Lipase variants and polynucleotides encoding same

Abstract

The present invention relates to lipase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

1 . A lipase variant, comprising a substitution at positions corresponding to position 210 which is not a negatively charged amino acid, and position 255 which is not I, and wherein position 256 is not K, of SEQ ID NO: 2, and wherein the variant has lipase activity and has at least 60% but less than 100% sequence identity to SEQ ID NO: 2. 
     
     
         2 . The lipase variant of  claim 1 , wherein the variant further comprises a substitution at positions corresponding to position 33, 91, 231 and/or 233. 
     
     
         3 . The lipase variant of  claim 1 , wherein the substitution at 33 is K or Q; 91 is Q, 210 is A, C, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y; 231 is H, K, or R; 233 is H, K, or R; 255 is A, C, D, E, F, G, H, K, L, M, N, P, Q, R, S, T, V, W, or Y; and 256 is A, C, D, E, F, G, H, I, L, M, N, P, Q, R, S, T, V, W, or Y. 
     
     
         4 . The lipase variant of  claim 1  selected from the group consisting of:
 a. a polypeptide having at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 2; 
 b. a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, or (ii) the full-length complement of (i); 
 c. a polypeptide encoded by a polynucleotide having at least 80% identity to the mature polypeptide coding sequence of SEQ ID NO: 1; and 
 d. a fragment of the mature polypeptide of SEQ ID NO: 2, which has lipase activity. 
 
     
     
         5 . The lipase variant of  claim 1 , comprising or consisting of substitutions at two positions corresponding to positions 210+255, three positions corresponding to positions 210+231+255; 210+233+255; 210+255+256; or 91+210+255; four positions corresponding to positions 33+91+210+255; 210+231+233+255, 210+231+255+256; or 210+233+255+256; or five positions corresponding to positions 210+231+233+255+256 of SEQ ID NO: 2. 
     
     
         6 . The lipase variant of  claim 1 , comprising or consisting of the set of substitutions selected from:
 a. N33Q+G91Q+E210Q+I255A;   b. D27R+N33K+G38A+F51V+D96E+K98E+D111A+G163K+E210K+D254S+I255G+P256T;   c. D27R+G38A+F51V+D96E+K98E+D111A+G163K+E210K+D254S+I255G+P256T;   d. D27R+N33K+G38A+F51V+S54T+E56K+L69R+D96E+K98E+D111A+G163K+E210K+Y220F+D254S+I255G+P256T:   e. D27R+G38A+F51V+L69R+D96E+K98E+D111A+G163K+E210K+D254S+I255G+P256T; and   f. Q4V+S58N+V60S+E210K+I255F   
     
     
         7 . The lipase variant of  claim 1 , further comprising one or more (e.g., several) of the substitutions N33Q; G91Q; T143A; and A150G. 
     
     
         8 . The lipase variant of  claim 1 , which has a reduced level or low level of odor as compared with SEQ ID NO: 2. 
     
     
         9 . A composition comprising the lipase variant of  claim 1 . 
     
     
         10 . A method of cleaning a surface, comprising contacting the surface with the composition of  claim 9 . 
     
     
         11 . A polynucleotide encoding the lipase variant of  claim 1 . 
     
     
         12 . A nucleic acid construct comprising the polynucleotide of  claim 11 . 
     
     
         13 . An expression vector comprising the polynucleotide of  claim 11 . 
     
     
         14 . A host cell comprising the polynucleotide of  claim 11 . 
     
     
         15 . A method of producing a lipase variant, comprising:
 a. cultivating the host cell of  claim 14  under conditions suitable for expression of the lipase variant; and   b. recovering the lipase variant.