CN101960008A - 具有脂肪酶活性的多肽和编码该多肽的多核苷酸 - Google Patents
具有脂肪酶活性的多肽和编码该多肽的多核苷酸 Download PDFInfo
- Publication number
- CN101960008A CN101960008A CN2009801069853A CN200980106985A CN101960008A CN 101960008 A CN101960008 A CN 101960008A CN 2009801069853 A CN2009801069853 A CN 2009801069853A CN 200980106985 A CN200980106985 A CN 200980106985A CN 101960008 A CN101960008 A CN 101960008A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- seq
- change
- lipase
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 143
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 140
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 140
- 235000019626 lipase activity Nutrition 0.000 title claims description 23
- 102000040430 polynucleotide Human genes 0.000 title claims description 15
- 108091033319 polynucleotide Proteins 0.000 title claims description 15
- 239000002157 polynucleotide Substances 0.000 title claims description 15
- 150000004666 short chain fatty acids Chemical class 0.000 claims abstract description 5
- 102200142011 rs121909050 Human genes 0.000 claims description 55
- 102220238245 rs1555952639 Human genes 0.000 claims description 53
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims description 38
- 238000005406 washing Methods 0.000 claims description 26
- 102220126864 rs147455726 Human genes 0.000 claims description 25
- 150000001413 amino acids Chemical class 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 20
- 102000039446 nucleic acids Human genes 0.000 claims description 17
- 108020004707 nucleic acids Proteins 0.000 claims description 17
- 150000007523 nucleic acids Chemical class 0.000 claims description 17
- 239000013604 expression vector Substances 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 13
- 238000003259 recombinant expression Methods 0.000 claims description 12
- 102220079878 rs78870822 Human genes 0.000 claims description 12
- 102220539256 Programmed cell death 1 ligand 2_S58T_mutation Human genes 0.000 claims description 9
- 102220031962 rs431825177 Human genes 0.000 claims description 9
- 102220479102 CD59 glycoprotein_N33Q_mutation Human genes 0.000 claims description 7
- 102220466467 Testis-specific H1 histone_D27N_mutation Human genes 0.000 claims description 7
- 238000009472 formulation Methods 0.000 claims description 7
- 230000007062 hydrolysis Effects 0.000 claims description 7
- 238000006460 hydrolysis reaction Methods 0.000 claims description 7
- 150000002632 lipids Chemical class 0.000 claims description 6
- 239000005711 Benzoic acid Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 102220642347 PTB domain-containing engulfment adapter protein 1_I86S_mutation Human genes 0.000 claims description 3
- 102220582968 Ran guanine nucleotide release factor_D27K_mutation Human genes 0.000 claims description 3
- 239000012669 liquid formulation Substances 0.000 claims description 3
- 102220095262 rs876659498 Human genes 0.000 claims description 3
- 230000001276 controlling effect Effects 0.000 claims description 2
- 230000001915 proofreading effect Effects 0.000 claims description 2
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 102220244383 rs6500495 Human genes 0.000 claims 1
- 239000004367 Lipase Substances 0.000 abstract description 46
- 108090001060 Lipase Proteins 0.000 abstract description 45
- 102000004882 Lipase Human genes 0.000 abstract description 45
- 235000019421 lipase Nutrition 0.000 abstract description 45
- 230000000694 effects Effects 0.000 abstract description 6
- 230000035772 mutation Effects 0.000 abstract 1
- 235000021391 short chain fatty acids Nutrition 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 21
- 125000003275 alpha amino acid group Chemical group 0.000 description 20
- 229940024606 amino acid Drugs 0.000 description 17
- 235000001014 amino acid Nutrition 0.000 description 17
- 102000004190 Enzymes Human genes 0.000 description 14
- 108090000790 Enzymes Proteins 0.000 description 14
- 238000012360 testing method Methods 0.000 description 13
- 239000004753 textile Substances 0.000 description 13
- 241000228212 Aspergillus Species 0.000 description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 9
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 8
- UYXTWWCETRIEDR-UHFFFAOYSA-N Tributyrin Chemical compound CCCC(=O)OCC(OC(=O)CCC)COC(=O)CCC UYXTWWCETRIEDR-UHFFFAOYSA-N 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 241000223258 Thermomyces lanuginosus Species 0.000 description 7
- 238000004817 gas chromatography Methods 0.000 description 7
- 241000228245 Aspergillus niger Species 0.000 description 6
- 239000002773 nucleotide Substances 0.000 description 6
- 125000003729 nucleotide group Chemical group 0.000 description 6
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 5
- 241000223218 Fusarium Species 0.000 description 5
- 241000235395 Mucor Species 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 239000004744 fabric Substances 0.000 description 5
