DE102018004206A1 - Use of a polymer to increase the cleaning performance of a protease-containing detergent or cleaning agent - Google Patents

Abstract

The present invention relates to the use of at least one anionic polymer to increase the cleaning performance of a protease-containing washing or cleaning agent. Also part of the invention are the corresponding detergents and cleaners containing at least one protease and at least one anionic polymer, as well as the use of washing and cleaning agents and the corresponding washing and cleaning processes.

Description

The present invention relates to the use of at least one anionic polymer to increase the cleaning performance of a protease-containing washing or cleaning agent. Also part of the invention are the corresponding detergents and cleaners containing at least one protease and at least one anionic polymer, as well as the use of washing and cleaning agents and the corresponding washing and cleaning processes.

The use of enzymes in detergents and cleaners has been established in the art for decades. They serve to extend the range of services of the funds concerned according to their specific activities. These include in particular hydrolytic enzymes such as proteases, amylases, lipases and cellulases. The first three hydrolyze proteins, starches and fats and thus contribute directly to soil removal. Cellulases are used in particular because of their tissue effect. Another group of washing and cleaning agent enzymes are oxidative enzymes, in particular oxidases, which, if appropriate, in combination with other components, are preferably used to bleach soiling or to produce the bleaching agents in situ. In addition to these enzymes, which are subjected to constant optimization, further enzymes are constantly being made available for use in detergents and cleaners in order to be able to optimally address particular soiling, such as pectinases, β-glucanases, mannanases or other hemicellulases (glycosidases) Hydrolysis in particular of special vegetable polymers.

The enzymes which are the longest established and are contained in virtually all modern, powerful detergents and cleaners are proteases and, among these, in particular serine proteases, to which the subtilases according to the invention are also calculated. They serve the degradation of protein-containing stains on the cleaning material.

As detergency and detergent proteases various protease classes are established, for example metalloproteases. However, among the detergent and detergent proteases, subtilisin-type proteases (subtilases, subtilopeptidases, EC 3.4.21.62) occupy an outstanding position due to their favorable enzymatic properties such as stability or pH optimum. They are attributed to the serine proteases due to the catalytically active amino acids. They act as nonspecific endopeptidases, that is, they hydrolyze any acid amide linkages that are internal to peptides or proteins. Their pH optimum is usually in the clearly alkaline range. For an overview of this family, for example, the article "Subtilases: Subtilisin-like Proteases" by R. Siezen, pages 75-95 in "Subtilisin enzymes", edited by R. Bott and C. Betzel, New York, 1996 , Subtilases are naturally produced by microorganisms; Of these, in particular, the subtilisins formed and secreted by Bacillus species are to be mentioned as the most important group within the subtilases.

In general, only selected proteases are suitable for use in liquid surfactant-containing preparations. Many proteases do not show sufficient catalytic performance in such formulations. For the use of proteases in detergents, therefore, a high catalytic activity under conditions as they are during a wash cycle is particularly desirable.

From the international patent publications WO 2010/020475 and WO 2010/020476 It is known that certain substances can increase the performance of hydrolytic enzymes, in particular proteases. In these publications, inter alia, amino acids, polyamino acids and anionic / polyanionic polymers are mentioned as such substances.

Surprisingly, it has now been found that this performance-enhancing influence of special polymers on the performance of the protease in the washing or cleaning agent is dependent on the protease used and can be further increased by the special optimization of the enzyme for such a performance increase.

1. (a) an der Position, die der Position 99 entspricht, die Aminosäuresubstitution 99E; und
2. (b) an mindestens einer der Positionen, die den Positionen 76, 77, 188, 238, 242 und 245, jeweils bezogen auf die Nummerierung gemäß SEQ ID NO:1, entsprechen, mindestens eine weitere Aminosäuresubstitution aufweist.

The invention therefore in a first aspect, the use of at least one anionic or polyanionic polymer for increasing the cleaning performance of a protease-containing detergent or cleaning agent, characterized in that the protease is selected from a protease comprising an amino acid sequence which is at least 80% Sequence identity with the amino acid sequence given in SEQ ID NO: 1 over its entire length and in each case based on the numbering according to SEQ ID NO: 1

1. (a) at the position corresponding to position 99, the amino acid substitution 99E; and
2. (B) at least one of the positions corresponding to the positions 76, 77, 188, 238, 242 and 245, each based on the numbering of SEQ ID NO: 1, having at least one further amino acid substitution.

wobei

• a 0 oder 1 ist;
• X ausgewählt ist aus der Gruppe bestehend aus NH oder NR¹, wobei R¹ ausgewählt ist aus der Gruppe bestehend aus -CH₃ und -C₂H₅;
• R² mit jedem Vorkommen unabhängig ausgewählt ist aus der Gruppe bestehend aus -H und -CH₃;
• b eine ganze Zahl von 1 bis 5 ist;
• Y -(C=O)- ist;
• c 0 oder 1 ist;
• Z OH oder H ist; und
• d eine ganze Zahl von 10 bis 200 ist.

In various preferred embodiments of the invention, the polymer is selected from compounds of general structural formula (I):

in which

• a is 0 or 1;
• X is selected from the group consisting of NH or NR ¹ , wherein R ^{1 is} selected from the group consisting of -CH ₃ and -C ₂ H ₅ ;
• R ² with each occurrence is independently selected from the group consisting of -H and -CH ₃ ;
• b is an integer from 1 to 5;
• Y is - (C = O) -;
• c is 0 or 1;
• Z is OH or H; and
• d is an integer of 10 to 200.

• - Wasch- oder Reinigungsmittel enthaltend mindestens eine Protease und ein oben beschriebenes (poly)anionisches Polymer zur Steigerung der Reinigungsleistung des Wasch- oder Reinigungsmittels;
• - die Verwendung eines erfindungsgemäßen Wasch- oder Reinigungsmittels zum Waschen und/oder Reinigen von Textilien und/oder harten Oberflächen; sowie
• - Verfahren zur Reinigung von Textilien und/oder harten Oberflächen, die in mindestens einem Verfahrensschritt ein erfindungsgemäßes Wasch- oder Reinigungsmittel anwenden.

