functions. They can be small and comprise only a few nucleotides or amino acids. Such small regions often have essential functions for the overall activity of the protein. It may therefore be useful to refer sequence matches only to individual, possibly small regions. Unless otherwise indicated, however, identity or homology data in the present application refer to the total length of the nucleic acid or amino acid sequence indicated in each case.

In the context of the present invention, the indication 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. Furthermore, the assignment of the positions is based on the mature protein. This assignment is also to be used in particular if the amino acid sequence of a protease according to the invention comprises a higher number of amino acid residues than the protease according to SEQ ID NO:1 . Based on the positions mentioned in the amino acid sequence of the protease according to SEQ ID NO:1 , the modification positions in a protease according to the invention are those which are assigned to these positions in an alignment.

In addition to the amino acid changes described above, proteases according to the invention may have further amino acid changes, in particular amino acid substitutions, insertions or deletions. Such proteases are, e.g., further developed by targeted genetic modification, i.e., by mutagenesis processes, and optimized for specific applications or with respect to specific properties (e.g., with regard to their catalytic activity, stability, etc.). Furthermore, nucleic acids according to the invention can be introduced into recombination approaches and thus used to generate completely new types of proteases or other polypeptides. The aim is to introduce targeted mutations such as substitutions, insertions or deletions into the known molecules, e.g., to improve the cleaning performance of the enzymes according to the invention. In particular, the surface charges and/or isoelectric points of the molecules and thus their interactions with the substrate can be changed for this purpose. For example, the net charge of the enzymes can be altered in order to influence substrate binding, particularly for use in washing and cleaning compositions. Alternatively or additionally, the stability or catalytic activity of the protease can be increased by one or more corresponding mutations, thereby improving its cleaning performance. Advantageous properties of single mutations, e.g., single substitutions, can complement each other. A protease which has already been optimized with respect to certain properties, e.g., with respect to its stability during storage and/or its activity and/or its tolerance with respect to the substrate spectrum, can thus be additionally further developed within the scope of the invention.

The following convention is used to describe substitutions that affect exactly one amino acid position (amino acid substitutions): first the naturally occurring amino acid is given in the form of the internationally used one-letter code, followed by the corresponding sequence position and finally the inserted amino acid. Multiple or alternative substitutions within the same polypeptide chain are separated by slashes. Thus, "130D/V" means that position 130 is mutated to D or V. In the case of insertions, additional amino acids are given after the sequence position. In the case of deletions, the missing amino acid is replaced by a symbol, e.g., an asterisk (*) or a dash (-), or a A is placed before the corresponding position. For example, P9T describes the substitution of threonine for proline at position 9, P9TH describes the insertion of histidine after the amino acid threonine at position 9, and P9* or AP9 describes the deletion of proline at position 9. This nomenclature will be known to those skilled in the art of enzyme technology.

A skilled person is able to produce the corresponding nucleic acids up to complete genes by means of methods generally known today, such as chemical synthesis or the polymerase chain reaction (PCR) in conjunction with standard molecular biological and/or protein chemical methods, using known DNA and/or amino acid sequences. Such methods are known, e.g., from Sambrook, J., Fritsch, E.F. and Maniatis, T. 2001. Molecular cloning: a laboratory manual, 3rd edition Cold Spring Laboratory Press.

A composition according to the invention advantageously contains the protease described herein in an amount of from 2 pg to 20 mg, preferably from 5 pg to 17.5 mg, more preferably from 20 pg to 15 mg and most preferably from 50 pg to 10 mg per g of the composition. In various embodiments, the concentration of the protease (active enzyme) described herein in the composition is >0 to 1 wt.%, preferably 0.0001 or 0.001 to 0.1 wt.% based on the total weight of the composition.

A composition according to the invention increasingly preferably contains the protease in an amount of from 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 washing composition.

In preferred embodiments, a composition according to the invention is characterized in that it contains less than 0.2 wt.% boric acid, based on active substance and total weight of the composition. In more preferred embodiments, a composition according to the invention is characterized in that it contains less than 0.1 wt.% boric acid, based on active substance and total weight of the composition. In even more preferred embodiments, a composition according to the invention is characterized in that is contains less than 0.05 wt.% boric acid, based on active substance and total weight of the composition. In most preferred embodiments, a composition according to the invention is a boric acid- free textile washing composition. In the context of the present invention, "boric acid-free" means that a composition does not contain any boric acid.

A composition according to the invention may comprise one or more reversible protease inhibitors). Such reversible protease inhibitor might be selected from the group consisting of boron acids or salts and derivatives thereof, in particular phenylboronic acid derivatives, and peptide inhibitors like peptide aldehydes. A composition according to the invention may comprise the reversible protease inhibitor(s) in a concentration of 0.1 to 2 wt.%, preferably 0.3 to 1 .5 wt.%, based on active substance and total weight of the composition. If several inhibitors are present, these numbers refer to the total concentration.

In the context of the present invention, "phenylboronic acid derivative" means a compound having the formula (I):

wherein R is hydrogen, a hydroxyl group, a Ci-e alkyl group, a substituted C1-6 alkyl group, a C1-6 alkenyl group or a substituted Ci-e alkenyl group. Preferably, the radical R in the phenylboronic acid derivative is a C1-6 alkyl group and further preferably is -CH3, -CH3CH2 or -CH3CH2CH2. Further preferably, the radical R in the phenylboronic acid derivative is hydrogen. Particularly preferred is the phenylboronic acid derivative 4-formyl-phenylboronic acid (4-FPBA).

In preferred embodiments, a composition according to the invention is substantially free of boron- containing compounds. "Substantially free of boron-containing compounds" in this context means that a composition according to the invention contain less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.% and particularly preferably less than 0.01 wt.%, boron-containing compounds, based on active substance and the total weight of the composition. In particularly preferred embodiments, a composition according to the invention is free of boron-containing compounds, i.e., in particular they do not contain any boric acid and/or phenylboronic acid derivatives.

In preferred embodiments, a composition according to the invention does not contain any reversible protease inhibitors.

The compositions according to the invention may contain one or more surfactants, in particular anionic surfactants, nonionic surfactants and mixtures thereof, but cationic, zwitterionic and/or amphoteric surfactants may also be present. The compositions preferably contain 5 to 70 wt.%, more preferably 5 to 60 wt.% and even more preferably 5 to 50 wt.%, surfactant, based on active substance and total weight of the composition. In preferred embodiments, a composition according to the invention comprises 3 to 35 wt.%, preferably 5 to 20 wt.% surfactants, based on active substance and total weight of the composition.

Suitable anionic surfactants are in particular soaps and those containing sulphate or sulphonate groups with preferably alkali ions as cations. Suitable soaps are preferably the alkali salts of saturated or unsaturated C12-18 fatty acids. Such fatty acids can also be used in an incompletely neutralized form. Useful surfactants of the sulphate type include the salts of the sulphuric acid half esters of C12-18 fatty alcohols and the sulphation products of the nonionic surfactants mentioned with a low degree of ethoxylation. Surfactants of the sulfonate type that can be used include, e.g., C9-14 alkyl benzene sulfonates, alkane sulfonates obtained from C12-18 alkanes, e.g., by sulfochlorination or sulfoxidation with subsequent hydrolysis or neutralization, C12-18 olefin sulfonates formed by the reaction of corresponding monoolefins with sulfur trioxide, mixtures of alkene and hydroxyalkane sulfonates, disulfonates, such as those obtained from C12-18 monoolefins with terminal or internal double bonds by sulfonation with gaseous sulfur trioxide and subsequent alkaline or acidic hydrolysis of the sulfonation products, and a-sulfofatty acid esters (ester sulfonates), which are formed by the sulfonation of fatty acid methyl or ethyl esters, e.g., a-sulfonated methyl esters of hydrogenated coconut, palm kernel or tallow fatty acids.

Preferably, the composition according to the invention contains 2 to 55 wt.%, preferably 3 to 35 wt.%, more preferably 3 to 25 wt.%, most preferably 3 to 10 wt.%, anionic surfactant, based on active substance and total weight of the composition.

In addition to soaps, the compositions according to the invention can contain more than one surfactant, e.g., they can contain one or two additional anionic surfactant(s), they can contain one anionic surfactant and one nonionic surfactant, they can contain two different anionic surfactants and one nonionic surfactant, they can contain one anionic surfactant and two different nonionic surfactants, or any other suitable combination or number of surfactants. In such case, the additional anionic surfactant is preferably selected from alkyl ether sulphates and the nonionic surfactant is selected from fatty alcohol alkoxylates.

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 in particular 9 to 13 carbon atoms. A particularly preferred representative is sodium dodecylbenzyl sulfonate.

Preferred alk(en)yl sulfates are the alkali and in particular the sodium salts of the sulfuric acid half esters of the C12-18 fatty alcohols, e.g., from coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl or stearyl alcohol or the C10-20 oxoalcohols and those half esters of secondary alcohols of these chain lengths. Further preferred are alk(en)yl sulphates of the mentioned chain length, which contain a synthetic, petrochemically produced unbranched alkyl residue, which have an analogous degradation behavior as the adequate compounds based on fatty chemical raw materials. The C12-16 alkyl sulphates and C12-15 alkyl sulphates as well as C14-15 alkyl sulphates are preferred for washing purposes.

