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(51) International Patent Classification: (21) International Application Number: PCT/EP2020/068459 (22) International Filing

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(51) International Patent Classification: (21) International Application Number: PCT/EP2020/068459 (22) International Filing ( (51) International Patent Classification: C12N 9/20 (2006.01) C12N 15/52 (2006.01) CUD 3/386 (2006.01) (21) International Application Number: PCT/EP2020/068459 (22) International Filing Date: 0 1 July 2020 (01.07.2020) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 19183918.2 02 July 2019 (02.07.2019) EP (71) Applicant: NOVOZYMES A/S [DK/DK]; Krogshoejvej 36, 2880 Bagsvaerd (DK). (72) Inventors: CHRISTENSEN, Sune, Fang; Krogshoe¬ jvej 36, 2880 Bagsvaerd (DK). BAUNSGAARD, Lone; Krogshoejvej 36, 2880 Bagsvaerd (DK). FULTON, Alexander, David,; Krogshoejvej 36, 2880 Bagsvaerd (DK). SANDSTROM, Anders, Gunnar,; Krogshoejvej 36, 2880 Bagsvaerd (DK). (81) Designated States (unless otherwise indicated, for every kind of national protection available) : AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, IT, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, WS, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available) : ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, Cl, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Published: — with international search report (Art. 21(3)) — before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) — with sequence listing part of description (Rule 5.2(a)) (54) Title: LIPASE VARIANTS AND COMPOSITIONS THEREOF (57) Abstract: The present invention relates to lipase variants of the parent lipase shown as SEQ ID NO: 2 with improved wash performance. The invention also relates to polynucleotides encoding the lipase variants of the invention, compositions comprising a lipase variant of the invention as well as methods of producing such lipase variants of the invention and the use of the lipase variants or compositions thereof. LIPASE VARIANTS AND COMPOSITIONS THEREOF REFERENCE TO A SEQUENCE LISTING This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference. BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to lipase variants, polynucleotides encoding the lipase variants, compositions comprising lipase variants as well as methods of producing said lipase variants and the use of lipase variants or compositions thereof. Description of the Related Art Lipases are important biocatalysts which have shown to be useful for various applications. Lipases have been commercialized as active ingredient in detergent compositions for the removal of lipid stains by hydrolyzing triglycerides to generate fatty acids. Current cleaning and/or fabric care compositions comprise many ingredients which are interfering with the performance and thus the ability of lipases to remove lipid stains. WO 1996/013578 relates to detergent composition comprising lipolytic enzymes derived from strains of the genus Absidia. WO 1997/027276 relates to nucleic acid sequences encoding polypeptides having Absidia lipase activity. WO 2017/005816 concerns Absidia lipase variants with improved stability. WO 2017/005798 concerns Absidia lipase variants with reduced odor generation during lipid stain removal. Accordingly, the need exists for Absidia lipases that can function in the harsh environment of compositions used for cleaning. There is a need and desire for lipases and lipase variants with improved wash performance. SUMMARY OF THE INVENTION The present invention provides lipase variants with improved properties compared to its parent, preferably the lipase shown as SEQ ID NO: 2 . The improved properties may be one or more selected from: increase in thermostability; increase in thermostability in detergent; increased in hydrolytic activity in detergent; and increase in stability in detergent. The present invention provides lipase variants including but not limited to variants that are particularly useful in various cleaning applications. The invention also provides methods of cleaning using the lipase variants of the present invention. A lipase variant of the invention is useful for cleaning. The variants are achieved by introducing modifications, such as substitutions into a parent lipase, in particular the parent lipase shown as SEQ ID NO: 2 . The