Further objects of the present invention relate to:

• - Washing or cleaning agent containing at least one protease and a (poly) anionic polymer described above to increase the cleaning performance of the detergent or cleaning agent;
• the use of a washing or cleaning agent according to the invention for washing and / or cleaning textiles and / or hard surfaces; such as
• - Process for the cleaning of textiles and / or hard surfaces, which use in one step at least one inventive detergent or cleaning agent.

wobei

• a 0 oder 1 ist;
• X ausgewählt ist aus der Gruppe bestehend aus NH oder NR¹, wobei R¹ ausgewählt ist aus der Gruppe bestehend aus -CH₃ und -C₂H₅;
• R² mit jedem Vorkommen unabhängig ausgewählt ist aus der Gruppe bestehend aus H und -CH₃;
• b eine ganze Zahl von 1 bis 5 ist;
• Y -(C=O) ist;
• c 0 oder 1 ist;
• Z OH oder H ist; und
• d eine ganze Zahl von 10 bis 200 ist;

wobei das Polymer ein Molekulargewicht (Mw) in einem Bereich von 1000 g/mol bis 20 000 g/mol, vorzugsweise von 2000 g/mol bis 10 000 g/mol, vorzugsweise 3000 g/mol bis 5000 g/mol aufweist und das Polymer in einer Menge von 0,5 bis 30 Gew.-%, vorzugsweise von 1 bis 20 Gew.-%, noch bevorzugter 2 bis 15 Gew.-% bezogen auf das Gesamtgewicht des Mittels in diesem enthalten ist.In various embodiments, the washing and cleaning agent according to the invention comprises at least one protease and at least one anionic or polyanionic polymer. In this case, the polymer can be selected from compounds of the general structural formula (I):

in which

• a is 0 or 1;
• X is selected from the group consisting of NH or NR ¹ , wherein R ^{1 is} selected from the group consisting of -CH ₃ and -C ₂ H ₅ ;
• R ² with each occurrence is independently selected from the group consisting of H and -CH ₃ ;
• b is an integer from 1 to 5;
• Y is - (C = O);
• c is 0 or 1;
• Z is OH or H; and
• d is an integer of 10 to 200;

wherein the polymer has a molecular weight (Mw) in a range from 1000 g / mol to 20 000 g / mol, preferably from 2000 g / mol to 10,000 g / mol, preferably 3000 g / mol to 5000 g / mol, and the polymer in an amount of 0.5 to 30 wt .-%, preferably from 1 to 20 wt .-%, more preferably 2 to 15 wt .-% based on the total weight of the composition in this is contained.

In various embodiments, the performance of the combination of protease and the polymer in the detergent may be further enhanced by the presence of calcium ions. Therefore, use in the presence of calcium ions as found in tap water is particularly preferred. It has been found that in the presence of calcium ions, preferably in concentrations of at least 1 mmol / L, more preferably at least 1.5 mmol / L, more preferably at least 2 mmol / L, macromolecular complexes of enzyme, polymer and calcium ions form. Therefore, use under conditions in which calcium ion concentrations of at least 1 mmol / L, more preferably at least 1.5 mmol / L, more preferably at least 2 mmol / L, are preferred under some circumstances.

These and other aspects, features, and advantages of the invention will become apparent to those skilled in the art from a study of the following detailed description and claims. Any feature of one aspect of the invention may be employed in any other aspect of the invention. Further, it is to be understood that the examples contained herein are intended to describe and illustrate the invention, but not to limit it, and in particular that the invention is not limited to these examples. All percentages are by weight unless otherwise specified, based on the corresponding composition or agent. Numeric ranges indicated in the format "from x to y" include the above values. If multiple preferred numeric ranges are specified in this format, it is understood that all ranges resulting from the combination of the various endpoints are also captured.

"At least one" as used herein refers to 1 or more, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or more.

According to the invention, a protease is to be understood as meaning all enzymes which are capable of hydrolyzing acid amide linkages of proteins. The proteases are also described in detail below.

The term "anionic or polyanionic polymer" as used herein refers to polymers that carry at least one and preferably several negative charges. It is preferably a polymer having at least one negatively charged monomer, preferably having a plurality of negatively charged monomers. Preferred are, for example, polymers of organic acids or their salts, in particular polyacrylates and / or poly-sugar acids and / or polyacrylate copolymers and / or poly-sugar copolymers. Of the aforementioned compounds, either the corresponding salts, in particular alkali metal salts, or else the free acids can be used. Further preferred compounds are polyacrylsulfonates or polycarboxylates and their salts, copolymers or salts of the copolymers. Preference is given, for example, to (poly) anionic polymers, in particular those which contain carboxylic acid monomers, in particular those which contain units derived from acrylic acid or methacrylic acid, very particularly polyacrylic or polymethacrylic acid or copolymers thereof and the corresponding salts. Likewise preferred are polymers based on aminocarboxylic acid monomers. Particular preference is given to using polyglutamic acid, including γ-D-polyglutamic acid, L-polyglutamic acid and DL-polyglutamic acid, polyaspartic acid, including β-D-polyaspartic acid and L-polyaspartic acid. An example of a particularly preferred polyaspartic acid is the compound available under the trade name Baypure DS 100 solid G (Lanxess company).

Further examples of particularly preferred substances are Acusol 587D (polyacrylic sulfonate, Rohm & Haas / Dow Chemical Company), Acusol 445N (polycarboxylate sodium salt, Rohm & Haas / Dow Chemical Company), Acusol 590 (polyacrylate copolymer; Rohm & Haas / Dow Chemical), Acusol 916 (polyacrylate sodium salt, Rohm & Haas / Dow Chemical), Sokalan CP42 (modified polycarboxylate sodium salt, BASF), Sokalan PA 30CL (polycarboxylate sodium salt, BASF), Dequest P 9000 (polymaleic acid, Thermphos), Alginic acid, poly-2 acrylamido-2-methyl-1-propane-sulfonic acid, poly-4-styrenesulfonic acid-co-maleic acid sodium salt, poly-acrylamido-coacrylic acid sodium salt, polymethacrylic acid sodium salt, poly-methylvinylether-old maleic acid, polyvinylsulfonic acid sodium salt, polyglutamic acid sodium salt, polyaspartic acid sodium salt.

In all embodiments described herein, the anionic / polyanionic polymers can be used in the form of the free acids or, preferably, in the form of their salts. In this case, in particular the alkali metal salts and especially the sodium salts are preferred.

• a 0 oder 1 ist;
• X NH ist;
• R² H ist;
• b 1 oder 2 ist;
• Y -(C=O)- ist;
• c 0 oder 1 ist; und
• d eine ganze Zahl von 30 bis 70 ist.

In some embodiments, the polymer as disclosed herein is characterized in that, in structural formula (I),

• a is 0 or 1;
• X is NH;
• R ^{2 is} H;
• b is 1 or 2;
• Y is - (C = O) -;
• c is 0 or 1; and
• d is an integer from 30 to 70.

In various embodiments, a and c are each 0, each R ² is independently H or methyl, and b is 1 to 4, preferably 1-3, more preferably 1-2, most preferably 1.

In various other embodiments, a and c are each 1. Preferably, R ^{2 is} H or methyl and b is 1 to 4, preferably 1-3, more preferably 1 or 2.