The sulphuric acid monoesters of unbranched or branched C7-21 alcohols ethoxylated with 1 to 6 mol ethylene oxide, such as 2-methyl-branched C9-11 alcohols with an average of 3.5 mol ethylene oxide (EO) or C12-18 fatty alcohols with 1 to 4 EO, are also suitable.

Suitable alkyl ether sulphates are, e.g., compounds of the formula

R¹-O-(AO)_n-SO₃- X⁺. In this formula, R¹ is a linear or branched, substituted or unsubstituted alkyl radical, preferably a linear, unsubstituted alkyl radical, particularly preferably a fatty alcohol radical. Preferred radicals R¹ are selected from decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl radicals and mixtures thereof, the representatives with an even number of carbon atoms being preferred. Particularly preferred radicals R¹ are derived from C12-18 fatty alcohols, e.g., from coconut fatty alcohol, tallow fatty alcohol, lauryl, myristyl, cetyl or stearyl alcohol or from C10-20 oxoalcohols. AO stands for an ethylene oxide (EO) or propylene oxide (PO) grouping, preferably for an ethylene oxide grouping. The index n stands for an integer from 1 to 50, preferably from 1 to 20 and in particular from 2 to 10. n is particularly preferably the number 2, 3, 4, 5, 6, 7 or 8. X⁺ stands for a monovalent cation or the n-th part of an n-valent cation, the alkali metal ions being preferred, including Na⁺ or K⁺, with Na⁺ being extremely preferred. Further cations X⁺ can be selected from NHT, % Zn²⁺, % Mg²⁺, % Ca²⁺, % Mn²⁺ and mixtures thereof.

In various embodiments, the alkyl ether sulfate may be selected from fatty alcohol ether sulfates of the formula

where k = 11 to 19, n = 2, 3, 4, 5, 6, 7 or 8. Particularly preferred representatives are Na-Ci2-14 fatty alcohol ether sulfates with 2 EO (k = 1 1-13, n = 2). The stated degree of ethoxylation represents a statistical mean value, which may be an integer or a fractional number for a specific product. The indicated degrees of alkoxylation represent statistical averages, which may be an integer or a fractional number for a specific product. Preferred alkoxylates/ethoxylates have a narrowed homolog distribution (narrow range ethoxylates, NRE).

It has been shown to be advantageous for cold wash performance if the washing compositions also contain soap(s). Preferred washing compositions are therefore characterized in that they contain soap(s). 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 soap mixtures derived in particular from natural fatty acids, e.g., coconut, palm kernel or tallow fatty acids, are suitable.

Suitable nonionic surfactants are in particular alkyl glycosides and ethoxylation and/or propoxylation products of alkyl glycosides or linear or branched alcohols each having 8 to about 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 long-chain alcohol derivatives mentioned with regard to the alkyl moiety, and of alkylphenols with 5 to 12 C atoms in the alkyl radical are also useful.

Advantageously alkoxylated, preferably ethoxylated, in particular primary alcohols with preferably 8 to 18 C atoms and an average of 1 to 12 moles of ethylene oxide (EO) per mole of alcohol are used as nonionic surfactants, in which the alcohol residue can be linear or preferably methyl-branched in the 2-position or can contain linear and methyl-branched residues in the mixture, as they are usually present in oxoalcohol residues. However, alcohol ethoxylates with linear residues from alcohols of native origin with 12 to 18 C atoms, e.g., from coconut, palm, tallow fat or oleyl alcohol, and an average of 2 to 8 EO per mole of alcohol are particularly preferred. Preferred ethoxylated alcohols include, e.g., C12-14 alcohols with 3 EO or 4 EO, C9-11 alcohol with 7 EO, C13-15 alcohols with 3 EO, 5 EO, 7 EO or 8 EO, C12-18 alcohols with 3 EO, 5 EO or 7 EO and mixtures of these, such as mixtures of C12-14 alcohol with 3 EO and C12-18 alcohol with 5 EO. The stated degrees of ethoxylation represent statistical averages, which may be an integer or a fractional number for a specific product. Preferred alcohol ethoxylates have a narrowed homolog distribution (narrow range ethoxylates, NRE). In addition to these nonionic surfactants, fatty alcohols with more than 12 EO can also be used.

Examples of this are tallow fatty alcohols with 14 EO, 25 EO, 30 EO or 40 EO.

A further class of preferably used nonionic surfactants, which are used either as the sole nonionic surfactant or in combination with other nonionic 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.

Another class of nonionic surfactants that can be advantageously used are alkyl polyglycosides (APG). Applicable alkyl polyglycosides have the general formula

RO(G)z, in which R is a linear or branched, in particular methyl-branched in the 2-position, saturated or unsaturated, aliphatic radical with 8 to 22, preferably 12 to 18 C atoms and G is the symbol which stands for a glycose unit with 5 or 6 C atoms, preferably for glucose. The degree of glycosidation z is between 1 and 4, preferably between 1 and 2 and in particular between 1 .1 and 1 .4. Linear alkyl polyglycosides, i.e., alkyl polyglycosides in which the polyglycosyl radical is a glucose radical and the alkyl radical is an n-alkyl radical, are preferably used.

Nonionic surfactants of the amine oxide type, e.g., N-cocoalkyl-N,N-dimethylamine oxide and N- tallowalkyl-N,N-dihydroxyethylamine oxide, and fatty acid alkanolamides may also be suitable. The amount of these nonionic surfactants is preferably not more than that of the ethoxylated fatty alcohols, in particular not more than half thereof.

Suitable amphoteric surfactants are, e.g., betaines of the formula

(Rⁱⁱⁱ)(R^iv)(R^v)N⁺CH₂COO-, in which R'" is an alkyl radical having 8 to 25, preferably 10 to 21 carbon atoms, which may be interrupted by heteroatoms or heteroatom groups, and R^iv and R^v are identical or different alkyl radicals having 1 to 3 carbon atoms, in particular Cio-is-alkyl dimethyl carboxymethyl betaine and C11- 17-alkylamidopropyldimethyl carboxymethyl betaine.

Suitable cationic surfactants include the quaternary ammonium compounds of the formula (Rvi)(Rvii)(Rviii)(Rix)_N+X-, in which R^vi to R^ix stand for four identical or different, in particular two long-chain and two shortchain, alkyl radicals and X- stands for an anion, in particular a halide ion, e.g., didecyldimethylammonium chloride, alkylbenzyldidecylammonium chloride and mixtures thereof. Other suitable cationic surfactants are the quaternary surface-active compounds, in particular with a sulfonium, phosphonium, iodonium or arsonium group, which are also known as antimicrobial agents. By using quaternary surface-active compounds with antimicrobial activity, the composition can be designed with an antimicrobial effect or its antimicrobial effect, which may already be present due to other ingredients, can be improved.

In preferred embodiments, a composition according to the invention comprises, in each case based on active substance and total weight of the composition, 2 to 20 wt.%, preferably 3 to 17 wt.%, anionic surfactants, 1 to 10 wt.%, preferably 3 to 8 wt.%, nonionic surfactants, 0 to 2 wt.%, preferably 1 to 1 .75 wt.%, soap, and 0 to 5 wt.%, preferably 0 to 3 wt.% fatty acids.

In preferred embodiments, a composition according to the invention comprises, in each case based on active substance and total weight of the composition, FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%.

Washing or cleaning compositions according to the invention may exclusively contain a protease as described herein. Alternatively, they may also contain further hydrolytic enzymes or other enzymes in a concentration appropriate for the effectiveness of the composition. Thus, compositions further comprising one or more further enzymes represent a further embodiment of the invention. Preferred further enzymes are all enzymes which can develop catalytic activity in the composition according to the invention, in particular a lipase, amylase, cellulase, hemicellulase, mannanase, tannase, xylanase, xanthanase, xyloglucanase, B-glucosidase, pectinase, carrageenase, perhydrolase, oxidase, oxidoreductase or other proteases - distinguishable from the proteases described herein - and mixtures thereof. Further enzymes are advantageously each contained in the composition in an amount of 1 x 10^-8 to 5 wt.% based on active protein. Increasingly preferably, each further enzyme is present in an amount of from 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.% in compositions according to the invention, based on active protein and total weight of the composition.

Particularly preferably, the enzymes show synergistic cleaning performance against certain soiling or stains, i.e., the enzymes contained in the composition support each other in their cleaning performance. Such synergism is particularly preferred between the protease contained according to the invention and another enzyme of a composition according to the invention. Synergistic effects can occur not only between different enzymes, but also between one or more enzymes and further ingredients of the composition according to the invention.

