Subtilase variants

Abstract

The present invention relates to novel subtilase variants exhibiting improvements relative to the parent subtilase in one or more properties including: wash performance, thermal stability, storage stability or catalytic activity. The variants of the invention are suitable for use in e.g., cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.

1 . A subtilase variant comprising one or more sets of modifications selected from the group consisting of 
 T143K, Y167A, R170S, A194P;    Y167A, R170S, A194P, K251 R;    Y167A, R170S, A194P, S265K;    Y167A, R170S, A194P, V244R;    S141E, Y167A, R170S, A194P;    Y167A, R170S, M1751;    Y167A, R170S, A172T;    Y167A, R170S, A174V, M175F;    Y167A, R170S, A172V, A174V;    Y167A, R170S, A172E;    Y167A, R170S, M175L;    Y167A, R170S, A174T;    Y167A, R170S, A174T, M175L;    G53C, G61E;    A98S, S99D, G100S;    S9R, T22A, V68A, S99A, *99aD;    S9R, P14H, R19L, N62D;    G61P, *99aS;    N43S, N62D;    *96aG, P131S, V203A, A228T;    N62D, A232C, Q236L, Q245N;    *96aA, A98T, R247K;    S99D, S101R, S103A, V104I, G160S, A194P, L217D;    *61aD;    N62D, S106A;    V68A, S106M, N184D;    S9R, A15T, *97aV, H120N;    A15M, A16P, *99aD;    *99aE, G160S, S163T, G195S, G211S, K237R, G258A, T260L;    G23S, *99aD, A194P, S242T, Q245R;    G100S, N173D;    Y167A, R170S, A172E;    A98T, Q137L, Y167A, R170S, M175L;    *98aA, S99D;    S99A, *99aD, V203A;    N62D, K237R;    V11M, N76D, L126F, K251R;    S9F, A15L, A16P, T22I, *98aA, S99D, R170H;    *96aA, *130aG, P131H;    E54D, N62D;    *98aA, *98bS, S99G, S101T;    S9R, A15T, V68A, I79T, G102S, P131H, Q137H;    *100aA, *100bG, *100cS, *100dG;    V68A, L111I;    *98aA, R170H, Q245R;    I35V, N62D, N183D, T224S;    *97aG, P131S, V203A, A228T;    S9R, R10K, P14Q, T22A, Y167A, R170S;    S9R, *22aL, S57A, G61E, *98aA, V139L, N173S;    P14T, N18K, Y167A, R170S;    S9R, Q12E, P14Q, K27R, Y167A, R170S;    N62D, R170L;    N62D, R170S, Q245R;    Y167A, R170S, A194P, K251R, S265K;    P14T, N18K, Y167A, R170S, A194P;    N62D, A151G, K237R;    N62D, A151G, Q245R;    N62D, A151G, K237R, Q245R;    S103A, V104I, G159D, A232V, Q236H, Q245R;    S9R, A15T, T22A, V139L;    S9R, A15T, G61E, A85T, E89Q, P239L, Q245C;    S9R, A15T, V68A, H120N, Q245R;    N248R;    S9R, A15T, *22aL, V139L, N204D, Q245L;    N218S;    S9R, A15T, V68A, Q245R, N252K;    S9R, A15T, V68A, Q245R, H120N;    V68A, S106A, H120N;    V68A, S106A, N252K;    A15T, V68A, S99G, Q245R, N261D;    S9R, V68A, S99G, Q245R, N261D;    V68A, S99G, Q245R, N261D;    S9R, A15T, V68A, S99G, N261D;    S9R, A15T, V68A, Q245R, N261D;    S9R, A15T, *22aL, V139L, S163G, N204D, Q245L;    Q245R, N252H;    S9R, *22aL, G61 E, *97aA, M119I, Q137H, N173S;    V68A, S106A, T213A;    S9R, A15T, V68A, H 120 N, P131S, Q137H, Q245M;    S9R, A15T, V68A, 172F, S99G, Q245R, N261D;    S9R, A15T, V68A, S99D, Q245R, N261D;    S9R, A15T, V68A, S99G, A194P, Q245R, N261D;    S9R, A15T, V68A, N76I, S99G, Q245R, N261D and    S9R, A15T, V68A, S99G, A228V, Q245R, N261D.    
     
     
         2 . A subtilase variant according to  claim 1 , wherein said variant further comprises one or more of the modifications K27R, *36D, S56P, N62D, V68A, N76D, S87N, G97N, S99SE, S101G, S101R, S103A, V104A, V104I, V104N, V104Y, S106A, H120D, H120N, N123S, G159D, Y167A, R170S, R170L, A194P, N204D, V2051, Q206E, L217D, N218S, N218D, M222S, M222A, T224S, A232V, K235L, Q236H, Q245R, N248D, N252K, T274A, S101G+V104N, S87N+S100G+V104N, K27R+V104Y+N123S+T274A, N76D+S103A+V104I, S99D+S101R+S103A+V104I+G160S, S3T+V4I+S99D+S101R+S103A+V104I+G160S+V199M+V205I+L217D, S3T+V4I+S99D+S101R+S103A+V104I+G160S+A194P+V199M+V205I+L217D, S3T+V4I+S99D+S101R+S103A+V104I+G160S+V205I and N76D+V104A.  
     
     
         3 . A subtilase variant according to  claim 1  comprising the following substitutions: S101 G+S103A+V104I+G159D+A232V+Q236H+Q245R+N248D+N252K.  
     
     
         4 . The variant according to  claim 1 , wherein the parent subtilase belongs to the sub-group I-S1.  
     
     
         5 . The variant according to  claim 1 , wherein the parent subtilase belongs to the sub-group I-S2, and wherein the parent preferably is BLSAVI.  
     
     
         6 . A cleaning or detergent composition, comprising a variant of  claim 1  and a surfactant.  
     
     
         7 . A composition of  claim 6 , which additionally comprises a cellulase, a lipase, an amylase, a cutinase, a protease, a hemicellulase, an esterase, a lactase, a glycoamylase, a polygalacturonase, a beta-galactosidase, a ligninase, or a mixture thereof.  
     
     
         8 . An isolated DNA sequence encoding a subtilase variant of  claim 1 .  
     
     
         9 . An expression vector comprising the isolated DNA sequence of  claim 8 .  
     
     
         10 . A microbial host cell transformed with the expression vector of  claim 9 .  
     
     
         11 . A microbial host cell of  claim 10 , which is a bacterium.  
     
     
         12 . A microbial host cell of  claim 10 , which is a fungus or yeast.  
     
     
         13 . A method for producing a subtilase variant, comprising 
 (a) culturing a host of  claim 1  under conditions conducive to the expression and secretion of the variant, and    (b) recovering the variant.