In various embodiments, when c is 0, Z is preferably H, when c is 1, Z at the - (C = O) terminus is preferably -OH.

In some embodiments, the polymer as disclosed herein is characterized in that the polymer is selected from the group consisting of polyacrylic acid, polyglutamic acid and polyaspartic acid.

In various embodiments, the invention also encompasses all stereoisomers, in particular enantiomers and diastereomers, tautomers and salts of the polymers described above. In the case of the salts, preference is given in particular to the alkali metal salts and in particular the sodium and potassium salts, especially the sodium salts.

The advantage of the polymers relevant to the invention over the prior art is that the cleaning performance of a protease-containing detergent or cleaning agent is enhanced by the polymer according to the invention without increasing the concentration of the protease. In this way, a less expensive washing or cleaning agent can be obtained, in which the concentration of the protease is kept low.

In addition, the polymers relevant to the invention have a good solubility in water, in particular because of the carboxyl groups present, so that they can easily be incorporated into corresponding agents and precipitation during storage is avoided.

1. (a) an der Position, die der Position 99 entspricht, die Aminosäuresubstitution 99E; und
2. (b) an mindestens einer der Positionen, die den Positionen 76, 77, 188, 238, 242 und 245, jeweils bezogen auf die Nummerierung gemäß SEQ ID NO:1, entsprechen, mindestens eine weitere Aminosäuresubstitution aufweist.

The proteases used according to the invention are those derived from the alkaline protease from Bacillus lentus having the amino acid sequence according to SEQ ID NO: 1. The protease is selected from a protease comprising an amino acid sequence which has at least 80% sequence identity with the amino acid sequence given in SEQ ID NO: 1 over its entire length and in each case based on the numbering according to SEQ ID NO: 1

1. (a) at the position corresponding to position 99, the amino acid substitution 99E; and
2. (B) at least one of the positions corresponding to the positions 76, 77, 188, 238, 242 and 245, each based on the numbering of SEQ ID NO: 1, having at least one further amino acid substitution.

In preferred embodiments of the protease used in the invention, the protease has at least one amino acid substitution according to (b) selected from the group consisting of 76H, 76K, 76W, 77Q, 77R, 188L, 238A, 238S, 238R, 238L, 238P, 238W, 242G , 242W, 245N, 245R and 245W, each based on the numbering according to SEQ ID NO: 1, is selected.

In various embodiments, the protease has only one amino acid substitution according to (b), which is selected from the group consisting of 76H, 188L, 238A, 238S, 238R, 238L, 238P, 238W, 245N and 245W, each based on the numbering according to SEQ ID NO: 1.

1. (i) In verschiedenen Ausführungsformen weist die Protease eine der folgenden Aminosäuresubstitutionsvarianten, jeweils bezogen auf die Nummerierung gemäß SEQ ID NO:1, auf:
2. (ii) R99E und A188L sowie optional zusätzliche eine von S76K, S76H, S76W, I77R, V238A, V238P, V238R, V238L, V238S, V238W, N242G, K245N und K245W;
3. (iii) R99E und eine von S76H, S76K und S76W sowie optional eine von I77R, V238A, V238P, V238L, V238S, V238W, N242G, K245N und K245W;
4. (iv) R99E und eine von V238A, V238S, V238R, V238L, V238W;
5. (v) R99E und V238R sowie eine von S76K, S76H, S76W, I77R, N242G, K245N und K245W.
6. (vi) R99E und N242G;
7. (vii) R99 und K245N oder K245W; und
8. (viii) R99 und I77R.

In various alternative embodiments, the protease has at least two amino acid substitutions according to (b) selected from 76W, 76K, 77R, 188L, 238A, 238R, 238P, 242G and 245N, preferably from 76W + 77R, 76K + 188L, 188L + 238P, 188L + 238R, 188L + 238A, 188L + 242G, 188I + 245N and 238R + 242G.

1. (i) In various embodiments, the protease has one of the following amino acid substitution variants, each based on the numbering according to SEQ ID NO: 1, on:
2. (ii) R99E and A188L and optionally additional ones of S76K, S76H, S76W, I77R, V238A, V238P, V238R, V238L, V238S, V238W, N242G, K245N and K245W;
3. (iii) R99E and one of S76H, S76K and S76W, and optionally one of I77R, V238A, V238P, V238L, V238S, V238W, N242G, K245N and K245W;
4. (iv) R99E and one of V238A, V238S, V238R, V238L, V238W;
5. (v) R99E and V238R and one of S76K, S76H, S76W, I77R, N242G, K245N and K245W.
6. (vi) R99E and N242G;
7. (vii) R99 and K245N or K245W; and
8. (viii) R99 and I77R.

In the aforementioned embodiments, the remainder of the amino acid sequence, i. the positions not mentioned as substituted, which correspond to SEQ ID NO: 1. Alternatively, however, the protease may also have further substitutions. For example, it is possible and contemplated by the invention that the proteases as described above contain one (or more) additional amino acid substitutions at one or more of the positions corresponding to positions 3, 4, 154, 199, and 256, based on the numbering SEQ ID NO: 1, preferably, one or more of the amino acid substitutions 3T, 41, 154D, 199I and 256E, more preferably 3T + 4I + 199I or 154D + 256E.

In one embodiment of the invention, the protease comprises an amino acid sequence which corresponds to the amino acid sequence given in SEQ ID NO: 1 over at least 80% of its total length, 81%, 82%, 83%, 84%, 85%, 86%, 87%. , 88%, 89%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5% or 98.8%, and in each case based on the numbering according to SEQ ID NO: 1 (a ) at the position corresponding to position 99, the amino acid substitution 99E; and (b) at least one of the positions corresponding to positions 76, 77, 188, 238, 242 and 245, each related to the numbering of SEQ ID NO: 1, has at least one more amino acid substitution. In the context of the present invention, the feature means that a protease has the indicated substitutions that it contains at least one of the corresponding amino acids at the corresponding positions, i. not all of the indicated positions are otherwise mutated or deleted, for example by fragmentation of the protease. Preferred substitutions are those selected from the group consisting of 76H, 76K, 76W, 77Q, 77R, 188L, 238A, 238S, 238R, 238L, 238P, 238W, 242G, 242W, 245N, 245R and 245W, each numbering according to SEQ ID NO: 1 are selected. Particularly preferred is the combination of 99E with one of 76H, 188L, 238A, 238S, 238R, 238L, 238P, 238W, 245N and 245W, each based on the numbering according to SEQ ID NO: 1. Also particularly preferred are combinations of 99E with at least 2 amino acid substitutions selected from 76W, 76K, 77R, 188L, 238A, 238R, 238P, 242G and 245N, preferably 76W + 77R, 76K + 188L, 188L + 238P, 188L + 238R, 188L + 238A, 188L + 242G, 188I + 245N and 238R + 242G.