Preferred textile washing compositions according to the invention comprise at least one protease and at least one amylase. In a further preferred embodiment of the invention, textile washing compositions comprise at least one protease and at least one cellulase. In a further preferred embodiment, textile washing compositions comprise at least one protease and at least one lipase. In a further preferred embodiment, textile washing compositions comprise at least one protease, at least one amylase and at least one lipase. In a further preferred embodiment, textile washing compositions comprise at least one protease, at least one amylase and at least one cellulase. In a further preferred embodiment, textile washing compositions comprise at least one protease, at least one amylase, at least one cellulase and at least one lipase. Textile detergents which contain 3 to 10 different enzymes are particularly preferred, whereby textile detergents which contain 3 to 10 different types of enzymes can be of particular preference with regard to cleaning performance over a very large stain spectrum.

In preferred embodiments, a composition according to the invention comprises at least one enzyme and increasingly preferably at least two, three, four or five enzymes, preferably selected from the group consisting of amylases, proteases, lipases, cellulases, mannanases and mixtures thereof, in a total amount of from 0.01 to 10 wt., preferably from 0.1 to 8 wt., more preferably from 0.2 to 6 wt.%, in each case based on active protein and total weight of the composition.

Examples of proteases which can be used in addition to the proteases described herein in compositions according to the invention are the subtilisins BPN' from Bacillus amyloliquefaciens and Carlsberg from 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 subtilisins but no longer as subtilisins in the narrower sense. Subtilisin Carlsberg is available in a further developed form under the trade name Alcalase® from the company Novozymes. Subtilisins 147 and 309 are marketed by Novozymes under the trade names Esperase® and Savinase® respectively. Protease variants are derived from the protease from Bacillus lentus DSM 5483, described in e.g., WO 95/23221 , WO 92/21760 WO 2013/060621 and EP 3660151 . Other useful proteases are, e.g., the proteases marketed under the trade names Durazym®, Relase®, Everlase®, Nafizym®, Kannase®, Progress Uno 101 L® and Ovozyme® by the company Novozymes, the proteases marketed under the trade names Purafect®, Purafect® OxP, Purafect® Prime, Excellase®, Properase®, Preferenz P100® and Preferenz P300® from the company Danisco/IFF, which is marketed under the trade name Lavergy pro 104 LS® from the company BASF, which is marketed under the trade name Protosol® from the company Advanced Biochemicals Ltd., under the trade name Wuxi® from the company Wuxi Snyder Bioproducts Ltd., the enzymes available under the trade names Proleather® and Protease P® from the company Amano Pharmaceuticals Ltd. and the enzyme available under the name Proteinase K-16 from the company Kao Corp. The proteases from Bacillus gibsonii and Bacillus pumilus, which are disclosed in WO 2008/086916, WO 2007/131656, WO 2017/215925, WO 2021/175696 and WO 2021/175697, are also particularly preferably used. Other proteases that can be used are those that are naturally present in the microorganisms Stenotrophomonas maltophilia, in particular Stenotrophomonas maltophilia K279a, Bacillus intermedius and Bacillus sphaericus.

Examples of amylases are the a-amylases from Bacillus licheniformis, Bacillus amyloliquefaciens or Bacillus stearothermophilus and, in particular, their further developments improved for use in washing or cleaning compositions. The enzyme from Bacillus licheniformis is available from the company Novozymes under the name Termamyl® and from the company Danisco/IFF under the name Purastar® ST. Further development products of this a-amylase are available under the trade names Duramyl® and Termamyl® ultra (both from Novozymes), Purastar® OxAm (Danisco/IFF) and Keistase® (Daiwa Seiko Inc.). The a-amylase from Bacillus amyloliquefaciens is marketed by the company Novozymes under the name BAN®, and derived variants of the a-amylase from Bacillus stearothermophilus under the names BSG® and Novamyl®, also from the company Novozymes. Furthermore, the a-amylase from Bacillus sp. A 7-7 (DSM 12368) and the cyclodextrin glucanotransferase (CGTase) from Bacillus agaradherens (DSM 9948) should be emphasized for this purpose. Fusion products of all the molecules mentioned can also be used. In addition, the further developments of a-amylase from Aspergillus niger and A. oryzae available under the trade name Fungamyl® from the company Novozymes are suitable. Other commercial products that are advantageously usable include Amylase-LT® and Stainzyme® or Stainzyme® ultra or Stainzyme® plus as well as Amplify™ 12L, Amplify Prime™ 120L or Amplify Prime™ 100L, the latter also from Novozymes, and the PREFERENZ S® series from Danisco/IFF, including e.g., PREFERENZ S100®, PREFERENZ S1000® or PREFERENZ S210®. Variants of these enzymes obtained by point mutations can also be used in compositions according to the invention.

Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases are cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulases from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum, which are disclosed in US 4435307, US 5648263, US 5691178, US 5776757 and WO 89/09259. Particularly suitable cellulases are the alkaline or neutral cellulases with color care properties. Examples of such cellulases are cellulases described in EP 0495257, EP 0531372, WO 96/11262, WO 96/29397 and WO 98/08940. Examples of cellulases having endo-1 ,4-glucanase activity (EC 3.2.1 .4) are described in WO 2002/099091 , e.g., those having a sequence of at least 97% identity to the amino acid sequence of positions 1 to 773 of SEQ ID NO:2 of WO 2002/099091 . Another example may comprise a GH44 xyloglucanase, e.g., a xyloglucanase enzyme having a sequence of at least 60% identity to positions 40 to 559 of SEQ ID NO:2 of WO 2001/062903. Commercially available cellulases include e.g., Celluzyme™, Carezyme™, Carezyme Premium™, Celluclean™ (e.g., Celluclean™ 5000L and Celluclean™ 4000T), Celluclean Classic™, Cellusoft™, Endolase®, Renozyme® and Whitezyme™ (Novozymes A/S), Clazinase™ and Puradax HA™ (Danisco/IFF), KAC-500(B)™ (Kao Corporation), Revitalenz™ 1000, Revitalenz™ 2000 and Revitalenz™ 3000 (Danisco/IFF), as well as Ecostone® and Biotouch® (AB Enzymes).

Other enzymes that can be used include lipases or cutinases, in particular because of their triglyceride-cleaving activities, but also to produce peracids in situ from suitable precursors. Suitable lipases and cutinases are those of bacterial or fungal origin. Chemically modified or protein- engineered mutant enzymes are included. Examples include lipase from Thermomyces, e.g., from T. lanuginosus (formerly called Humicola lanuginosa) as described in EP 0258068 and EP 0305216, cutinase from Humicola, e.g., H. insolens (WO 96/13580), lipase from strains of Pseudomonas (some now renamed Burkholderia), e.g., P. alcaligenes or P. pseudoalcaligenes, P. cepacia, P. sp. strain SD705, P. wisconsinensis, Streptomyces lipases of the GDSL type, cutinase from Magnaporthe grisea, cutinase from Pseudomonas mendocina, lipase from Thermobifida fusca, lipase from Geobacillus stearothermophilus, lipase from Bacillus subtilis and lipase from Streptomyces griseus and S. pristinaespiralis. Preferred lipases include, e.g., lipases originally obtainable from Humicola lanuginosa (Thermomyces lanuginosus) or further developed therefrom, in particular those with one or more of the following amino acid substitutions starting from said lipase in positions D96L, T213R and/or N233R, particularly preferably T213R and N233R. Preferred commercial lipase products include Lipolase™, Lipex™, Lipolex™ and Lipoclean™ (Novozymes A/S), Lumafast (Danisco/IFF) and Lipomax (Gist-Brocades).

Suitable mannanases include Bacillus subtilis endo-p-mannanase, Bacillus sp. 1633 endo- - mannanase, Bacillus sp. AAI12 endo-p-mannanase, Bacillus sp. AA349 endo-p-mannanase, Bacillus agaradhaerens NCIMB 40482 endo-p-mannanase, Bacillus halodurans endo- -mannanase, Bacillus clausii endo-p-mannanase, Bacillus licheniformis endo-p-mannanase, Humicola insolens endo-p- mannanase and Caldocellulosiruptor sp. endo-p-mannanase (e.g., US 6060299, WO 99/64573, US 6566114 and WO 99/64619).

Pectate lyases suitable for washing and cleaning compositions are described, e.g., in WO 2003/095638 or WO 2015/121133. Examples of suitable pectinolytic enzymes are also those sold under the trade names Gamanase®, Pektinex AR®, X-Pect® or Pectaway® by the company Novozymes, under the trade names Rohapect UF®, Rohapect TPL®, Rohapect PTE100®, Rohapect MPE®, Rohapect MA plus HC, Rohapect DA12L®, Rohapect 10L®, Rohapect B1 L® from the company AB Enzymes and under the trade name Pyrolase® from the company Diversa Corp, enzymes and enzyme preparations.

According to the invention, 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 to increase the bleaching effect. Advantageously, organic, particularly preferably aromatic, compounds interacting with the enzymes are also preferably added in order to increase the activity of the respective oxidoreductases (enhancers) or to ensure the flow of electrons (mediators) if the redox potentials between the oxidizing enzymes and the soiling are very different.

In washing or cleaning compositions according to the invention, the enzymes to be used may also be formulated together with additives, e.g., from fermentation. In liquid formulations, the enzymes are preferably used as enzyme liquid formulation(s).

