Cleaning agent containing amylases

Abstract

A cleaning agent for hard surfaces and methods for cleaning are provided herein. In one embodiment, the cleaning agent includes at least one first amylase, wherein the first amylase is an α-amylase from Bacillus sp. No. 707 or a functional fragment or a variant thereof. The cleaning agent further includes at least one second amylase, wherein the second amylase is an AA560 α-amylase from Bacillus sp. or a functional fragment or a variant thereof. In another embodiment, the method includes dispensing the cleaning agent into the interior of an automatic dishwasher while a dishwashing program is being executed, before the main washing cycle begins, or in the course of the main washing cycle.

1 . A cleaning agent for hard surfaces, comprising:
 at least one first amylase, wherein the first amylase is an α-amylase from  Bacillus  sp. No. 707 or a functional fragment or a variant thereof; and   at least one second amylase, wherein the second amylase is an AA560 α-amylase from  Bacillus  sp. or a functional fragment or a variant thereof.   
     
     
         2 . The cleaning agent according to  claim 1 , wherein the first amylase has at least one amino acid substitution at one of positions 172, 202, 208, 255 and 261 in the count according to SEQ ID NO. 1. 
     
     
         3 . The cleaning agent according to  claim 1  wherein the second amylase has at least one amino acid substitution at one of positions 9, 26, 30, 33, 82, 37, 106, 118, 128, 133, 149, 150, 160, 178, 182, 186, 193, 195, 202, 203, 214, 231, 256, 257, 258, 269, 270, 272, 283, 295, 296, 298, 299, 303, 304, 305, 311, 314, 315, 318, 319, 320, 323, 339, 345, 361, 378, 383, 419, 421, 437, 441, 444, 445, 446, 447, 450, 458, 461, 471, 482 and 484 and/or one of the deletions D183* and G184* in the count according to SEQ ID NO. 2. 
     
     
         4 . The cleaning agent according to  claim 3 , wherein the second amylase, in the count according to SEQ ID NO. 2, includes amino acid substitutions at three or more of positions 9, 26, 149, 182, 186, 202, 257, 295, 299, 323, 339 and 345. 
     
     
         5 . The cleaning agent according to  claim 3 , wherein the second amylase includes the following amino acid substitutions and/or deletions in the count according to SEQ ID NO. 2:
 (i) M9L+M323T;   (ii) M9L+M202L/T/V/I+M323T;   (iii)M9L+N195F+M202L/T/V/I+M323T;   (iv)M9L+R118K+D183*+G184*+R320K+M323T+R458K;   (v) M9L+R118K+D183*+G184*+M202L/T/V/I+R320K+M323T+R458K;   (vi)M9L+G149A+G182T+G186A+M202L+T257I+Y295F+N299Y+M323T+A339S+E345R;   (vii) M9L+G149A+G182T+G186A+M202I+T257I+Y295F+N299Y+M323T+A339S+E345R;   (viii) M9L+R118K+G149A+G182T+D183*+G184*+G186A+M202L+T257I+Y295F+N299Y+R320K+M323T+A339S+E345R+R458K;   (ix)M9L+R118K+G149A+G182T+D183*+G184*+G186A+N195F+M202L+T257I+Y295F+N299Y+R320K+M323T+A339S+E345R+R458K;   (x) M9L+R118K+G149A+G182T+D183*+G184*+G186A+M202I+T257I+Y295F+N299Y+R320K+M323T+A339S+E345R+R458K;   (xi)M9L+R118K+D183*+D184*+N195F+M202L+R320K+M323T+R458K;   (xii) M9L+R118K+D183*+D184*+N195F+M202T+R320K+M323T+R458K;   (xiii) M9L+R118K+D183*+D184*+N195F+M202I+R320K+M323T+R458K;   (xiv) M9L+R118K+D183*+D184*+N195F+M202V+R320K+M323T+R458K;   (xv) M9L+R118K+N150H+D183*+D184*+N195F+M202L+V214T+R320K+M323T+R458K; or   (xvi) M9L+R118K+D183*+D184*+N195F+M202L+V214T+R320K+M323T+E345N+R458K.   
     
     
         6 . The cleaning agent according to  claim 1 , wherein the cleaning agent furthermore comprises at least one protease. 
     