The identity of nucleic acid or amino acid sequences is determined by a sequence comparison. This sequence comparison is based on the BLAST algorithm established in the prior art and commonly used (cf., for example, US Pat Altschul, SF, Gish, W., Miller, W., Myers, EW & Lipman, DJ (1990) "Basic local alignment search tool." J. Mol. Biol. 215: 403-410 , and Altschul, Stephan F, Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhang, Hheng Zhang, Webb Miller, and David J. Lipman (1997): "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs"; Nucleic Acids Res., 25, pp.3389-3402 ) and in principle occurs in that similar sequences of nucleotides or amino acids in the nucleic acid or amino acid sequences are assigned to each other. A tabular assignment of the respective positions is referred to as alignment. Another algorithm available in the prior art is the FASTA algorithm. Sequence comparisons (alignments), in particular multiple sequence comparisons, are created with computer programs. Frequently used, for example, the Clustal series (see, for example Chenna et al. (2003): Multiple sequence alignment with the Clustal series of programs. Nucleic Acid Research 31, 3497-3500 ), T-Coffee (cf., for example Notredame et al. (2000): T-Coffee: A novel method for multiple sequence alignments. J. Mol. Biol. 302, 205-217 ) or programs based on these programs or algorithms. Also possible are alignment comparisons (alignments) with the computer program Vector NTI® Suite 10.3 (Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, California, USA) with the default parameters whose AlignX module for sequence comparisons is based on ClustalW. Unless otherwise specified, the sequence identity given herein is determined by the BLAST algorithm.

Such a comparison also allows a statement about the similarity of the compared sequences to each other. It is usually given in percent identity, that is, the proportion of identical nucleotides or amino acid residues at the same or in an alignment corresponding positions. The broader concept of homology involves conserved amino acid substitutions in the consideration of amino acid sequences, that is, amino acids with similar chemical activity, as these usually perform similar chemical activities within the protein. Therefore, the similarity of the sequences compared may also be stated as percent homology or percent similarity. Identity and / or homology information can be made about whole polypeptides or genes or only over individual regions. Homologous or identical regions of different nucleic acid or amino acid sequences are therefore defined by matches in the sequences. Such areas often have identical functions. They can be small and comprise only a few nucleotides or amino acids. Often, such small regions exert essential functions for the overall activity of the protein. It may therefore be useful to relate sequence matches only to individual, possibly small areas. Unless otherwise indicated, identity or homology information in the present application, however, refers to the total length of the particular nucleic acid or amino acid sequence indicated.

In the context of the present invention, the indication that an amino acid position of a numerically designated position in SEQ ID NO: 1 corresponds to the corresponding position being assigned to the numerically designated position in SEQ ID NO: 1 in an alignment as defined above.

Functional fragments of the proteases described herein may also be employed. "Functional fragments" as used herein refers to enzymatically active polypeptides that are truncated N- and / or C-terminally by at least one, preferably two or more, amino acids as compared to the reference sequence. The activity of such fragments is at least 50%, preferably at least 60%, 70%, 80%, 90%, 95% or 100% of the activity of the reference enzyme.

1. (a) sie aus einer Protease wie oben beschrieben als Ausgangsmolekül erhältlich ist durch ein- oder mehrfache konservative Aminosäuresubstitution, wobei die Protease
   1. (i) an der Position, die der Position 99 entspricht, die Aminosäuresubstitution 99E; und
   2. (ii) an mindestens einer der Positionen, die den Positionen 76, 77, 188, 238, 242 oder 245, jeweils bezogen auf die Nummerierung gemäß SEQ ID NO:1, entsprechen, mindestens eine weitere Aminosäuresubstitution aufweist; und/oder
2. (b) sie aus einer Protease wie oben beschrieben als Ausgangsmolekül erhältlich ist durch Fragmentierung, Deletions-, Insertions- oder Substitutionsmutagenese und eine Aminosäuresequenz umfasst, die über eine Länge von mindestens 200, 210, 220, 230, 240, 250, 260, 261, 262, 263, 264, 265, 266, 267 oder 268 zusammenhängenden Aminosäuren mit dem Ausgangsmolekül übereinstimmt, wobei die Protease
   1. (i) an der Position, die der Position 99 entspricht, die Aminosäuresubstitution 99E; und
   2. (ii) an mindestens einer der Positionen, die den Positionen 76, 77, 188, 238, 242 oder 245, jeweils bezogen auf die Nummerierung gemäß SEQ ID NO:1, entsprechen, mindestens eine weitere Aminosäuresubstitution aufweist.

In various embodiments, the protease is characterized in that

1. (a) it is obtainable from a protease as described above as the starting molecule by single or multiple conservative amino acid substitution, wherein the protease
   1. (i) at the position corresponding to position 99, the amino acid substitution 99E; and
   2. (ii) at least one of the positions corresponding to positions 76, 77, 188, 238, 242 or 245, each based on the numbering according to SEQ ID NO: 1, has at least one further amino acid substitution; and or
2. (b) it is obtainable from a protease as described above as the starting molecule by fragmentation, deletion, insertion or substitution mutagenesis and comprises an amino acid sequence of at least 200, 210, 220, 230, 240, 250, 260, 261 , 262, 263, 264, 265, 266, 267 or 268 contiguous amino acids matches the parent molecule, the protease
   1. (i) at the position corresponding to position 99, the amino acid substitution 99E; and
   2. (ii) at least one of the positions corresponding to the positions 76, 77, 188, 238, 242 or 245, each based on the numbering according to SEQ ID NO: 1, has at least one further amino acid substitution.

In further embodiments of the invention, the protease obtainable by such a method is still at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90 , 5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%, 94.5%, 95%, 95.5%, 96%, 96.5 %, 97%, 97.5%, 98%, 98.5%, or 98.8% identical to the amino acid sequence given in SEQ ID NO: 1 over their entire length.

In detergents or cleaners according to the invention, which in one embodiment are predominantly solid and, in another embodiment, are predominantly liquid, pasty or gel, the enzyme, ie the protease, is present in an amount of 1 × 10 ^-8 to 5 wt % by weight of active protein, more preferably in an amount of from 1 × 10 ^-7 to 3% by weight, from 0.00001 to 1% by weight, from 0.00005 to 0.5% by weight, from 0.0001 to 0.1 wt .-% and particularly preferably from 0.0001 to 0.05 wt .-%, based on the active protein and the total weight of the washing or cleaning agent, are contained in this.