The enzymes are generally not provided in the form of the pure protein, but in the form of stabilized, storable and/or transportable preparations. These pre-formulated preparations include, e.g., solid preparations obtained by granulation, extrusion or lyophilization, or, particularly in the case of liquid or gel-like compositions, solutions of the enzymes, preferably as concentrated as possible, low in water and/or mixed with stabilizers or other auxiliaries.

Alternatively, the enzymes can be encapsulated for both the solid and liquid dosage forms, e.g., by spray drying or extrusion of the enzyme solution together with a preferably natural polymer or in the form of capsules, e.g., those in which the enzymes are enclosed as in a solidified gel or in those of the core-shell type, in which an enzyme-containing core is coated with a protective layer that is impermeable to water, air and/or chemicals. Additional active ingredients, e.g., stabilizers, emulsifiers, pigments, bleaching agents or dyes, can also be applied in superimposed layers. Such capsules are applied by methods known per se, e.g., by shaking or rolling granulation or in fluid-bed processes. Advantageously, such granules, e.g., by applying polymeric film formers, are low-dust and stable in storage due to the coating.

Furthermore, it is possible to formulate two or more enzymes together so that a single granule has several enzyme activities.

The enzymes can also be incorporated into water-soluble films, such as those used in the packaging of washing or cleaning compositions in unit dose form. Such a film allows the enzymes to be released upon contact with water. As used herein, "water-soluble" refers to a film structure which is preferably completely water-soluble. Preferably, such a film comprises (fully or partially hydrolyzed) polyvinyl alcohol (PVA).

Washing and cleaning performance is understood to mean the ability of a washing or cleaning composition to partially or completely remove an existing stain, in particular the brightening performance on one or more stains on textiles. Examples of such stains are blood on cotton or chocolate milk/carbon black on cotton, cocoa on cotton or porridge on cotton. In the context of the invention, both the washing or cleaning composition comprising the protease or the washing or cleaning liquor formed by this composition and the protease itself have a respective cleaning performance. The cleaning performance of the protease thus contributes to the cleaning performance of the composition or the washing or cleaning liquor formed by the composition.

The washing or cleaning liquor is understood to be the ready-to-use solution containing the washing or cleaning composition which acts on textiles or fabrics or hard surfaces, in particular dishes, and thus comes into contact with the stains present on the textiles or fabrics or hard surfaces, in particular dishes. Usually, the washing or cleaning liquor is formed when the washing or cleaning process begins and the washing or cleaning composition is diluted with water, e.g., in a washing machine or dishwasher or in another suitable container.

The cleaning performance can be determined in a washing system containing a washing composition in a dosage of between 2.0 and 8.0 grams per liter of washing liquor and the protease. The proteases to be compared are used in equal concentrations (based on active protein). The use of enzymes with the same activity ensures that the enzymatic properties, e.g., the cleaning performance on certain stains, can be compared even if the ratio of active substance to total protein (the specific activity values) differs. In general, a low specific activity can be compensated by adding a higher amount of protein. Furthermore, the enzymes to be tested can be used in the same amount or weight even if the enzymes to be tested have different affinities for the test substrate in an activity assay. In this context, the term "same amount of substance" refers to the use of the enzymes to be tested in the same molar amount. The term "same weight" refers to the same weight of the enzymes to be tested.

The concentration of the protease in the washing composition intended for such a washing system is 0.001 to 0.1 wt.%, preferably 0.01 to 0.06 wt.%, based on active protein and total weight of the composition.

A liquid reference detergent for such a washing system may, e.g., be composed as follows (all figures in percent by weight (wt.%), based on active substance and total weight of the composition): 4.4% alkylbenzene sulfonic acid, 5.6% other anionic surfactants, 2.4% C12-18 Na salts of fatty acids (soaps), 4.4% nonionic surfactants, 0.2% phosphonates, 1.4% citric acid, 0.95% NaOH, 0.01% defoamer, 2% glycerol, 0.08% preservatives, 1 % ethanol, remainder demineralized water. Preferably, the dosage of the liquid washing composition is between 4.5 and 6.0 grams per liter of washing liquor, e.g., 4.7, 4.9 or 5.9 grams per liter of washing liquor. Washing is preferably carried out in a pH value range between pH 7 and pH 10.5, preferably between pH 8 and pH 9.

The cleaning performance on a stain on cotton is determined by measuring the degree of cleaning of the washed textiles. For example, the washing process may be carried out for 60 minutes at a temperature of about 20°C or about 40°C and the water may have a water hardness of between 15.5°dH and 16.5°dH (German hardness). In the context of the invention, the cleaning performance is determined, e.g., at 20°C or 40°C using a liquid washing composition as disclosed herein, the washing process preferably being carried out for 60 minutes at 600 rpm.

The degree of whiteness, i.e., the brightening of the stains, as a measure of the cleaning performance is determined by optical measuring methods, preferably photometrically. A suitable device is the Minolta CM508d spectrometer. The devices used for measurement are usually calibrated beforehand with a white standard, preferably a white standard supplied with the device.

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

Methods for determining protease activity are well-known to those skilled in the art in the field of enzyme technology and are routinely used by them. For example, such methods are disclosed in Tenside, Vol. 7, 1970, pp. 125-132. Alternatively, the 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 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 measuring time is 5 min and the measuring interval is 20 s to 60 s. The protease activity is usually expressed in protease units (PE). Suitable protease activities are e.g., 2.25, 5 or 10 PE per ml of wash liquor. However, protease activity is not zero.

An alternative test for determining the proteolytic activity of the proteases used according to the invention is an optical measurement method, preferably a photometric method. The test suitable for this purpose comprises the protease-dependent cleavage of the substrate protein casein. This is cleaved by the protease into a large number of smaller partial products. The totality of these partial products exhibits an increased absorbance at 290 nm compared to non-cleaved casein, whereby this increased absorbance can be determined using a photometer and thus a conclusion can be drawn about the enzymatic activity of the protease.

The protein concentration can be determined using known methods, e.g., the BCA method (bicinchoninic acid; 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the Biuret method (Gornall et al., J. Biol. Chem. 177, 1948, 751-766). The active protein concentration can be determined in this respect by titration of the active centers using a suitable irreversible inhibitor and determination of the residual activity (see Bender et al., J. Am. Chem. Soc. 1966, 88, 24, 5890-5913).

According to the invention, a washing or cleaning composition is to be understood to mean all conceivable types of washing or cleaning compositions, both concentrates and undiluted compositions, for use on a commercial scale, in the washing machine or for hand washing or cleaning. This includes, e.g., washing compositions for textiles, carpets or natural fibers for which the term washing composition or detergent is used. This also includes, e.g., dishwashing composition for dishwashers (automatic dishwashing agents) or manual dishwashing agents or cleaners for hard surfaces such as metal, glass, porcelain, ceramics, tiles, stone, painted surfaces, plastics, wood or leather, for which the term cleaning composition is used, i.e., in addition to manual and automatic dishwashing agents, also scouring agents, glass cleaners, toilet scent rinsers, etc. Washing and cleaning compositions within the scope of the invention also include washing aids which are added to the actual washing composition during manual or mechanical textile washing in order to achieve a further effect. Furthermore, washing and cleaning compositions within the scope of the invention also include textile pre- and post-treatment agents, i.e., agents with which the laundry item is brought into contact before the actual washing, e.g., for dissolving stubborn soiling, and also agents which, in a step subsequent to the actual textile washing, impart further desirable properties to the laundry item, such as a pleasant feel, crease resistance or low static charge. The latter agents include fabric softeners.

Embodiments of the present invention include all solid, powdery, liquid, gel-like or pasty dosage forms of compositions according to the invention, which may also consist of several phases and may be present in compressed or non-compressed form. The composition may be present as a free- flowing powder, in particular with a bulk density of 300 g/l to 1200 g/l, in particular 500 g/l to 900 g/l or 600 g/l to 850 g/l. Solid dosage forms of the composition also include extrudates, granules, tablets or pouches. Alternatively, the composition may also be liquid, gel or paste, e.g., in the form of a nonaqueous liquid detergent or a non-aqueous paste or in the form of an aqueous liquid detergent or an aqueous paste. Liquid compositions are generally preferred. Furthermore, the composition may be present as a single component system. Such compositions consist of one phase. Alternatively, a composition can also consist of several phases. Such a composition is thus divided into several components.

In a preferred embodiment, the composition according to the invention is a textile washing composition.

In a preferred embodiment, the composition according to the invention is a liquid textile washing composition.

In a preferred embodiment, the composition according to the invention is a pre-portioned laundry detergent, in particular a laundry detergent portion unit comprising a laundry detergent preparation according to the invention and a water-soluble film which completely encloses the laundry detergent preparation. The water-soluble film in which the detergent composition is packaged may comprise one or more structurally different water-soluble polymer(s). Polymers from the group of (possibly acetalized) polyvinyl alcohols (PVAL) and their copolymers are particularly suitable as water-soluble polymer(s). Water-soluble films are preferably based on a polyvinyl alcohol or a polyvinyl alcohol copolymer whose molecular weight is in the range from 10000 to 1000000 g/mol, preferably from 20000 to 500000 g/mol, particularly preferably from 30000 to 100000 g/mol and especially from 40000 to 80000 g/mol. Suitable water-soluble films for use are marketed by the company MonoSol LLC, e.g., under the designation M8630, M8720, M8310, C8400 or M8900. Also suitable are films with the designation Solublon® PT, Solublon® GA, Solublon® KC or Solublon® KL from Aicello Chemical Europe GmbH or the VF-HP films from Kuraray.

