Lipase variants and polynucleotides encoding same

Abstract

The present invention relates to a variant of a parent lipase, which parent lipase has the amino acid sequence of SEQ ID NO: 2, wherein the variant has lipase activity, comprises a substitution at a position corresponding to position 51 and comprises a substitution or remains unaltered at a position corresponding to position 96 of SEQ ID NO: 2. 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 variant of a parent lipase, wherein the variant has lipase activity, comprises a substitution at a position corresponding to position 51 of SEQ ID NO: 2, and comprises a substitution or remains unaltered at a position corresponding to position 96 of SEQ ID NO: 2. 
     
     
         2 . The variant of  claim 1 , wherein the substitution at a position corresponding to position 51 of SEQ ID NO: 2 is I, L, M, V or Y. 
     
     
         3 . The variant of  claim 1  selected from the group consisting of:
 a. a polypeptide having at least 60%, but less than 100% sequence identity to the parent lipase. 
 b. a fragment of the polypeptide of the parent lipase, wherein said fragment has lipase activity. 
 
     
     
         4 . The variant of  claim 1 , wherein the parent lipase has the amino acid sequence of SEQ ID NO: 2; SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         5 . The variant of  claim 1  which comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen or seventeen substitutions selected from the group consisting of: E56H/K/R; N33H/K/R/P/Q; E210H/K/R; S54T; T64S; K98E/D/I/R/T/V; E87E/F/H/I/K/M/Q/R; V2F/G/H/K/R/Y; L269I/V; D57G/N; L69H/K/R; V176L; W260F/Y; K163G/P; A243P/S/T; Y220F; and N101D/E. 
     