In various embodiments, the protease, and optionally also the polymer, may be pre-formulated in an enzyme composition. The enzyme protein typically accounts for only a fraction of the total weight of common enzyme preparations. Preferably used protease preparations contain from 0.1 to 40 wt .-%, preferably from 0.2 to 30 wt .-%, particularly preferably from 0.4 to 20 wt .-% and in particular from 0.8 to 10 wt .-% of the protease, in each case based on the active protein and the total weight of the enzyme preparation. In such compositions, the polymer may be contained in an amount of from 0.05 to 35% by weight, preferably from 0.05 to 10% by weight, based on the total weight. This enzyme composition, which is likewise a constituent of the present invention, can then be used in detergents or cleaners according to the invention in amounts which lead to the above-mentioned final concentrations in the washing or cleaning agent.

The protein concentration can be determined by known methods, for example the BCA method (bicinchoninic acid, 2,2'-biquinolyl-4,4'-dicarboxylic acid) or the biuret method. The determination of the active protein concentration takes place in this respect via a titration of the active sites using a suitable irreversible inhibitor (for proteases, for example, phenylmethylsulfonyl fluoride (PMSF)) and determination of the residual activity (see. Bender, M., et al., J. Am. Chem. Soc. 88, 24 (1966), p. 5890-5913 ).

In addition to the polymer according to the general formula given above, an inventive composition or an enzyme preparation used according to the invention at least one stabilizer, in particular a polyol such as glycerol or 1,2-ethylene glycol, and / or an antioxidant, but also inhibitors such as boric acid or Derivatives thereof or peptide inhibitors.

In the agents described herein, the enzymes to be used, for example proteases, may also be formulated together with adjuncts, for example from the fermentation. In liquid formulations, the enzymes are preferably used as enzyme liquid formulation (s).

As a rule, the proteases are not provided in the form of the pure protein but, as already mentioned above, in the form of stabilized, storage and transportable preparations. Such prefabricated preparations include, for example, the solid preparations obtained by granulation, extrusion or lyophilization or, especially in the case of liquid or gel-form detergents, solutions of the enzymes, advantageously as concentrated as possible, low in water and / or added with stabilizers or further auxiliaries.

Alternatively, the enzymes may be encapsulated for both the solid and liquid dosage forms, for example by spray-drying or extruding the enzyme solution together with a preferably natural polymer or in the form of capsules, for example those in which the enzymes are entrapped as in a solidified gel or in those of the core-shell type, in which an enzyme-containing core is coated with a water, air and / or chemical impermeable protective layer. In deposited layers, further active ingredients, for example stabilizers, emulsifiers, pigments, bleaches or dyes, may additionally be applied. Such capsules are applied by methods known per se, for example by shaking or rolling granulation or in fluid bed processes. Advantageously, such granules, for example by applying polymeric film-forming agent, low in dust and storage stable due to the coating.

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

Agents according to the invention may contain, in addition to the protease, one or more further enzymes, in particular from the following group: further proteases, amylases, hemicellulases, cellulases, lipases and oxidoreductases.

The amylase (s) is preferably an α-amylase. The hemicellulase is preferably a β-glucanase, a pectinase, a pullulanase and / or a mannanase. The cellulase is preferably a cellulase mixture or a one-component cellulase, preferably or predominantly an endoglucanase and / or a cellobiohydrolase. The oxidoreductase is preferably an oxidase, in particular a choline oxidase, or a perhydrolase.

The agents described herein include all conceivable types of detergents or cleaners, both concentrates and undiluted agents, for use on a commercial scale, in the washing machine, or in hand washing. These include detergents for textiles, carpets, or natural fibers, for which the term detergent is used. These include, for example, dishwashing detergents for dishwashers or manual dishwashing detergents or cleaners for hard or solid surfaces such as metal, glass, porcelain, ceramics, tiles, stone, painted surfaces, plastics, wood or leather, for which the term detergent is used, ie next to Manual and automatic dishwashing detergents, for example abrasives, glass cleaners, toilet scavengers, etc. The washing and cleaning agents in the invention also include washing aids, which are added to the actual detergent in the manual or machine textile laundry to achieve a further effect. Furthermore, laundry detergents and cleaners in the context of the invention also include textile pre-treatment and post-treatment agents, ie those agents with which the laundry item is brought into contact before the actual laundry, for example to dissolve stubborn soiling, and also agents which are in one of the actual Textile laundry downstream step to give the laundry further desirable properties such as comfortable grip, crease resistance or low static charge. Among the latter, i.a. calculated the fabric softener.

Embodiments of the present invention include all solid, powdered, liquid, gelatinous or pasty administration forms of agents described herein, which if appropriate can also consist of several phases and can be present in compressed or uncompressed form. The agent can 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. The solid dosage forms of the composition also include extrudates, granules, tablets or pouches. Alternatively, the agent can also be liquid, gelatinous or pasty, for example in the form of a non-aqueous liquid washing or dishwashing detergent or a non-aqueous paste or in the form of an aqueous liquid washing or dishwashing detergent or a water-containing paste.

Furthermore, the agent may be present as a one-component system. Such funds consist of one phase. Alternatively, an agent can also consist of several phases. Such an agent is therefore divided into several components.

The detergents or cleaners described herein, which may be in the form of powdered solids, in densified particulate form, as homogeneous solutions or suspensions, may further additionally contain all known ingredients customary in such compositions, preferably at least one further ingredient being present in the composition. In particular, the agents described herein may contain surfactants, builders, bleaches or bleach activators. Furthermore, they may contain water-miscible organic solvents, sequestering agents, electrolytes, pH regulators and / or further auxiliaries, such as optical brighteners, grayness inhibitors, foam regulators, as well as dyes and fragrances, and combinations thereof.

Advantageous ingredients of the agents described herein are disclosed in the international patent application WO2009 / 121725 starting there on page 5, penultimate paragraph, and ending on page 13 after the second paragraph. This disclosure is incorporated herein by reference and the disclosure is incorporated herein by reference.

Another object is the use of a washing or cleaning agent described herein for washing textiles or cleaning solid surfaces, especially in washing and cleaning processes.

A further object is a process for the cleaning of textiles or hard or solid surfaces, which is characterized in that in at least one process step, an agent described herein is used or that in at least one process step, a protease of the invention is catalytically enhanced by the polymer according to the invention, in particular such that the protease is used in an amount of 40 μg to 4 g, preferably from 50 μg to 3 g, particularly preferably from 100 μg to 2 g and very particularly preferably from 200 μg to 1 g, together with the polymer.

In various embodiments, such methods are characterized in that the protease is used at a temperature of 0-100 ° C, preferably 0-60 ° C, more preferably 20-45 ° C, and most preferably at 40 ° C. In addition, such processes are characterized in that the agents containing the protease are used in combination with the polymer in the form of a calcium-containing wash liquor.