If the compositions according to the invention are present 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 the total weight of the composition.

The washing or cleaning compositions according to the invention, which may be present as powdery or granular solids, in compacted or post-compacted particle form, as homogeneous solutions or suspensions, may contain, in addition to a protease described herein, all known ingredients commonly used in such compositions, at least one further ingredient preferably being present in the composition. The compositions according to the invention may in particular contain surfactants, builders, polymers, glass corrosion inhibitors, corrosion inhibitors, bleaching agents such as peroxygen compounds, bleach activators or bleach catalysts. They may also contain water-miscible organic solvents, other enzymes, enzyme stabilizers, sequestering agents, electrolytes, pH regulators and/or other auxiliaries such as optical brighteners, greying inhibitors, colour transfer inhibitors, foam regulators, colourants and fragrances, as well as combinations thereof. Advantageous ingredients of compositions according to the invention are disclosed in e.g., WO 2009/121725, beginning on page 5, penultimate paragraph, and ending on page 13 after the second paragraph. This disclosure is expressly referred to and the content of the disclosure therein is included in the present patent application.

Another preferred component of washing compositions according to the invention are complexing agents. Particularly preferred complexing agents are the phosphonates, provided that their use is permitted by regulation. In addition to 1-hydroxyethane-1 ,1-diphosphonic acid, the complexing phosphonates include a number of different compounds, such as diethylene triamine penta- (methylene phosphonic acid) (DTPMP). Hydroxyalkane or aminoalkane phosphonates are particularly preferred in this application. 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, whereby the disodium salt reacts neutrally and the tetrasodium salt alkaline (pH 9). The preferred aminoalkane phosphonates are ethylene diamine tetramethylene phosphonate (EDTMP), diethylene triamine pentamethylene phosphonate (DTPMP) and their higher homologs. They are preferably used in the form of the neutrally reacting sodium salts, e.g., as hexasodium salt of EDTMP or as hepta- and octa-sodium salt of DTPMP. HEDP from the class of phosphonates is the preferred builder. The aminoalkane phosphonates also have a pronounced heavy metal binding capacity. Accordingly, especially if the compositions also contain bleach, it may be preferable to use aminoalkane phosphonates, in particular DTPMP, or to use mixtures of the aforementioned phosphonates. A washing composition according to the invention may contain one or more phosphonate(s) from the group amino trimethylene phosphonic acid (ATMP) and/or salts thereof; ethylene diamine tetra- (methylene phosphonic acid) (EDTMP) and/or salts thereof; diethylene triamine penta-(methylene phosphonic acid) (DTPMP) and/or salts thereof; 1-hydroxyethane-1 ,1-diphosphonic acid (HEDP) and/or salts thereof; 2-phosphonobutane-1 ,2,4-tricarboxylic acid (PBTC) and/or salts thereof; hexamethylene diamine tetra-(methylene phosphonic acid) (HDTMP) and/or salts thereof; nitrilotri- (methylene phosphonic acid) (NTMP) and/or salts thereof. Washing compositions which contain 1- hydroxyethane-1 ,1-diphosphonic acid (HEDP) or diethylene triamine penta-(methylene phosphonic acid) (DTPMP) as phosphonates are particularly preferred. Of course, the washing compositions according to the invention may contain two or more different phosphonates.

Washing compositions according to the invention may contain at least one complexing agent from the group of phosphonates, preferably 1-hydroxyethane-1 ,1 -diphosphonate, preferably in an amount of 0.1 to 8.0 wt.%, more preferably 0.2 to 5.0 wt.%, further preferably 0.3 to 3.0 wt.% and particularly preferably 0.5 to 2.0 wt.%, based on active substance and total weight of the composition.

In preferred embodiments, a composition according to the invention are substantially free of phosphonate-containing compounds. "Substantially free of phosphonate-containing compounds" in this context means that a composition according to the invention contain less than 2 wt.%, preferably less than 1 wt.%, more preferably less than 0.5 wt.% and particularly preferably less than 0.1 wt.%, phosphonate-containing compounds, based on the total weight of the composition. In particularly preferred embodiments, these compositions are free of phosphonate-containing compounds.

The washing compositions according to the invention further preferably contain builders, preferably at least one water-soluble and/or water-insoluble, organic and/or inorganic builder. The builders include, in particular, silicates, carbonates and organic cobuilders.

Organic cobuilders include, in particular, polycarboxylates/polycarboxylic acids, polymeric polycarboxylates, aspartic acid, polyacetals, dextrins, other organic cobuilders and phosphonates. These substance classes are described below. Organic cobuilder substances may, if desired, be present in amounts of up to 40 wt.%, in particular up to 25 wt.% and preferably from 1 to 8 wt.%, based on active substance and total weight of the composition. Useful organic framework substances are, e.g., the 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 those carboxylic acids which 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 carboxymethyl inulinulines, monomeric and polymeric amino polycarboxylic acids, in particular glycine diacetic acid, methylglycine diacetic acid, glutamine diacetic acid, nitrilotriacetic acid (NTA), iminodisuccinate such as ethylenediamine-N,N'- disuccinic acid and hydroxyamino disuccinates, ethylene diamine tetraacetic acid and polyaspartic acid, polyphosphonic acids, in particular aminotris-(methylene phosphonic acid), ethylene diamine tetrakis-(methylene phosphonic acid), lysine tetra-(methylene phosphonic acid) and 1-hydroxyethane- 1 ,1-diphosphonic acid, polymeric hydroxy compounds such as dextrin and polymeric (poly)carboxylic acids, in particular those accessible by oxidation of polysaccharides or dextrins, and/or polymeric acrylic acids, methacrylic acids, maleic acids and mixed polymers of these, which may also contain small amounts of polymerizable substances without carboxylic acid functionality in polymerized form. If desired, such organic builder substances can be present in quantities of up to 50 wt.%, in particular up to 25 wt.%, preferably from 10 to 20 wt.% and particularly preferably from 1 to 5 wt.%, based on active substance and total weight of the composition.

In addition to their builder effect, the free acids typically also have the property of an acidifying component and thus also serve to adjust a lower and milder pH value of washing compositions. In particular, citric acid, succinic acid, glutaric acid, adipic acid, gluconic acid and any mixtures of these should be mentioned. Citric acid or salts of citric acid are used with particular preference as a framework substance. Further particularly preferred backbone substances are selected from methyl glycine diacetic acid (MGDA), glutamic acid diacetate (GLDA), aspartic acid diacetate (ASDA), hydroxy ethyl imino diacetate (HEIDA), imino disuccinate (IDS) and ethylene diamine disuccinate (EDDS), carboxy methyl inulin and polyaspartate. In preferred embodiments, citric acid and/or citrate is used as a water-soluble organic builder.

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, even more preferably sodium citrate, based on active substance and total weight of the composition. Citric acid/citrate can each be used in the form of their hydrates, e.g., citric acid can be used in the form of the monohydrate, citrate in the form of the trisodium citrate dihydrate.

In preferred embodiments, the builder substances may be selected from MGDA and GLDA. As used herein, the term "MGDA" includes, but is not limited to, methyl glycine diacetic acid, a-alanine diacetic acid, N-(1 -carboxy ethyl)-imino diacetic acid and N,N-bis(carboxy methyl)-DL-alanine, wherein the free acid forms and the corresponding salts, preferably alkali salts, in particular trisodium salts, are included. As used herein, the term "GLDA" includes, but is not limited to, glutamic acid-diacetic acid, L-glutamic acid-N,N-diacetic acid and N,N-bis(carboxylatomethyl)-L-glutamate, wherein free acid forms and corresponding salts, preferably alkali metal salts, in particular tetrasodium salts, are included. Although, based on the total weight of the composition, higher MGDA or GLDA concentrations are possible, in particular up to 25 wt.%, 0.2 to 5 wt.%, preferably 0.25 to 3 wt.%, even more preferably 0.5 to 2 wt.%, MGDA, preferably MGDA trisodium salt (MGDA-Nas) is preferred. When using GLDA as builder, the use, based on the total weight of the composition, 0.2 to 5 wt.%, preferably 0.25 to 3 wt.%, even more preferably 0.5 to 2 wt.%, GLDA, preferably GLDA tetrasodium salt (GLDA-N34) is preferred.

