Methods of reducing odor

Abstract

The present invention relates to reduced odor. More specifically the invention relates to a method for hydrolyzing a lipase substrate comprising: adding to said substrate a lipase variant of SEQ ID NO: 3, which variant comprises a substitution at one or more positions corresponding to positions 4; 44; 45; 49; 58; 84; 88; 93; 95; 97; 104; 108; 131; 140; 147; 152; 158; 165; 176; 177; 187; 195; 199; 208; 211; 221; 222; 224; 232; 244; 247; 248; 249; 251; 255; and 259 of SEQ ID NO: 3, has lipase activity, and has at least 75% but less than 100% sequence identity to SEQ ID NO: 3 or a fragment thereof with lipase activity, in which method odor is reduced when compared to the method wherein SEQ ID NO: 3 is added to the lipase substrate. The present invention also relates lipase variants, polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of producing and using the variants.

1 . A method for hydrolyzing a lipase substrate comprising: adding to said substrate a lipase variant of SEQ ID NO: 3, which variant comprises one or more of the substitutions: D4H, D4K, D4R; V44Y; A45V, A451, A45L; Q49A, Q49G, Q49H, Q49K, Q49R, Q49S; I58K; I84P; I88A, I88P; F93L; P95V; N97D; A104A, A104G, A104S; K108A, K108G, K108S; R131A, R131G, R131S; T140A; A147G; S152A, S152G; H158Q; E165A, E165G, E165S; T176N; P177C, P177L, P1771, P177/V; K187N; H195D, H195S; I199G, I199V; F208G, F208K; L211C; K221A, K221G, K221P, K221S; D222P; S224G; I232H, I232K, I232N, I232R; F244T; V247A, V247G, V247S, V247D, V247L, V247P, V2471, V247Y; I248K, I248S; D2491, D249L, D249V; L251V, L2511, L251L; D255A, D255G, D255/S, G2591, G259L, and G259V of SEQ ID NO: 3, has lipase activity, and has at least 75% but less than 100% sequence identity to SEQ ID NO: 3 or a fragment thereof with lipase activity, in which method odor is reduced when compared to the method wherein SEQ ID NO: 3 is added to the lipase substrate. 
     
     
         2 . A method for lipid stain removal from a laundry, textile, fabric including house hold laundry washing, and Industrial laundry washing or hard surface cleaning including car wash, or industrial surface comprising: contacting said stain with a lipase variant of SEQ ID NO: 3, which variant comprises a substitution at one or more positions corresponding to positions: D4H, D4K, D4R; V44Y; A45V, A451, A45L; Q49A, Q49G, Q49H, Q49K, Q49R, Q49S; I58K; I84P; I88A, I88P; F93L; P95V; N97D; A104A, A104G, A104S; K108A, K108G, K108S; R131A, R131G, R131S; T140A; A147G; S152A, S152G; H158Q; E165A, E165G, E165S; T176N; P177C, P177L, P1771, P177/V; K187N; H195D, H195S; I199G, I199V; F208G, F208K; L211C; K221A, K221G, K221P, K221S; D222P; S224G; 232H, I232K, I232N, I232R; F244T; V247A, V247G, V247S, V247D, V247L, V247P, V2471, V247Y; I248K, I248S; D2491, D249L, D249V; L251V, L2511, L251L; D255A, D255G, D255/S, G2591, G259L, and G259V of SEQ ID NO: 3, has lipase activity, and has at least 75% but less than 100% sequence identity to SEQ ID NO: 3 or a fragment thereof with lipase activity, in which method odor is reduced when compared to the method wherein SEQ ID NO: 3 is added to the lipase substrate. 
     
     
         3 . (canceled) 
     
     
         4 . A lipase variant of SEQ ID NO: 3 comprising a substitution at one or more positions corresponding to positions: D4H, D4K, D4R; V44Y; A45V, A451, A45L; Q49A, Q49G, Q49H, Q49K, Q49R, Q49S; I58K; I84P; I88A, I88P; F93L; P95V; N97D; A104A, A104G, A104S; K108A, K108G, K108S; R131A, R131G, R131S; T140A; A147G; S152A, S152G; H158Q; E165A, E165G, E165S; T176N; P177C, P177L, P1771, P177/V; K187N; H195D, H195S; I199G, I199V; F208G, F208K; L211C; K221A, K221G, K221P, K221S; D222P; S224G; I232H, I232K, I232N, I232R; F244T; V247A, V247G, V247S, V247D, V247L, V247P, V2471, V247Y; I248K, I248S; D2491, D249L, D249V; L251V, L2511, L251L; D255A, D255G, D255/S, G2591, G259L, and G259V of SEQ ID NO: 3, wherein the variant has lipase activity and has at least 75% but less than 100% sequence identity to SEQ ID NO: 3 or a fragment thereof with lipase activity. 
     
     
         5 . The variant of  claim 4 , wherein the variant has at least sequence identity to SEQ ID NO: 3 or a fragment thereof with lipase activity. 
     
     
         6 . The variant of  claim 4 , comprising or constituting the substitutions selected from:
 a. V44Y;   b. A45V;   c. K221S;   d. D222P;   e. D255G;   f. H158Q+D255G;   g. S152A+D255G;   h. N97D+D255G;   i. Q49K+D255G;   j. R131A;   k. K108A;   l. A45V;   m. S152A+T176N+D255G;   n. Q49R;   o. Q49R+A104S+1232R;   p. Q49R+E165S+1232R;   q. Q49R+K187N+P189S+1232R;   r. Q49R+1232N;   s. Q49R+E165S+K187N+P189S+1232R;   t. Q49S+E165S+1232R;   u. Q49R+P177C+L211C+1232R; and   v. D4R+Q49R.   
     
     
         7 . The variant of  claim 4 , which further comprises one or more (e.g., several) substitutions at positions corresponding to positions 4, 27, 33, 38, 57, 58, 60, 83, 86, 91, 94, 97, 99, 111, 150, 163, 210, 216, 225, 227, 231, 233, 249, 254, 255, 256, 263, 264, 265, 266, 267, and 269 of SEQ ID NO: 3. 
     
     
         8 . The variant of  claim 4 , which variant has a reduced odor relative to SEQ ID NO: 3. 
     
     
         9 . A composition comprising the variant according to  claim 4 . 
     
     
         10 . A polynucleotide encoding the variant of  claim 4 . 
     
     
         11 . A nucleic acid construct comprising the polynucleotide of  claim 10 . 
     
     
         12 . An expression vector comprising the polynucleotide of  claim 10 . 
     
     
         13 . A host cell comprising the polynucleotide of  claim 10 . 
     
     
         14 . A method of producing a lipase variant of SEQ ID NO: 3, comprising: (a) cultivating the host cell of  claim 13  under conditions suitable for expression of the variant; and (b) recovering the variant. 
     
     
         15 . (canceled)