These include both manual and mechanical processes, with mechanical processes being preferred. Processes for cleaning textiles are generally distinguished by the fact that various cleaning-active substances are applied to the items to be cleaned and washed off after the action time, or that the items to be cleaned are otherwise treated with a detergent or a solution or dilution of this agent. The same applies to processes for the purification of all materials other than textiles, especially hard or solid surfaces. All conceivable washing or cleaning methods can be enriched in at least one of the method steps by the use of a washing or cleaning agent described herein and then represent embodiments of the present invention.

All aspects, objects, and embodiments described for means described herein are also applicable to the aforementioned methods and uses. Therefore, reference is made at this point expressly to the disclosure in the appropriate place with the statement that this disclosure also applies to the above described methods and uses.

Examples

Example 1: Activity determination of the protease

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

The measurement was carried out at a temperature of 25 ° C, at pH 8.6 and a wavelength of 410 nm. The measuring time was 5 min at a measuring interval of 20 to 60 seconds.

Measurement Approach:

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

Example 2: Mini wash test

The miniwash test was performed on the shake flask supernatant of the proteases produced.

Conditions: 40 ° C, 16 ° dH water, 1h

Detergent Concentration: 4.52 g / L (Composition According to Table 3) Enzyme Concentration: The supernatants are washed in the same activity compared to a benchmark with the protease according to SEQ ID NO: 1, with a market-standard concentration for proteases in detergent

Polymer concentration: 0.11 g / mL polyacrylic acid or 0.11 g / mL polyglutamic acid in the detergent

stains:

• 10N (whole egg, pigment)
• C-03 (chocolate milk and soot)
• C-05 (blood, milk, ink)

• • Ausgestanzte Gewebe (Durchmesser = 10 mm) in Mikrotiterplatte vorlegen, Waschlauge auf 40°C vortemperieren, Endkonzentration 4,52 g/L in 16°dH Wasser
• • Lauge, Enzym und Polymer auf die Anschmutzung geben, für 1 h bei 40°C und 600 rpm inkubieren,
• • anschließend die Anschmutzung mehrmals mit klarem Wasser spülen, trocken lassen und mit einem Farbmessgerät die Helligkeit bestimmen.

Execution:

• • Place excised tissue (diameter = 10 mm) in a microtiter plate, preheat the wash liquor to 40 ° C, final concentration 4.52 g / L in 16 ° dH water
• • place lye, enzyme and polymer on the soil, incubate for 1 h at 40 ° C and 600 rpm,
• • then rinse the soiling several times with clean water, leave to dry and determine the brightness with a colorimeter.

The lighter the tissue, the better the cleaning performance. The L-value = brightness is measured, the higher the brighter. It is washed with a common liquid detergent without enzymes.

Tabelle 1: Ergebnisse des Miniwaschtests Waschmittel mit Proteasevarianten und Polyacrylsäure (PAA; Acusol 445N) 10 N C 03 C05 0,05% PAA P_PAA 4,5 -0,4 2,3 R99E P_Protease 4,1 5,0 14,7 P_protease+PAA 11,9 6,8 21,6 P_Netto-Boost 3,3 2,2 4,6 R99E+S76H P_Protease 1,9 5,6 16,0 P_protease+PAA 12,0 7,9 20,4 P_Netto-Boost 5,7 2,7 2,0 R99E+A188L P_Protease 4,9 7,3 17,7 P_protease+PAA 13,3 10,3 24,3 P_Netto-Boost 3,9 3,4 4,3 R99E+V238A P_Protease 5,0 7,4 18,3 P_protease+PAA 13,1 10,3 23,3 P_Netto-Boost 3,6 3,3 2,6 R99E+V238S P_Protease 6,4 8,4 18,8 P_protease+PAA 12,7 9,8 24,4 P_Netto-Boost 1,8 1,8 3,2 R99E+V238R P_Protease 5,7 5,3 16,3 P_protease+PAA 12,8 7,4 21,4 P_Netto-Boost 2,5 2,6 2,8 R99E+V238L P_Protease 3,3 6,6 18,6 P_protease+PAA 11,0 8,7 22,5 P_Netto-Boost 3,2 2,5 1,6 R99E+V238P P_Protease 6,2 6,9 18,6 P_protease+PAA 13,7 9,9 24,3 P_Netto-Boost 3,1 3,4 3,4 R99E+V238W P_Protease 6,3 7,5 18,8 P_protease+PAA 12,8 9,1 24,8 P_Netto-Boost 2,0 2,1 3,6 R99E+K245N P_Protease 2,9 7,7 17,2 P_protease+PAA 11,4 10,5 22,8 P_Netto-Boost 4,0 3,2 3,2 R99E+K245W P_Protease 4,2 7,0 23,1 P_protease+PAA 13,6 8,9 17,8 P_Netto-Boost 4,9 2,3 -7,6 Tabelle 2: Ergebnisse des Miniwaschtests für das Waschmittel mit Proteasevarianten und Polyglutaminsäure (PGA) 10 N C 03 C05 0,05% PGA P_PGA 1,6 5,0 0,3 R99E P_Protease 4,1 5,0 14,7 P_protease+PAA 6,8 6,3 16,3 P_Netto-Boost 1,1 n.d. 1,3 R99E+S76H P_Protease 1,9 13,4 16,0 P_protease+PGA 6,8 17,1 18,7 P_Netto-Boost 3,4 2,4 2,4 R99E+A188L P_Protease 4,9 14,9 17,7 P_protease+PGA 8,1 18,8 20,0 P_Netto-Boost 1,6 1,1 2,0 R99E+V238A P_Protease 5,0 14,3 18,3 P_protease+PGA 10,0 18,3 19,7 P_Netto-Boost 3,5 0,7 1,1 R99E+V238S P_Protease 6,4 14,5 18,8 P_protease+PGA 8,5 19,3 21,5 P_Netto-Boost 0,5 1,3 2,4 R99E+V238R P_Protease 5,7 14,0 16,3 P_protease+PGA 8,5 16,2 18,5 P_Netto-Boost 1,2 1,7 1,9 R99E+V238L P_Protease 3,3 14,0 18,6 _{Pprotease+PGA} 7,4 17,1 17,3 P_Netto-Boost 2,6 0,4 n.d R99E+V238P P_Protease 6,2 15,0 18,6 P_protease+PGA 9,3 17,4 19,5 P_Netto-Boost 1,5 0,6 0,7 R99E+K245N P_Protease 2,9 12,3 17,2 P_protease+PGA 6,9 18,4 19,5 P_Netto-Boost 2,5 1,3 2,0 R99E+K245W P_Protease 4,2 12,7 23,1 P _protease+PGA 8,2 17,4 19,6 P_Netto-Boost 2,5 1,0 n.d Tabelle 3: Ergebnisse des Miniwaschtests (L-Wert (Variante) - L-Wert (R99E)) Variante 10 N C 03 C05 R99E+K245N+A188L 1,4 5,1 1,1 R99E+V238P+A188L 1,8 4,1 7,0 R99E+V238R+A188L 3,1 3,0 2,4 R99E+S76H+A188L 0,1 4,0 7,7 R99E+V238A+A188L 0,1 3,7 -1,9 R99E+V238R+N242G 1,8 4,4 7,6 R99E+S76W+I77R 0,1 3,7 4,0 R99E+A188L+N242G 6,2 6,6 4,7 Calculation of the net performance increase: $${\text{P}}_{\text{net}-\text{performance increase}}=\left({\text{P}}_{\text{protease}+\text{PAA}}-{\text{P}}_{\text{bright}}\right)-\left({\text{P}}_{\text{PAA}}-{\text{P}}_{\text{bright}}\right)-\left({\text{P}}_{\text{protease}}-{\text{P}}_{\text{bright}}\right)$$