Polymeric polycarboxylates are also suitable as builders, these are, e.g., the alkali metal salts of polyacrylic acid or polymethacrylic acid, e.g., those with a relative molecular mass of 500 to 70000 g/mol. For the purposes of this application, the molecular masses indicated for polymeric polycarboxylates are weight-average molecular masses Mw of the respective acid form, which were basically determined by gel permeation chromatography (GPC), using a UV detector. The measurement was carried out against an external polyacrylic acid standard, which provides realistic molar weight values due to its structural relationship with the polymers under investigation. This data differs significantly from the molar weight data in which polystyrene sulfonic acids are used as a standard. The molecular weights measured against polystyrene sulfonic acids are generally significantly higher than the molecular weights stated in this application. Suitable polymers are in particular polyacrylates, which preferably have a molecular mass of 2000 to 20000 g/mol. Due to their superior solubility, the short-chain polyacrylates from this group, which have molar masses of 2000 to 10000 g/mol, and particularly preferably 3000 to 5000 g/mol, may again be preferred. Also suitable are copolymeric polycarboxylates, in particular those of acrylic acid with methacrylic acid and of acrylic acid or methacrylic acid with maleic acid. Copolymers of acrylic acid with maleic acid which contain 50 to 90 wt.% acrylic acid and 50 to 10 wt.% maleic acid have proved to be particularly suitable. Their relative molecular mass, based on free acids, is generally 2000 to 70000 g/mol, preferably 20000 to 50000 g/mol and in particular 30000 to 40000 g/mol.

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 above-mentioned organic builder substances.

In addition to the above-mentioned water-soluble organic builders, the compositions of the invention may also contain inorganic water-soluble builders. In particular, alkali silicates, alkali carbonates, alkali hydrogen barbonates, alkali phosphates and/or sesquicarbonates, which may be present in the form of their alkaline, neutral or acidic sodium or potassium salts, may be considered as water-soluble inorganic builder materials. Small amounts of calcium carbonates may also be contained in solid textile detergents. Water-soluble crystalline and/or amorphous alkali silicates, e.g., are suitable. The alkali silicates which can be used as builders in the compositions according to the invention preferably have a molar ratio of alkali oxide to SiC>2 below 0.95, in particular from 1 :1.1 to 1 :12, and can be amorphous or crystalline. Preferred alkali silicates are the sodium silicates, in particular the amorphous sodium silicates, with a molar ratio Na2O:SiC>2 of 1 :2 to 1 :2.8. Crystalline layered silicates of the general formula Na2Si_xC>2x+i • y H2O, in which x, the so-called modulus, is a number from 1 .9 to 22, in particular 1 .9 to 4, and y is a number from 0 to 33, and preferred values for x are 2, 3 or 4, are preferably used as crystalline silicates, which can be present alone or in a mixture with amorphous silicates. Preferred crystalline layered silicates are those in which x in the general formula mentioned assumes the values 2 or 3. In particular, both B- and 6-sodium disilicates (Na2Si2C>5 • y H2O) are preferred. Practically anhydrous crystalline alkali metal silicates produced from amorphous alkali metal silicates of the above-mentioned general formula, in which x is a number from 1 .9 to 2.1 , can also be used in compositions according to the invention. In a further embodiment of compositions 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 from 1 .9 to 3.5 are used in a further embodiment of compositions 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 in particular 1 :1 to 2:1 . Crystalline layered silicates of the above formula are marketed by Clariant GmbH under the trade name Na-SKS. In one embodiment of compositions according to the invention, a granular compound of crystalline phyllosilicate and citrate, of crystalline phyllosilicate and the above-mentioned (co- )polymeric polycarboxylic acid, or of alkali silicate and alkali carbonate, such as is commercially available under the name Nabion® 15, is used. 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, the carbonates (and hyodrogencarbonates), in particular sodium carbonate, and the phosphonic acids/phosphonates are also of importance as water-soluble inorganic builder substances.

The compositions according to the invention are preferably free of phosphate builder, i.e., contain less than 1 wt.%, preferably no intentionally added phosphate builder. In preferred embodiments, a composition according to the invention are essentially free of phosphate-containing compounds. "Substantially free of phosphate-containing compounds" in this context means that a composition according to the invention contain less than 2 wt.%, preferably less than 1 wt.%, more preferably less than 0.5 wt.% and particularly preferably less than 0.1 wt.%, phosphate-containing compounds, based on the total weight of the composition. In particularly preferred embodiments, these compositions are free of phosphate-containing compounds.

The compositions may also contain water-insoluble builder substances. The water-insoluble inorganic builder materials used are in particular crystalline or amorphous water-dispersible alkali metal aluminosilicates in amounts of up to 50 wt.%, preferably not more than 40 wt.%, in particular from 3 to 20 wt.% and particularly preferably from 1 to 15 wt.%. Among these, the crystalline sodium aluminosilicates in detergent quality, in particular zeolite A, zeolite P, zeolite MAP and possibly 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 S.p.A.), are preferred. Quantities close to the upper limit mentioned are preferably used in solid, particulate compositions. Suitable aluminosilicates have in particular no particles with a particle size above 30 pm and preferably consist of at least 80 wt.% particles with a size below 10 pm. Their calcium binding capacity, which can be determined according to DE 2412837 A1 , is generally in the range of 100 to 200 mg CaO per gram.

In preferred embodiments, a composition according to the invention comprises a builder system comprising at least one builder, preferably in an amount of 0.5 to 50 wt.%, preferably 0.5 to 20 wt.%, particularly preferably 0.5 to 10 wt.%, based on active substance and total weight of the composition, wherein the builder is selected from the group consisting of polycarboxylic acids such as citric acid, adipic acid, succinic acid, glutaric acid, malic acid, tartaric acid, tartaric acid, tartaric acid and tartaric acid, maleic acid, fumaric acid, sugar acids and carboxymethyl inulinulines or their salts, monomeric and polymeric amino polycarboxylic acids such as glycine diacetic acid, methyl glycine diacetic acid (MGDA), glutamic acid diacetic acid (GLDA), nitrile triacetic acid, imino disuccinate such as ethylene diamine-N,N'-disuccinic acid and hydroxy imino disuccinates, ethylene diamine tetraacetic acid and polyaspartic acid or salts thereof, polyphosphonic acids such as aminotris-(methylene phosphonic acid), ethylene diamine tetrakis-(methylene phosphonic acid), lysine tetra-(methylene phosphonic acid), diethylene triamine penta-(methylene phosphonic acid) (DTPMP) and 1-hydroxyethane-1 ,1- diphosphonic acid (HEDP) or salts thereof, polymeric hydroxy compounds such as dextrin, and mixtures thereof.

In preferred embodiments, a composition according to the invention, comprises, in each case based on active substance and total weight of the composition, 0 to 10 wt.%, preferably 1 to 4 wt.%, citric acid and/or citrate, preferably alkali metal citrate, 0 to 40 wt.%, preferably 0 to 15 wt.%, more preferably 0 to 3 wt.%, alkali metal carbonate, preferably sodium carbonate, 0 to 20 wt.%, preferably 0 to 10 wt.%, alkali metal silicate, 0 to 10 wt.%, preferably 0.5 to 2 wt.%, phosphonic acid and/or alkali metal phosphonate, more preferably HEDP and/or DTPMP, and/or 0 to 10 wt.%, preferably 0 to 3 wt.%, amino polycarboxylic acids, preferably MGDA and/or GLDA.

In addition to the builders described above, the washing composition may contain cleaning-active polymers. The weight proportion of the cleaning-active polymers in the total weight of washing compositions according to the invention is preferably 0.1 to 20 wt.%, preferably 1 .0 to 15 wt.% and further preferably 2.0 to 12 wt.%.

Suitable anti-graying agents or soil release polymers are cellulose ethers, such as carboxy methyl cellulose, methyl cellulose, hydroxyalkyl celluloses and mixed cellulose ethers, such as methyl hydroxyethyl cellulose, methyl hydroxypropyl cellulose and methyl carboxy methyl cellulose. Preferably, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose and mixtures thereof and optionally mixtures thereof with methylcellulose are used. Commonly used soil-release active ingredients include copolyesters containing dicarboxylic acid units, alkylene glycol units and polyalkylene glycol units. The amount of graying inhibitors and/or soil-release agents in compositions according to the invention generally does not exceed 2 wt.% and is preferably 0.5 to 1 .5 wt.%, particularly preferably 0.5 to 2 wt.%.

As optical brighteners for textiles made of cellulose fibers (e.g., cotton) in particular, derivatives of diaminostilbene disulfonic acid or its alkali metal salts can be included. Suitable are, e.g., 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 which carry a diethanolamino group, a methylamino group or a 2-methoxyethylamino group instead of the morpholino group. Furthermore, brighteners of the substituted 4,4'-distyryl- diphenyl type may be present, e.g., 4,4'-bis-(4-chloro-3-sulfostyryl)-diphenyl. Mixtures of brighteners can also be used. Brighteners of the 1 ,3-diaryl-2-pyrazoline type, e.g., 1-(p-sulfoamoylphenyl)-3-(p- chlorophenyl)-2-pyrazoline and compounds with a similar structure, are particularly suitable for polyamide fibers. The amount of optical brighteners or brightener mixtures in the composition is generally not more than 1 wt.%, preferably 0.05 to 0.5 wt.%.