     
         7 . The cleaning agent according to  claim 6 , wherein the at least one protease
 (a) comprises a subtilisin 309 from  Bacillus lentus  or a functional fragment or a variant thereof comprises an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 3 over the total length thereof and has at least one amino acid substitution at one of positions 9, 15, 66, 212 and 239 in the count according to SEQ ID NO. 3; and/or   (b) comprises a subtilisin 309 from  Bacillus lentus  or a functional fragment or a variant thereof, comprises an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 3 over the total length thereof and has an amino acid substitution at position 99 and an insertion of one amino acid between the amino acids at positions 99 and 100 in the count according to SEQ ID NO. 3; and/or   (c) comprises a protease from Bacillus alcalophilus PB92 or a functional fragment or a variant thereof, includes an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 4 over the total length thereof; and/or   (d) comprises an alkaline protease from  Bacillus lentus  DSM 5483 or a functional fragment or a variant thereof, includes an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 5 over the total length thereof; and/or   (e) comprises an amino acid sequence according to one of SEQ ID NOs: 6 to 10.   
     
     
         8 . The cleaning agent according to  claim 7 , wherein the at least one protease comprises two or more proteases according to  claim 7 . 
     
     
         9 . The cleaning agent according to  claim 1 , comprising a first amylase and a second amylase at a mass ratio of about 50:1 to about 1:50 (in each case based on the amount of active protein amylase 1 to amylase 2). 
     
     
         10 . The cleaning agent according to  claim 1 , wherein the weight fraction of each of the amylases based on the corresponding active protein, in the total weight of the agent is from about 1×10 −8  to about 5 wt. %. 
     
     
         11 . The cleaning agent according  claim 1 , comprising at least one further component selected from the group consisting of builders, surfactants, polymers, bleaching agents, bleach catalysts, bleach activators, non-protease and non-amylase enzymes, corrosion inhibitors, glass corrosion inhibitors, disintegrants, odorants and perfume carriers. 
     
     
         12 . The cleaning agent according to  claim 1 , wherein the cleaning agent is an automatic dishwashing agent, and the
 (1) automatic dishwashing agent is present in solid form; and/or   (2) the automatic dishwashing agent is present in preportioned form; and/or   (3) the automatic dishwashing agent comprises multiple compositions that are physically separated from each other.   
     
     
         13 . (canceled) 
     
     
         14 . Method for cleaning dishes in an automatic dishwasher, wherein a cleaning agent according to  claim 1  is dispensed into the interior of an automatic dishwasher while a dishwashing program is being executed, before the main washing cycle begins, or in the course of the main washing cycle. 
     
     
         15 . Use of a combination of a first amylase and a second amylase for improving the cleaning performance, of a cleaning agent, wherein:
 a) the first amylase is an α-amylase from  Bacillus  sp. No. 707 or a functional fragment or a variant thereof; and   b) the second amylase is an AA560 α-amylase from  Bacillus  sp. or a functional fragment or a variant thereof.   
     
     
         16 . The cleaning agent according to  claim 2 , wherein the first amylase has at least one amino acid substitution selected from the group consisting of M202L, M202V, M202S, M202T, M202I, M202Q, M202W, S255N and R172Q. 
     
     
         17 . The cleaning agent according to  claim 4 , wherein the second amylase, in the count according to SEQ ID NO. 2, includes amino acid substitutions and/or deletions at positions 118, 183, 184, 195, 320 and 458. 
     
     
         18 . The cleaning agent according to  claim 17 , wherein the second amylase, in the count according to SEQ ID NO. 2, has one or more of the following substitutions and/or deletions: R118K, D183*, G184*, N195F, R320K and/or R458K. 
     
     
         19 . The cleaning agent according to  claim 1 , wherein the first amylase includes an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 1 over the total length thereof. 
     
     
         20 . The cleaning agent according to  claim 1 , wherein the second amylase includes an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 2 over the entire length thereof. 
     
     
         21 . The cleaning agent according to  claim 6 , wherein the at least one protease comprises a protease from  Bacillus alcalophilus  PB92 or a functional fragment or a variant thereof, includes an amino acid sequence that is at least 80% identical to the amino acid sequence indicated in SEQ ID NO. 4 over the total length thereof, and that has at least one amino acid substitution at one of the following positions: 32, 33, 48 to 54, 58 to 62, 94 to 107, 116, 123 to 133, 150, 152 to 156, 158 to 161, 164, 169, 175 to 186, 197, 198, 203 to 216 in the count according to SEQ ID NO. 6,