Table 1: Results of the mini-wash test Detergent with protease variants and polyacrylic acid (PAA; Acusol 445N) 10 N C 03 C05 0.05% PAA _PAA 4.5 -0.4 2.3 R99E P _protease 4.1 5.0 14.7 P _{protease + PAA} 11.9 6.8 21.6 P _{net boost} 3.3 2.2 4.6 R99E + S76H P _protease 1.9 5.6 16.0 P _{protease + PAA} 12.0 7.9 20.4 P _{net boost} 5.7 2.7 2.0 R99E + A188L P _protease 4.9 7.3 17.7 P _{protease + PAA} 13.3 10.3 24.3 P _{net boost} 3.9 3.4 4.3 R99E + V238A P _protease 5.0 7.4 18.3 P _{protease + PAA} 13.1 10.3 23.3 P _{net boost} 3.6 3.3 2.6 R99E + V238S P _protease 6.4 8.4 18.8 P _{protease + PAA} 12.7 9.8 24.4 P _{net boost} 1.8 1.8 3.2 R99E + V238R P _protease 5.7 5.3 16.3 P _{protease + PAA} 12.8 7.4 21.4 P _{net boost} 2.5 2.6 2.8 R99E + V238L P _protease 3.3 6.6 18.6 P _{protease + PAA} 11.0 8.7 22.5 P _{net boost} 3.2 2.5 1.6 R99E + V238P P _protease 6.2 6.9 18.6 P _{protease + PAA} 13.7 9.9 24.3 P _{net boost} 3.1 3.4 3.4 R99E + V238W P _protease 6.3 7.5 18.8 P _{protease + PAA} 12.8 9.1 24.8 P _{net boost} 2.0 2.1 3.6 R99E + K245N P _protease 2.9 7.7 17.2 P _{protease + PAA} 11.4 10.5 22.8 P _{net boost} 4.0 3.2 3.2 R99E + K245W P _protease 4.2 7.0 23.1 P _{protease + PAA} 13.6 8.9 17.8 P _{net boost} 4.9 2.3 -7.6 Table 2: Results of the mini-washing test for the detergent with protease variants and polyglutamic acid (PGA) 10 N C 03 C05 0.05% PGA P _PGA 1.6 5.0 0.3 R99E P _protease 4.1 5.0 14.7 P _{protease + PAA} 6.8 6.3 16.3 P _{net boost} 1.1 nd 1.3 R99E + S76H P _protease 1.9 13.4 16.0 P _{protease + PGA} 6.8 17.1 18.7 P _{net boost} 3.4 2.4 2.4 R99E + A188L P _protease 4.9 14.9 17.7 P _{protease + PGA} 8.1 18.8 20.0 P _{net boost} 1.6 1.1 2.0 R99E + V238A P _protease 5.0 14.3 18.3 P _{protease + PGA} 10.0 18.3 19.7 P _{net boost} 3.5 0.7 1.1 R99E + V238S P _protease 6.4 14.5 18.8 P _{protease + PGA} 8.5 19.3 21.5 P _{net boost} 0.5 1.3 2.4 R99E + V238R P _protease 5.7 14.0 16.3 P _{protease + PGA} 8.5 16.2 18.5 P _{net boost} 1.2 1.7 1.9 R99E + V238L P _protease 3.3 14.0 18.6 _{Pprotease + PGA} 7.4 17.1 17.3 P _{net boost} 2.6 0.4 nd R99E + V238P P _protease 6.2 15.0 18.6 P _{protease + PGA} 9.3 17.4 19.5 P _{net boost} 1.5 0.6 0.7 R99E + K245N P _protease 2.9 12.3 17.2 P _{protease + PGA} 6.9 18.4 19.5 P _{net boost} 2.5 1.3 2.0 R99E + K245W P _protease 4.2 12.7 23.1 P _{protease + PGA} 8.2 17.4 19.6 P _{net boost} 2.5 1.0 nd Table 3: Results of the mini-wash test (L value (variant) - L value (R99E)) variant 10 N C 03 C05 R99E + K245N + A188L 1.4 5.1 1.1 R99E + V238P + A188L 1.8 4.1 7.0 R99E + V238R + A188L 3.1 3.0 2.4 R99E + S76H + A188L 0.1 4.0 7.7 R99E + V238A + A188L 0.1 3.7 -1.9 R99E + + V238R N242G 1.8 4.4 7.6 R99E + + S76W I77R 0.1 3.7 4.0 R99E + A188L + N242G 6.2 6.6 4.7

From the results of the wash tests, it is clear that, for all the proteases listed, the combination of protease and polymer in the detergent results in a better wash performance than if the wash performance of the polymer and the protease alone were summed up. This shows a clear synergistic effect between the protease variants and polyacrylic acid or polyglutamic acid. This causes an improved washing performance on protein-containing stains. Table 3: Detergent matrix used Chemical name Wt .-% active substance in the raw material Wt .-% active ingredient in the formulation Water demin. 100 rest alkyl benzene sulfonic acid 96 4.40 Other anionic surfactants 70 5.60 C12-C18 fatty acids Na salt 30 2.40 Nonionic surfactants 100 4.40 phosphonates 40 0.20 citric acid 100 1.40 NaOH 50 0.95 defoamers tq 0.01 glycerin 100 2.00 preservative 100 0.08 ethanol 93 1,00

The matrix used contained no optical brighteners, perfumes, dyes or enzymes.