The commonly used foam regulators that can be used in the compositions according to the invention include, e.g., 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. Further defoamers are kerosene hydrocarbons, in particular microparaffins and kerosene waxes whose melting point is above 40°C, saturated fatty acids or soaps with in particular 20 to 22 C atoms, e.g., sodium behenate, and alkali metal salts of phosphoric acid mono- and/or dialkyl esters in which the alkyl chains each have 12 to 22 C atoms. Among these, sodium monoalkyl phosphate and/or dialkyl phosphate with C16-18 alkyl groups is preferably used. The amount of foam regulators can preferably be 0.2 to 2 wt.%, particularly preferably not more than 1 wt.%.

To adjust the desired pH value, the compositions according to the invention may 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 sulphuric acid or alkali metal hydrogen sulphates, or bases, in particular ammonium or alkali metal hydroxides, preferably sodium hydroxide. Such pH regulators are contained in the compositions according to the invention preferably not more than 10 wt.%, in particular from 0.5 to 6 wt.%, particularly preferably from 0.3 to 2 wt.%.

In preferred embodiments, a composition according to the invention is characterized in that it comprises at least one solvent in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%, based on active substance and total weight of the composition.

In light of the invention, the addition of organic solvents is advantageous for the enzyme stability.

Preferred organic solvents are selected from the group consisting of monohydric or polyhydric alcohols, alkanolamines or glycol ethers. Preferably, the solvents are selected from the group consisting of ethanol, n- or i-propanol, butanol, glycol, propanediol, butanediol, glycerol, diglycol, pro pyldiglycol, butyldiglycol, hexylene glycol, ethylene glycol methyl ether, ethylene glycol ethyl ether, ethylene glycol propyl ether, ethylene glycol mono-n-butyl ether, diethylene glycol methyl ether, diethylene glycol methyl ether, propylene glycol methyl ether, propylene glycol ethyl ether, propylene glycol propyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, methoxy triglycol, ethoxy triglycol, butoxy triglycol, 1-butoxyethoxy-2-propanol, 3-methyl-3-methoxybutanol, propylene glycol t-butyl ether and mixtures of these solvents.

In more preferred embodiments, the solvent is selected from glycerol and 1 ,2-propandiol.

In even more preferred embodiments, a composition according to the invention is characterized in that it comprises, based on active substance and total weight of the composition, at least one solvent in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1.75%, more preferably 0.05 to 1.5 wt.%, most preferably 0.1 to 1 wt.%, wherein the solvent is selected from glycerol and 1 ,2-propandiol.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, an organic solvent, preferably selected from glycerol or 1 ,2 propanediol, in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%, at least one surfactant in an amount of 3 to 35 wt.%, preferably 5 to 30 wt.%, wherein the surfactant is selected from the group consisting of anionic surfactants, nonionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, a builder system, comprising at least one builder selected from the group consisting of polycarboxylic acids such as citric acid, adipic acid, succinic acid, glutaric acid, malic acid, tartaric acid, maleic acid, fumaric acid, sugar acids and carboxymethyl inulinulines or their salts, monomeric and polymeric amino polycarboxylic acids such as glycine diacetic acid, methyl glycine diacetic acid (MGDA), glutamic acid diacetic acid (GLDA), nitrile triacetic acid, iminodisuccinate such as ethylene diamine-N,N'-disuccinic acid and hydroxyamino disuccinates, ethylene diamine tetraacetic acid and polyaspartic acid or salts thereof, polyphosphonic acids such as amino tris-(methylene phosphonic acid), ethylene diamine tetrakis-(methylene phosphonic acid), lysine tetra-(methylene phosphonic acid), diethylene triamine penta-(methylene phosphonic acid) (DTPMP) and 1-hydroxyethane-1 ,1- diphosphonic acid (HEDP) or salts thereof, polymeric hydroxy compounds such as dextrin, and mixtures thereof, in an amount of 0.5 to 50 wt.%, preferably 0.5 to 20 wt.%, more preferably 0.5 to 10 wt.%, and at least one further enzyme in an amount of 1 x 10^-8 to 5 wt.%, preferably 0.0001 to 1 wt.%, more preferably 0.0005 to 0.5 wt.%, most preferably 0.001 to 0.1 wt.%, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases or mixtures thereof, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, an organic solvent, preferably selected from glycerol or 1 ,2 propanediol, in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of anionic surfactants, nonionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, a builder system, comprising at least one builder selected from the group consisting of polycarboxylic acids such as citric acid, adipic acid, succinic acid, glutaric acid, malic acid, tartaric acid, maleic acid, fumaric acid, sugar acids and carboxymethyl inulinulines or their salts, monomeric and polymeric amino polycarboxylic acids such as glycine diacetic acid, methyl glycine diacetic acid (MGDA), glutamic acid diacetic acid (GLDA), nitrile triacetic acid, imino disuccinate such as ethylene diamine-N,N'-disuccinic acid and hydroxyamino disuccinates, ethylene diamine tetraacetic acid and polyaspartic acid or salts thereof, polyphosphonic acids such as amino tris-(methylene phosphonic acid), ethylene diamine tetrakis-(methylene phosphonic acid), lysine tetra-(methylene phosphonic acid), diethylene triamine penta-(methylene phosphonic acid) (DTPMP) and 1-hydroxyethane-1 ,1- diphosphonic acid (HEDP) or salts thereof, polymeric hydroxy compounds such as dextrin, and mixtures thereof, in an amount of 0.5 to 50 wt.%, preferably 0.5 to 20 wt.%, more preferably 0.5 to 10 wt.%, and at least one further enzyme in an amount of 1 x 10^-8 to 5 wt.%, preferably 0.0001 to 1 wt.%, more preferably 0.0005 to 0.5 wt.%, most preferably 0.001 to 0.1 wt.%, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases or mixtures thereof, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, an organic solvent, selected from glycerol or 1 ,2 propanediol, in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and

LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, glycerol in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and

LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%,

1 ,2 propanediol in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

In preferred embodiments, a composition according to the invention comprises, in each cased based on active substance and total weight of the composition, at least one protease described herein in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, an organic solvent, preferably selected from glycerol or 1 ,2 propanediol, in an amount of 0.01 to 2 wt.%, preferably 0.02 to 1 .75%, more preferably 0.05 to 1 .5 wt.%, most preferably 0.1 to 1 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%,

LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, and at least one further enzyme in an amount of 1 x 10^-8 to 5 wt.%, preferably 0.0001 to 1 wt.%, more preferably 0.0005 to 0.5 wt.%, most preferably 0.001 to 0.1 wt.%, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases or mixtures thereof, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4- formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

A further subject of the invention relates to a method for cleaning textiles and/or hard surfaces, in particular dishes, which is characterized in that a composition according to the invention is used in at least one method step. In various embodiments, the method described is characterized in that the protease is used at a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

These include both manual and mechanical methods, whereby mechanical methods are preferred due to their more precise controllability, e.g., in terms of the quantities used and exposure times. Methods for cleaning textiles are generally characterized by the fact that various cleaning-active substances are applied to the items to be cleaned in several process steps and washed off after the exposure time, or that the items to be cleaned are otherwise treated with a washing composition or a solution or dilution of this composition.

All facts, subjects and embodiments described for the compositions according to the invention are also applicable to these objects of the invention. Therefore, at this point, explicit reference is made to the disclosure at the appropriate place with the indication that this disclosure also applies to the above-mentioned methods and uses according to the invention.

A further subject of the invention relates to the use of a washing and cleaning composition according to the invention and described herein for cleaning textiles and/or hard surfaces, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

A further subject of the invention relates to the use of a washing and cleaning composition according to the invention and described herein for improving the stability, in particular the storage stability, of enzymes, preferably proteases, in such washing and cleaning compositions, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

A further subject of the invention relates to the use of a washing and cleaning composition according to the invention and described herein for preventing and/or reducing the proteolysis of further enzymes contained in such washing and cleaning compositions by a protease contained in such washing and cleaning compositions, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

A further subject of the invention relates to the use of a washing and cleaning composition according to the invention and described herein for preventing and/or reducing the autoproteolysis of a protease contained in such washing and cleaning compositions, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

All facts, subjects and embodiments described for the compositions according to the invention are also applicable to these objects of the invention. Therefore, at this point, explicit reference is made to the disclosure at the appropriate place with the indication that this disclosure also applies to the above-mentioned methods and uses according to the invention.

EXAMPLES

Table 1 shows commercially available liquid detergents that was used for the following tests.

Table 1 : Washing compositions

Example 1 : Protease activity assay

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

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.

Sample preparation:

10 pL AAPF solution (70 mg/mL)

1000 pL Tris/HCI (0.1 M; pH 8.6 with 0.1% Brij 35)

10 pL diluted protease solution

Kinetics determined over 5 min at 25°C (410 nm)

Example 2: Storage stability of protease

The following protease variants were stored at 30°C and 40°C in 20 g of each formulas A, B and C shown in table 1 for 2, 4 and 8 weeks and their residual activity was determined by AAPF-pNA measurement. Protease 1 (P1): SEQ ID NO:1-S3T-V4I-R99E-V199I

Protease 2 (P2): SEQ ID NQ:1-S3T-V4l-R99E-S154D-V199l-Q200L-Y203W-A209K-L256E

The activity of the protease before storage (t = 0) was normalized to 100%. The protease activity after storage for 2 weeks (t = 2), 4 weeks (t = 4) and 8 weeks (t = 8) at 30°C and 40°C are indicated in relation to the start value. The results are shown in Table 2.