present invention relates to improved lipase variants and composition comprising a lipase variant of the invention. In the first aspect, the invention relates to variants of a parent lipase, wherein: i) the variant is a polypeptide having lipase activity; ii) the variant is a polypeptide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity to the polypeptide shown as SEQ ID NO: 2 ; iii) the variant is a polypeptide encoded by a polynucleotide having at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, but less than 100% sequence identity to the mature polypeptide coding sequence shown as SEQ ID NO: 1 ; iv) the variant is a fragment of the polypeptide of ii) or iii) that has lipase activity; wherein the variant comprises: (a) a substitution corresponding to D4A; I7D; K14E; T17S; G34D; R36E; R36M; R36V; R36Q; S38D; S38E; G43D; G43H; N47D; N47E; N47G; N47H; Q49T; Q49V; L57S; D60G; D60V; G68I; E69M; E69T; K70D; K70H; K70Q; A89Y; V94D; V94L; N102M; G109R; Q 118S; K120A; K120D; K120E; K120N; K120M; K120G; K120P; K120Q; K120S; K120V; A123E; K136G; H158E; H158Q; H161D; H161 E ; A162S; N163E; N163H; I174A; P177G; P177I; P177K; P177L; A178I; A178L; A178T; A178V; N181 K ; N181 L ; N181T; N181W; Y182W; V183M; 1184V; G185A; G185I; G185K; G185R; G185T; G185W; G185Y; K187G; K187L; P189A; P189L; P189M; P189N; P189Q; P189R; P189T; P189V; P189W; I199T; 1199V; E196M; S206G; S206V; A207N; L21 1C; R223A; R223E; R223G; R223H; R223K; R223N; R223Q; S224T; I232P; 232Q; T234S; D235A; I248L; S252N; L260F; and/or (b) substitutions corresponding to: N163E+R223N; K70E+S206G; R101V+S206V; A15E+K120Q; D130R+S206G; D130R+S206V; T2S+A15E; and/or (c) substitutions corresponding to R36V+V 183M+R223N ; N47H+K70E+S206G; N163E+R223N+S252N; N47D+K120G+R223N; G43D+N47D+K120G; K70E+H195S+S206G; K70E+L156F+S206G; A178Y+P189V+R223N; R36E+G43D+N47D; K70D+N163E+S252N; N47D+K120G+N163E; K70E+P189Q+S206G; R36T+K70E+S206G; R36I+K70E+S206G; Q49T+K70E+S206G; G43D+K70E+S206G; R36K+K70E+S206G; K70E+P189M+S206G; K52A+K70E+S206G; R36M+K70E+S206G; R36E+K70E+S206G; R101V+S206V+R223S; R36V+N47D+K120G; K14A+K70E+S206G; D4C+S206G+N236C; K70E+G185L+S206G; R36Y+K70E+S206G; R36L+K70E+S206G; K70E+P189L+S206G; R36A+K70E+S206G; R101 V+D130R+S206G; D4C+S206G+N236C; D4C+S206V+N236C; R36F+K70E+S206G; K70E+K120P+S206G; and/or (d) substitutions corresponding to N163E+G 185H+R223N+S252N; N163E+N1 8 1W+R223N+S252N ; K70D+N163E+R223N+S252N; D4C+S206G+N236C+S252N; D4C+S206G+R223N+N236C; A178Y+V183M+P189V+R223N; and/or (e) substitutions corresponding to D4C+P1 89V+S206G+R223N+N236C; D4C+N163E+S206G+N236C+S252N; D4C+N163E+S206G+R223N+N236C; D4C+A178Y+S206G+R223N+N236C; D4C+K1 08A+D1 30R+S206V+N236C; and/or (f) substitutions corresponding to S1C+A8C+T17C+L225R+N230C+L254C; S1C+A8C+T17C+L225R+N230C+L254C; D4C+N97D+R101V+D130R+S206V+N236C; and/or (g) substitutions corresponding to S1C+A8C+T17C+I184N+L225C+N230C+L254C; S1C+A8C+T17C+Y75F+L225C+N230C+L254C; S1C+A8C+T17C+Q49L+L225C+N230C+L254C; S 1C+A8C+T1 7C+Y1 67F+L225C+N230C+L254C; S1C+A8C+T17C+Y190F+L225C+N230C+L254C; S1C+A8C+T17C+Q49I+L225C+N230C+L254C; S1C+A8C+T17C+L225C+N230C+F244N+L254C; S1C+A8C+A15E+T17C+L225C+N230C+L254C; S1C+A8C+T17C+A162S+L225C+N230C+L254C; S1C+A8C+T17C+V183S+L225C+N230C+L254C; S 1C+A8C+T1 7C+1 164L+L225C+N230C+L254C; S1C+A8C+T17C+Y19F+L225C+N230C+L254C; S1C+A8C+T17C+E165S+L225C+N230C+L254C; S1C+A8C+T17C+Q49S+L225C+N230C+L254C; S1C+A8C+T17C+K187N+L225C+N230C+L254C; S1C+A8C+A15R+T17C+L225C+N230C+L254C; and/or (h) substitutions corresponding to R101 V+A131 R+A152S+N176T+S206G+R223S+F247V+G255D; S1C+D4N+R6S+A8C+T17C+L225C+N230C+L254C; R101 V+A131 R+A152S+N176T+S206G+R223S+F247V+G255D; S1C+A8C+T17C+K136N+V138S+L225C+N230C+L254C; A26C+I32C+A45V+Q49K+N97D+D130R+H159K+S224R; S1C+A8C+T17C+K70E+I184N+L225C+N230C+L254C S1C+A8C+T17C+V76I+L225C+N230C+L254C; and/or (i) substitutions corresponding to S1C+A8C+T17C+H159K+H161 N+N163S+L225C+N230C+L254C; S1C+A8C+T17C+V76I+V138L+I164L+L225C+N230C+L254C; and/or (j) substitutions corresponding to S 1C+A8C+T1 7C+S38T+V76I+V1 38L+1 164L+L225C+N230C+L254C; and/or (k) substitutions corresponding to A8C+T17C+R36N+G43D+Q49T+K70N+K72S+K120G+S206V+R223N+L225C+L254C; and/or (L) substitutions corresponding to A8C+T17C+R36V+S38D+G43D+N47D+Q49N+K70N+K72S+K120G+T148V+E165S+N181W+ S206V+R223H+L225C+L254C; of the polypeptide shown as SEQ ID NO: 2 .
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