Protease Variants and Compositions

Abstract

The present invention relates to enzymes produced by mutating the genes for a number of subtilases and expressing the mutated genes in suitable hosts are presented. The enzymes exhibit improved wash performance in any detergent in comparison to their wild type parent enzymes.

1 . A subtilase enzyme variant having improved wash performance in detergents, comprising modification(s) in position(s) 252, 255 and/or 259 (in BASBPN numbering). 
     
     
         2 . A subtilase enzyme variant having improved wash performance in detergents, comprising at least one modification chosen from the group comprising (in BASBPN numbering):
 252 L+255I;   252V+255A;   252M+255C+259H;   252S+255E+259C;   252K+255S+259C; or   
       a variant comprising one or more conservative modification(s) in any of the above mentioned variants (e.g., a conservative modification of a 252L (a hydrophobic amino acid)+255I variant include variants such as 252I (a hydrophobic amino acid)+255I, and 252V (a hydrophobic amino acid)+255I. 
     
     
         3 . The subtilase enzyme variant according to  claim 2 , wherein the modification is chosen from the group comprising (in BASBPN numbering):
 N252L+T255I;   N252V+T255A;   N252M+T255C+S259H;   N252S+T255E+S259C;   N252K+T255S+S259C; or   
       a variant comprising one or more conservative modification(s) in any of the above mentioned variants (e.g., a conservative modification of a N252L (a hydrophobic amino acid)+T255I variant include variants such as N252I (a hydrophobic amino acid)+T255I, and N252V (a hydrophobic amino acid)+T255I. 
     
     
         4 . The variant of  claim 1 , wherein the parent subtilase is chosen from the sub-group I-S1. 
     
     
         5 . The variant of  claim 4 , wherein the parent subtilase is chosen from the group comprising ABSS168, BASBPN, BSSDY, and BLSCAR or functional variants thereof having retained the characteristic of sub-group I-S1. 
     
     
         6 . The variant of  claim 1 , wherein the parent subtilase is chosen from the sub-group I-S2. 
     
     
         7 . The variant of  claim 6 , wherein the parent subtilase is chosen from the group comprising BLS147, BLS309, BAPB92, TVTHER AND BYSYAB or functional variants thereof having retained the characteristic of sub-group I-S2. 
     
     
         8 . The variant of  claim 1 , wherein said modification(s) is/are combined with one or more modification(s) in any other position(s). 
     
     
         9 . The variant of  claim 8 , wherein said modification(s) is/are combined with modification(s) in one or more of the positions 27, 36, 57, 76, 97, 101, 104, 120, 123, 167, 170, 206, 218, 222, 224, 235 and 274. 
     
     
         10 . The variant of  claim 9 , wherein said subtilase belongs to the I-S2 sub-group and said further change is chosen from the group comprising K27R, *36D, S57P, N76D, G97N, S101G, V104A, V104N, V104Y, H120D, N123S, Y167A, Y167I, R170S, R170L, R170N, Q206E, N218S, M222S, M222A, T224S, K235L, and T274A. 
     
     
         11 . The variant of  claim 10 , comprising any of the variants V104N+S101G, K27R+V104Y+N123S+T274A, or N76D+V104A, or other combinations of these mutations (V104N, S101G, K27R, V104Y, N123S, T274A, N76D, V104A), in combination with any one or more of the substitutions, deletions and/or insertions mentioned in  claim 1 . 
     
     
         12 . The subtilase variant of  claim 1 , wherein said modification(s) is/are combined with modification(s) in one or more of the positions 129, 131, 133 and 194. 
     
     
         13 . The variant of  claim 12 , wherein said subtilase belongs to the I-S2 sub-group and said further change is chosen from the group comprising P129K, P131H, A133P, A133D and A194P. 
     
     
         14 . An isolated DNA sequence encoding a subtilase variant of any of  claims 1 . 
     
     
         15 . An expression vector comprising an isolated DNA sequence of  claim 14 . 
     
     
         16 . A microbial host cell transformed with an expression vector of  claim 15 . 
     
     
         17 . The microbial host of  claim 16 , which is a bacterium, preferably a  Bacillus , especially  B. lentus.    
     
     
         18 . The microbial host of  claim 16 , which is a fungus or yeast, preferably a filamentous fungus, especially an  Aspergillus.    
     
     
         19 . A method for producing a variant of  claim 1 , wherein a host of  claim 16  is cultured under conditions conducive to the expression and secretion of said variant, and the variant is recovered. 
     
     
         20 . A composition comprising a subtilase variant of  claim 1 . 
     
     
         21 . The composition according to  claim 20 , which additionally comprises a cellulase, lipase, cutinase, oxidoreductase, another protease, or an amylase. 
     
     
         22 . The composition of  claim 20 , wherein the composition is a detergent composition.