     
         6 . The variant of  claim 1  comprising a set of substitutions selected from the following group consisting of:
 a. N33K+F51V+E56K+L69R+K98E+E210K; 
 b. N33K+F51V+E56K+K98E+K163P+E210K; 
 c. N33K+F51V+E56K+L69R+K98E+V176L+E210K+Y220F; 
 d. N33K+F51V+E56K+L69R+K98E; 
 e. F51V+E56K+E87K+K98E; 
 f. N33K+F51V+E56K+K98E+V176L+E210K+Y220F; 
 g. N33K+F51V+K98E; 
 h. N33K+F51V+E56K+L69R+K98E+V176L+E210K+W260Y; 
 i. F51V+L69R+K98E+E210K; 
 j. N33K+F51V+S54T+E56K+L69R+K98E+W260Y; 
 k. F51V+E87K+K98E; 
 l. N33P+F51V+K98E; 
 m. N33K+F51V+S54T+E56K+K98E+E210K+W260Y; 
 n. F51V+D57G+N101D+E210K; 
 o. N33K+F51V+K98E+E210R+W260Y; 
 p. N33K+F51V+S54T+L69R+E87K+K98E+E210K+Y220F; 
 q. F51V+K98E+N101D; 
 r. N33K+F51V; 
 s. F51V; 
 t. V2K+N33K+F51V+S54T+L69R+K98E+V176L+E210K+W260Y; 
 u. N33K+F51V+S54T+K98E+W260Y; 
 v. N33K+S54T; 
 w. N33K+L69R+K98E+K163P; 
 x. N33K+K98E; 
 y. F51L; 
 z. E87K+A243S; 
 aa. N33K; 
 bb. E210R; 
 cc. S54T; 
 dd. E56K; 
 ee. T64S; 
 ff. E87K; 
 gg. V2K; 
 hh. N33P; 
 ii. L269V; 
 jj. D57G; 
 kk. E210K; 
 ll. V2Y; 
 mm. L69R; 
 nn. K98T; 
 oo. V176L; 
 pp. W260Y; 
 qq. K98I; 
 rr. K163P; 
 ss. E210K+A243S; 
 tt. K98E; 
 uu. V2Y+N33K+F51V; 
 vv. V2K+F51V; 
 ww. V2K+N33K+F51V; 
 xx. N33K+F51V+E56K+L69R+E96D+K98E; 
 yy. N33K+L69R+E96D+K98I; 
 zz. N33K+F51V+S54T+E56K+K98I; 
 aaa. S54T+E96D+K98T+K163P+V176L+W260Y+L269V; 
 bbb. F51I; 
 ccc. N33P F51V; 
 ddd. F51V L69R E87K K98E; 
 eee. N33P F51V D57G E210K; 
 fff. N33K F51V D57G K98I; 
 ggg. V2K+N33K+F51V+V187P; 
 hhh. V2K+N33K+F51V+S224F; 
 iii. V2K+N33K+F51V+L227D; 
 jjj. V2K+N33K+F51V+V228P; 
 kkk. V2K+N33K+F51V+L69R+K98E+V176L+E210K+L227G; 
 lll. V2K+N33K+F51V+L69R+K98E+V176L+E210K+S224F+L227G; 
 mmm. V2K+N33K+F51V+L69R+K98E+V176L+E210K+L227G+V228P; 
 nnn. V2K+N33K+F51V+L69R+K98E+V176L+V187P+E210K+L227G; 
 ooo. N33K+F51V+S54T+E56K+K98I+T143A; 
 ppp. V2Y+N33K+F51V+E56K+L69R+E96D+K98E; 
 qqq. N33K+F51V+E56K+L69R+E96D+K98E+V187D; 
 rrr. N33K+F51V+E56K+L69R+E96D+K98E+G225P; 
 sss. N33K+F51V+K98I+H198S+Y220F 
 nnn. N33K F51V L93I N94R F95V 
 uuu. F51V I86P E87A G91A N92D F95Y D96T L97P K98Q T2521 T2551 
 vvv. A243S; 
 www. F51V+V176L; 
 xxx. F51V+K98E+E210K; 
 yyy. L69R+K98E+E210K; 
 zzz. N33K+E56K+L69R+K98E; 
 aaaa. N33K+F51L; 
 bbbb. N33K+F51V+E56K+L69R+K98E+T256P; 
 cccc. N33K+F51V+S54T+K98I; 
 dddd. V2K+N33K; 
 yyy. V2K+N33K+L69R+K98E+V176L+E210K+L227G; 
 zzz. N33K+F51V+K98I+Y220F; 
 æææ. N33K+F51V+L69R+K98E; 
 ØØØ. N33K+F51V+K98I; 
 ååå. V2Y+N33K+F51V+E56K+L69R+K98E; 
 aaaa. F51V+E56K+L69R+K98E; 
 bbbb. V2K+N33K+F51V+L69R+K98E+E210K+L227G; 
 cccc. N33K+F51V+E56K+L69R+K98E+G225P; 
 mmmm. N33K+F51V+E56K+L69R+K98E+V187D; and 
 eeee. N33K+F51V+L69R+K98E+V176L+E210K+L227G; 
 
     
     
         7 . The variant of  claim 1 , which further comprises an alteration at positions corresponding to any of positions: 4, 27, 33, 38, 57, 58, 60, 83, 86, 91, 94, 97, 99, 111, 150, 163, 210, 216, 225, 227, 231, 249, 254, 255, 256, 263, 264, 265, 266, 267, and 269 of SEQ ID NO: 2. 
     
     
         8 . The variant of  claim 1 , which has improved stability as compared to the parent lipase. 
     
     
         9 . The variant of  claim 8 , wherein the stability is stability in detergent compositions, stability in detergent compositions comprising protease, stability in the presence of protease, stability under storage conditions, stability under storage conditions in the presence of protease, and/or thermostability. 
     
     
         10 . A detergent composition comprising the variant of  claim 1 . 
     
     
         11 . (canceled) 
     
     
         12 . A polynucleotide encoding the variant of  claim 1 . 
     
     
         13 . A nucleic acid construct comprising the polynucleotide of  claim 12 . 
     
     
         14 . An expression vector comprising the polynucleotide of  claim 12 . 
     
     
         15 . A host cell comprising the polynucleotide of  claim 12 . 
     
     
         16 . A method of producing a lipase variant, comprising: (a) cultivating the host cell of  claim 15  under conditions suitable for expression of the variant; and (b) recovering the variant. 
     
     
         17 . A method for obtaining the variant of  claim 1 , comprising introducing into a parent lipase a substitution at a position corresponding to position 51 and optionally introducing a substitution at a position corresponding to position 96 of SEQ ID NO: 2.