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• WO 2010/020475 [0006]
• WO 2010/020476 [0006]
• WO 2009/121725 [0053]

Cited non-patent literature

• "Subtilases: Subtilisin-like Proteases" by R. Siezen, pages 75-95 in "Subtilisin enzymes", edited by R. Bott and C. Betzel, New York, 1996. [0004]
• Altschul, S.F., Gish, W., Miller, W., Myers, E.W. & Lipman, D.J. (1990) "Basic local alignment search tool." J. Mol. Biol. 215: 403-410 [0033]
• Altschul, Stephan F., Thomas L. Madden, Alejandro A. Schaffer, Jinghui Zhang, Hheng Zhang, Webb Miller, and David J. Lipman (1997): "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs "; Nucleic Acids Res., 25, p.3389-3402 [0033]
• Chenna et al. (2003): Multiple sequence alignment with the Clustal series of programs. Nucleic Acid Research 31, 3497-3500 [0033]
• Notredame et al. (2000): T-Coffee: A novel method for multiple sequence alignments. Biol. 302, 205-217 [0033]
• Bender, M., et al., J. Am. Chem. Soc. 88, 24 (1966), pp. 5890-5913 [0041]

Claims (11)

Use of at least one anionic or polyanionic polymer to increase the cleaning performance of a protease-containing detergent or cleaning agent, characterized in that the protease is selected from a protease comprising an amino acid sequence having at least 80% sequence identity with that given in SEQ ID NO: 1 Has amino acid sequence over the entire length thereof and in each case based on the numbering according to SEQ ID NO: 1 a) at the position corresponding to the position 99, the amino acid substitution 99E; and b) at least one of the positions corresponding to the positions 238, 188, 76, 77, 242 and 245, each based on the numbering according to SEQ ID NO: 1, has at least one further amino acid substitution. Use after Claim 1 characterized in that the polymer is selected from compounds of general structural formula (I):

where a is 0 or 1; X is selected from the group consisting of NH or NR ¹ , wherein R ^{1 is} selected from the group consisting of -CH ₃ and -C ₂ H ₅ ; R ² with each occurrence is independently selected from the group consisting of -H and -CH ₃ ; b is an integer from 1 to 5; Y is - (C = O) -; c is 0 or 1; Z is OH or H; and d is an integer of 10 to 200. Use after Claim 1 or 2 , characterized in that the polymer in an amount of 0.5 to 30 wt .-%, preferably from 1 to 20 wt .-%, more preferably 2 to 15 wt .-% based on the total weight of the detergent or cleaning agent in this is included. Use according to one of Claims 1 to 3 characterized in that in formula (I), (A1) a is 0 or 1; X is NH; R ^{2 is} -H; b is 1 or 2; Y is - (C = O) -; c is 0 or 1; and d is an integer of 30 to 70; or (A2) a and c are each 0; each R ^{2 is} independently H or methyl; and b is 1 to 4, preferably 1-3, more preferably 1-2, most preferably 1; or (A3) a and c are each 1; each R ^{2 is} independently H or methyl; and b is 1 to 4, preferably 1-3, more preferably 1 or 2. Use according to one of Claims 1 to 4 , characterized in that the polymer is selected from the group consisting of polyacrylic acid, polyglutamic acid and polyaspartic acid. Use according to one of Claims 1 to 5 characterized in that the one or more amino acid substitutions of the protease according to b) are selected from the group 238P, 188L, 76H, 76K, 76W, 77Q, 77R, 238A, 238S, 238R, 238L, 238W, 242G, 242W, 245N, 245R and 245W, each based on the numbering according to SEQ ID NO: 1. Use after Claim 6 characterized in that (A) the protease has only one amino acid substitution according to (b), which is selected from the group consisting of 76H, 188L, 238A, 238S, 238R, 238L, 238P, 238W, 245N and 245W, respectively to the numbering according to SEQ ID NO: 1; or (B) the protease has at least two amino acid substitutions according to (b) selected from 76W, 76K, 77R, 188L, 238A, 238R, 238P, 242G and 245N, preferably from 76W + 77R, 76K + 188L, 188L + 238P, 188L + 238R, 188L + 238A, 188L + 242G, 188I + 245N and 238R + 242G; or (C) the protease has one of the following amino acid substitution variants, in each case based on the numbering according to SEQ ID NO: 1, (i) R99E and A188L and optionally additional ones of S76K, S76H, S76W, I77R, V238A, V238P, V238R, V238L, V238S, V238W, N242G, K245N and K245W; (ii) R99E and one of S76H, S76K and S76W, and optionally one of I77R, V238A, V238P, V238L, V238S, V238W, N242G, K245N and K245W; (iii) R99E and one of V238A, V238S, V238R, V238L, V238W; (iv) R99E and V238R; and one of S76K, S76H, S76W, I77R, N242G, K245N and K245W. (v) R99E and N242G; (vi) R99 and K245N or K245W; and (vii) R99 and I77R. Use according to one of Claims 1 to 7 , characterized in that the protease-containing washing or cleaning agent comprises at least one protease; wherein the at least one protease in an amount of 1 × 10 ^-8 to 5 wt .-% based on active protein, more preferably in an amount of 1 × 10 ^-7 to 3 wt .-%, from 0.00001 to 1 wt .-%, from 0.00005 to 0.5 wt .-%, from 0.0001 to 0.1 wt .-% and particularly preferably from 0.0001 to 0.05 wt .-%, based on active protein and the Total weight of the washing or cleaning agent, are included in this. Detergent or cleaning agent containing (a) at least one protease comprising an amino acid sequence which has at least 80% sequence identity with the amino acid sequence given in SEQ ID NO: 1 over its entire length and in each case based on the numbering according to SEQ ID NO: 1 (i) the position corresponding to the position 99, the amino acid substitution 99E; and (ii) at least one of the positions corresponding to positions 76, 77, 188, 238, 242 and 245, each based on the numbering of SEQ ID NO: 1, has at least one further amino acid substitution; and (b) at least one polymer, wherein the polymer is selected from compounds of general structural formula (I):

where - a is 0 or 1; X is selected from the group consisting of NH or NR1, wherein R ^{1 is} selected from the group consisting of -CH ₃ and -C ₂ H ₅ ; R ^{2 is} selected from the group consisting of -H and -CH ₃ ; b is an integer from 1 to 5; - Y - (C = O) is; - c is 0 or 1; - Z is OH or H; and d is an integer of from 10 to 200, the polymer having a molecular weight Mw in a range of from 1,000 to 20,000 g / mol, preferably from 2,000 to 10,000 g / mol; wherein the polymer is contained therein in an amount of 0.5 to 30% by weight, preferably 1 to 20% by weight, more preferably 2 to 15% by weight based on the total weight of the agent. Use of a washing or cleaning agent after Claim 9 for washing textiles or cleaning solid surfaces. Process for the cleaning of textiles or hard surfaces, characterized in that in at least one process step, a washing or cleaning agent after Claim 9 is applied.