Subtilase variants

Abstract

The present invention relates to novel subtilase variants exhibiting alterations 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 dishwash compositions, including automatic dishwash compositions.

1 - 17 . (canceled)  
     
     
         18 . A subtilase variant one or more of the following mutations in an amino acid sequence of a subtilase: 
 X11{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V}, deletion, insertion;    X30{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X31{R,N,D,Q,E,H,K,M,P,W,Y}, deletion, insertion;    X32{A,R,D,C,E,G,H,I,L,K,M,F,P,T,W,Y,v}, insertion;    X34{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V}, deletion;    X65{A,R,C,G,H,I,L,K,M,F,T,W,Y,V}, deletion, insertion;    X66{A,R,C,H,I,L,K,M,F,T,W,Y,V}, deletion, insertion;    X67{A,R,N,Q,G,H,I,K,M,F,P,S,T,W,Y,V};    X68{R,N,D,Q,E,G,H,I,L,K,F,P,S,T,W,Y,V}, deletion, insertion;    X69{A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X70G;    X71T;    X77N;    X83G, insertion;    X84V;    X85{A,R,N,D,Q,E,G,H,I,L,M,F,S,T,W,Y,V};    X90{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X95{R,K,F,W,Y,V}, deletion;    X107I    X110G, insertion;    X121V, insertion;    X122 insertion;    X123N;    X125S;    X150{A,R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X152{A,R,N,D,Q,E,H,K,F,P,W,Y,V};    X153{R,N,D,Q,E,G,H,I,L,K,M,F,T,W,Y,V};    X154{A,R,G,H,I,L,M,F,W,Y,V};    X164{A,R,H,T,W};    X165{R,L,F,W,Y};    X166{W,Y}, insertion;    X175{R,N,D,Q,E,G,H,K,M,F,P,T,W,Y,V};    X177{R,N,D,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X178{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X180{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,P,S,T,W,Y,V}, deletion, insertion;    X199{R,L,K,F,W,Y,V}, deletion, insertion;    X200{A,R,I,L,K,M,F,W,Y,V}, deletion, insertion;    X201{A,R,H,I,L,K,M,F,P,T,W,Y,V}, deletion, insertion;    X202{A,R,C,G,H,I,L,K,M,F,T,W,Y,V};    X207{A,R,N,C,Q,G,H,I,L,K,M,F,P,S,T,W,Y,V};    X220T, insertion;    X223A, insertion;    X225P, insertion;    X226H, insertion;    X227V, insertion;    X228A, insertion;    X229G, insertion;    X230 insertion;    X231A, insertion;    X253{A,R,N,D,Q,E,G,H,I,L,M,F,S,T,W,Y,V}, deletion, insertion;    X264{A,R,N,Q,G,H,I,L,M,F,S,T,W,Y,V}, deletion, insertion;    X266{A,R,N,D,C,Q,E,G,H,I,L,K,M,F,S,T,W,Y,V}, deletion, insertion;    V68A;    M175ML, M175MI    wherein the subtilase variant has protease activity and each position corresponds to a position of the amino acid sequence of subtilisin BPN′, shown in FIG. 1.    
     
     
         19 . The variant of  claim 18 , wherein the subtilase is a sub-group I-S1 subtilase.  
     
     
         20 . The variant of  claim 19 , wherein the subtilase is selected from the group consisting of subtilisin 1168, subtilisin BPN′, subtilisin DY, and subtilisin Carlsberg.  
     
     
         21 . The variant of  claim 18 , wherein the subtilase is a sub-group l-S2 subtilase.  
     
     
         22 . The variant of  claim 21 , wherein the parent subtilase is selected from the group consisting of subtilisin 147, subtilisin 309, subtilisin PB92, and subtilisin YaB.  
     
     
         23 . The variant of  claim 22 , wherein the variant is V11I; V11I+I44V; V11I+L96LA; S9R+V11I+A15T+T22A; V11I+L96LA+A108C+A138C; V11I+N76D+A194P+A230V; V11I+V841; V11I+V841+K251R; V11I+V301; V11I+V139L; V11I+V301; V301; V30I+V841; V30I+V139L; S9R+A15T+T22A+V301; V30I+V244R; V30I+K18594251R; V30I+V139L+Y167A+R170S+A194P+N218S; D32A; V68A; V68A+S106A; V68A,+V139I; V68A,+A158V; V68A+V203A; V68A+V139L; A48T+V68A+P131M; V51A+V68A+S106T+A168G; V51A+V68A+S106T+A168G; N76D+M175ML+A194P+A230V; N76D+M175MI+A194P+A230V.  
     
     
         24 . The variant of  claim 18 , which further comprises at least one further mutation at one or more of positions 27, 36, 56, 76, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 120, 123, 159, 167, 170, 206, 218, 222, 224, 232, 235, 236, 245, 248, 252 or 274.  
     
     
         25 . The variant of  claim 24 , wherein the variant further comprises S101 G+S103A+V104I+G159D+A232V+Q236H+Q245R+N248D+N252K.  
     
     
         26 . A cleaning or detergent composition, comprising a variant of  claim 18  and a surfactant.  
     
     
         27 . The composition of  claim 26 , which additionally comprises an amylase, cellulase, cutinase, lipase, oxidoreductase, another protease, or a mixture thereof.  
     
     
         28 . An isolated DNA sequence encoding a subtilase variant of  claim 18 .  
     
     
         29 . An expression vector comprising the isolated DNA sequence of  claim 28 .  
     
     
         30 . A microbial host cell transformed with the expression vector of  claim 29 .  
     
     
         31 . A microbial host cell of  claim 30 , which is a bacterium, preferably a Bacillus, especially a  B. lentus.    
     
     
         32 . A microbial host cell of  claim 30 , which is a fungus or yeast, preferably a filamentous fungus, especially an Aspergillus.  
     
     
         33 . A method for producing a subtilase variant, comprising 
 (a) culturing a host of  claim 30  under conditions conducive to the expression and secretion of the variant, and    (b) recovering the variant.