Lipase variants and polynucleotides encoding same
The present invention relates to a lipase variant of a parent lipase, which variant has lipase activity, at least 75% but less than 100% sequence identity to SEQ ID NO: 3 and comprises a substitution at one or more positions corresponding to positions 1; 2; 3; 4; 5; 6; 7; 9; 10; 11; 12; 16; 19; 30; 31; 34; 36; 37; 39; 40; 42; 44; 51; 52; 53; 54; 56; 58; 59; 70; 71; 72; 73; 83; 84; 86; 88; 90; 92; 93; 95; 96; 100; 101; 102; 104; 106; 109; 110; 112; 117; 119; 124; 125; 127; 128; 131; 132; 133; 134; 135; 137; 158; 159; 160; 161; 162; 163; 165; 166; 167; 168; 170; 181; 182; 183; 189; 190; 192; 194; 196; 202; 210; 211; 212; 220; 225; 227; 228; 229; 230; 231; 233; 237; 238; 239; 240; 242; 246; 247; 248; 252; 259; 262; 264; 269 of SEQ ID NO: 3. 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 which has lipase activity, at least 85% but less than 100% sequence identity to SEQ ID NO: 3, and comprises an amino acid substitution at one or more positions corresponding to positions 124; 182; 189; 190; 192; 230; and 96 (A, D,E,G,N,Q,R,S,W,Y) of SEQ ID NO: 3.
2 . The lipase variant of claim 1 , wherein the variant comprises an amino acid substitution at one or more positions corresponding to positions 96 (A, D,E,G,N,Q,R,S,W,Y); 124 (C,D,G,H,L,M,Q,R,S,V,W); 182 (A,C,F,G,I,L,R,S,T,V,W,Y); 189 (A,C,G,L,M,P,R,S); 190 (A,E,G,I,K,N,R,V,W); 192 (A,I,L,M,P,R,T,V); 230 (A,G,H,L,P,Q,R,S,T,V,W) of SEQ ID NO: 3.
3 . The variant of claim 1 selected from the group consisting of:
a. a polypeptide having at least 90% but less than 100% sequence identity to the polypeptide of SEQ ID NO: 3;
b. a polypeptide encoded by a polynucleotide having at least 90% but less than 100% sequence identity to the polypeptide coding sequence of SEQ ID NO: 3; and
c. a fragment of the polypeptide of SEQ ID NO: 3, wherein the fragment has lipase activity.
4 . The lipase variant of claim 1 , wherein the variant has one or more improved properties relative to the polypeptide of SEQ ID NO: 3 measured as an Improvement Factor (IF) that is greater than 1.0, wherein the improved properties are selected from the group consisting of: Thermostability; Thermostability in Detergent; Hydrolytic Activity in Detergent; and Stability in Detergent.
5 . The lipase variant of claim 1 , wherein the Improvement Factor (IF) is greater than 1.0 in at least one of the assays selected from the group consisting of: A Thermostability Assay; B Thermostability in Detergent Assay; C Performance Screening Assay; and D Stability in Detergent Assay.
6 . The variant of claim 5 , wherein the Improvement Factor (IF) is greater than 1.0 in at least two assays.
7 . The variant of claim 5 , wherein the Improvement Factor (IF) is at least 1.1.
8 . The variant of claim 1 , wherein the number of substitutions are 1-40.
9 . The lipase variant of claim 1 , which further comprises an amino acid substitution at one or more positions corresponding to positions: A8T; Y28LC; 141T; C43S; D48E; K50E; Y60AC; D61NKRAVLSTC; R80C; S84C; I86C; N87C; A90C; D91NKRAVLST; F94LTK; S98P; S103R; D113NKRAVLST; S114C; G116V; N120KD; K131F; G156D; N186Y; E201QKRAVLST; I204VATSG; V205IL; L208VATSG; F213VATSG; H217N; D226NKRAVLST; T236A; T241S; D243NKRAVLSTH; V254IATSG; L255VATSG; D256NKRAVLST; L258VATSG; Y260WCGV; and L267VATSG of SEQ ID NO: 3.
10 . A method of hydrolyzing a lipase substrate, comprising contacting said substrate with the lipase variant of claim 1 .
11 . A composition comprising the lipase variant of claim 1 .
12 . A method of cleaning, comprising contacting a surface or an item with the lipase variant of claim 1 .
13 . A polynucleotide encoding the lipase variant of claim 1 .
14 . A nucleic acid construct comprising the polynucleotide of claim 13 .
15 . An expression vector comprising the polynucleotide of claim 13 .
16 . A host cell comprising the polynucleotide of claim 13 .
17 . A method of producing a lipase variant, comprising:
a. cultivating the host cell of claim 16 under conditions suitable for expression of the lipase variant; and
b. recovering the lipase variant.
18 . The variant of claim 1 , wherein the variant has at least 92% but less than 100% sequence identity to SEQ ID NO: 3.
19 . The variant of claim 1 , wherein the variant has at least 95% but less than 100% sequence identity to SEQ ID NO: 3.