Table 2: Storage stability

BA = boric acid

From the results, it becomes apparent that protease P2 is more stable than protease P1 . Surprisingly, protease P2 is in formulas without boric acid even more stable than protease P1 in formulas with boric acid.

While protease P1 is more stable in formulas with higher boric acid amounts, protease P2 is surprisingly more stable in formulas without boric acid. This is insofar very surprising since boric acid is a well-known reversible protease inhibitor. The protease described herein thus presents a good option for boric acid free washing and cleaning compositions.

The effect is more prominent at higher storage temperatures and after longer storage duration.

In case of using detergent with higher solvent content (> 2 wt.% organic solvent), the effect of improved stability of protease P2 without boric acid in formula is even more prominent.

When using low solvent content, protease P2 is more stable without boric acid in formulas than protease P1 .

Example 3: Embodiments

Table 3: Liquid washing compositions

Table 4: Solid washing compositions

Claims

1 . Washing and cleaning composition, preferably liquid washing and cleaning composition, more preferably liquid laundry detergent, comprising, each based on the total weight of the washing and cleaning composition, at least one protease, in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein, wherein the protease has proteolytic activity and comprises an amino acid sequence which 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%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at at least two, preferably three, more preferably four, of the positions corresponding to positions 3, 4, 99 or 199, at least two, preferably three, more preferably four, amino acid substitutions selected from the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from N74D, N74E, N74Q, A136Q, R143L, R143W, R143Y, S154D, S154Q, S160G, Y161T, A163G, V171 L, A181 D, F183R, Q185R, Q200A, Q200L, Q200S, Q200T, Y203K, Y203V, Y203W, A209K, A209W, N212S, N212T, L256D, L256E, L256Q, preferably selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K und L256E, less than 0.2 wt.%, preferably less than 0.1 wt.%, more preferably less than 0.05 wt.%, most preferably 0 wt.% boric acid, at least one surfactant in an amount of 2 to 14.5 wt.%, preferably 3 to 13 wt.%, more preferably 5 to 12 wt.%, most preferably 7.5 to 10.5 wt.%, wherein the surfactant is selected from the group consisting of nonionic surfactants, anionic surfactants, cationic surfactants, zwitterionic surfactants, amphoteric surfactants and mixtures thereof, and optionally at least one further enzyme, wherein the enzyme is selected from the group consisting of amylases, lipases, mannanases, cellulases, pectate lyases and mixtures thereof, each in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%, based on active protein.

2. The washing and cleaning composition according to claim 1 , wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , (i) at the positions corresponding to positions 3, 4, 99 and 199, the amino acid substitutions S3T, V4I, R99E and V199I, and (ii) at at least one and increasingly preferably at least two, three, four or five, of the positions corresponding to positions 74, 136, 143, 154, 160, 161 , 163, 171 , 181 , 183, 185, 200, 203, 209, 212 or 256, at least one and increasingly preferably at least two, three, four or five, amino acid substitution(s) selected from S154D, S160G, A181 D, F183R, Q185R, Q200L, Y203W, A209K and L256E.

3. The washing and cleaning composition according to claim 1 or 2, wherein the protease has proteolytic activity and comprises an amino acid sequence which is at least 70% and increasingly preferably by at least 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% and less than 100% identical to the amino acid sequence specified in SEQ ID NO:1 over its entire length and which has, in each case based on the numbering according to SEQ ID NO:1 , one of the following amino acid substitution combinations:

(a) S3T-V4I-R99E-V199I-Q200L-Y203W;

(b) S3T-V4I-R99E-V199I-N212S;

(c) S3T-V4I-R99E-V199I-N74D;

(d) S3T-V4I-R99E-V199I-S154D-L256E;

(e) S3T-V4I-R99E-V199I-Q200L-Y203W-S154D-L256E;

(f) S3T-V4I-R99E-V199I-N74D-Q200L-Y203W;

(g) S3T-V4I-R99E-V199I-N74D-S154D-Q200L-Y203W-L256E;

(h) S3T-V4I-R99E-V199I-N74D-N212S;

(i) S3T-V4I-R99E-V199I-N74D-S154D-Y203W-L256E;

(j) S3T-V4I-R99E-V199I-N74D-Y203W;

(k) S3T-V4I-R99E-V199I-N74D-S154D-Q200L-L256E;

(l) S3T-V4I-R99E-V199I-N74D-Q200L;

(m) S3T-V4I-R99E-V199I-S154D-Q200L-Y203W;

(n) S3T-V4I-R99E-V199I-Q200L-Y203W-L256E;

(o) S3T-V4I-R99E-V199I-A136Q-R143W-Y161 T-Q200L;

(p) S3T-V4I-R99E-V199I-N74D-R143Y-A209W-N212S-L256E;

(q) S3T-V4I-R99E-V199I-A136Q-S154D-V171 L-Q200L;

(r) S3T-V4I-R99E-V199I-Q200L-Y203W-A209K-S154D-L256E;

(s) S3T-V4I-R99E-V199I-S154D-S160G-Q185R-Q200L-Y203W-L256E;

(t) S3T-V4I-R99E-V199I-S154D-A181 D-F183R-Q200L-Y203W-L256E;

(u) S3T-V4I-V199I-Q200L-Y203W;

(v) S3T-V4I-V199I-Q200L-Y203W-S154D-L256E;

(w) S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E;

(x) S3T-V4I-V199I-Q200I-Y203Y-A209W-S154D-L256E;

(y) S3T-V4I-V199I-Q200I-Y203W-A209W-S154D-L256E;

(z) S3T-V4I-V199I-Q200L-Y203W-A209W-S154D-L256E;

(aa) S3T-V4I-V199I-Q200I-Y203W-A209K-S154D-L256E;

(bb) S3T-V4I-V199I-Q200I-Y203Y-A209K-S154D-L256E;

(cc) S3T-V4I-V199I-Q200L-Y203W-A209K-S154D-L256E.

4. The washing and cleaning composition according to any one of the preceding claims, wherein the composition is free of boric acid.

5. The washing and cleaning composition according to any one of the preceding claims, wherein the composition is essentially free, preferably free, of any protease stabilizers selected from the group consisting of boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4-formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

6. The washing and cleaning composition according to any one of the preceding claims, comprising at least one protease according to any one of claims 1 to 3 in an amount of 0.0001 to 5 wt.%, preferably 0.001 to 2.5 wt.%, more preferably 0.005 to 1 wt.%, most preferably 0.01 to 0.5 wt.%,

FAEO in an amount of less than 6 wt.%, preferably 2.5 to 6 wt.%, more preferably 4 to 5.5 wt.%, most preferably 4.5 to 5.5 wt.%, and

LAS in an amount of less than 8.5 wt.%, preferably 1 to 8.5 wt.%, more preferably 2 to 7 wt.%, most preferably 2.5 to 5 wt.%, wherein the composition is free of any stabilizers selected from the group consisting of boric acids and salts thereof, boronic acids and salts thereof, in particular phenylboronic acid derivatives or 4-formylphenylboronic acid (4-FPBA), and peptide inhibitors, in particular peptide aldehydes.

7. The washing and cleaning composition according to any one of the preceding claims, wherein the composition is a liquid laundry detergent, wherein the composition has a pH value in 1 wt.% solution in deionized water at 20°C in a range from about 6 to about 11 , preferably from about 6.5 to about 10.5, more preferably from about 7 to about 10, most preferably from about 8 to about 9.

8. A method for cleaning textiles and/or hard surfaces, characterized in that a washing or cleaning composition according to any one of the preceding claims is used in at least one method step, the method preferably being carried out in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

9. Use of a washing or cleaning composition according to any one of claims 1 to 7 for cleaning textiles and/or hard surfaces, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.

10. Use of a washing or cleaning composition according to any one of claims 1 to 7 for improving the stability, in particular the storage stability, of enzymes, preferably proteases, in a washing and cleaning composition, preferably in a washing or cleaning composition according to any one of claims

1 to 7.

11 . Use of a washing or cleaning composition according to any one of claims 1 to 7 for preventing and/or reducing the proteolysis of further enzymes contained in the washing and cleaning composition by a protease contained in the washing and cleaning composition, wherein the washing and cleaning composition is preferably a washing and cleaning composition according to any one of claims 1 to 7.

12. Use of a washing or cleaning composition according to any one of claims 1 to 7 for preventing and/or reducing the autoproteolysis of a protease contained in the washing and cleaning composition, wherein the washing and cleaning composition is preferably a washing and cleaning composition according to any one of claims 1 to 7.

13. Use according to any one of claims 9 to 12, preferably in a temperature range from about 20°C to about 60°C, more preferably from about 20°C to about 40°C, most preferably from about 30°C to about 40°C.