- 240000006439 Aspergillus oryzae Species 0.000 description 4
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 4
- 229920000742 Cotton Polymers 0.000 description 4
- 241000228143 Penicillium Species 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000006071 cream Substances 0.000 description 4
- 239000003599 detergent Substances 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 241000221779 Fusarium sambucinum Species 0.000 description 3
- 241000144128 Lichtheimia corymbifera Species 0.000 description 3
- 241000223259 Trichoderma Species 0.000 description 3
- 238000002470 solid-phase micro-extraction Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- 241000228215 Aspergillus aculeatus Species 0.000 description 2
- 241001513093 Aspergillus awamori Species 0.000 description 2
- 241001225321 Aspergillus fumigatus Species 0.000 description 2
- 241001480052 Aspergillus japonicus Species 0.000 description 2
- 241000351920 Aspergillus nidulans Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000223651 Aureobasidium Species 0.000 description 2
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241001337994 Cryptococcus <scale insect> Species 0.000 description 2
- 244000163122 Curcuma domestica Species 0.000 description 2
- 235000003392 Curcuma domestica Nutrition 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241000567163 Fusarium cerealis Species 0.000 description 2
- 241000223195 Fusarium graminearum Species 0.000 description 2
- 241000146406 Fusarium heterosporum Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000223198 Humicola Species 0.000 description 2
- 241000235649 Kluyveromyces Species 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-Glutamic acid Natural products OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 125000003338 L-glutaminyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C([H])([H])C(=O)N([H])[H] 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 241000226677 Myceliophthora Species 0.000 description 2
- 241000221960 Neurospora Species 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 241001236817 Paecilomyces <Clavicipitaceae> Species 0.000 description 2
- 241000235648 Pichia Species 0.000 description 2
- 240000005384 Rhizopus oryzae Species 0.000 description 2
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 2
- 235000003534 Saccharomyces carlsbergensis Nutrition 0.000 description 2
- 235000001006 Saccharomyces cerevisiae var diastaticus Nutrition 0.000 description 2
- 244000206963 Saccharomyces cerevisiae var. diastaticus Species 0.000 description 2
- 241001123227 Saccharomyces pastorianus Species 0.000 description 2
- 241000222480 Schizophyllum Species 0.000 description 2
- 241000235346 Schizosaccharomyces Species 0.000 description 2
- 241000228341 Talaromyces Species 0.000 description 2
- 241001540751 Talaromyces ruber Species 0.000 description 2
- 241001494489 Thielavia Species 0.000 description 2
- 241000499912 Trichoderma reesei Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229940091771 aspergillus fumigatus Drugs 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000004422 calculation algorithm Methods 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 235000003373 curcuma longa Nutrition 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 230000008034 disappearance Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000006052 feed supplement Substances 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 229960002989 glutamic acid Drugs 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Natural products C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- -1 methane amide Chemical class 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000013976 turmeric Nutrition 0.000 description 2
- 239000002023 wood Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 241000235389 Absidia Species 0.000 description 1
- 241001375492 Absidia reflexa Species 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 240000000073 Achillea millefolium Species 0.000 description 1
- 235000007754 Achillea millefolium Nutrition 0.000 description 1
- 241001019659 Acremonium <Plectosphaerellaceae> Species 0.000 description 1
- 241000220433 Albizia Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 241000235349 Ascomycota Species 0.000 description 1
- 241000892910 Aspergillus foetidus Species 0.000 description 1
- 241000228232 Aspergillus tubingensis Species 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 241000754798 Calophyllum brasiliense Species 0.000 description 1
- 241000235457 Chytridium Species 0.000 description 1
- 241001362614 Crassa Species 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 241001063191 Elops affinis Species 0.000 description 1
- 235000002756 Erythrina berteroana Nutrition 0.000 description 1
- 241000221207 Filobasidium Species 0.000 description 1
- 241000145614 Fusarium bactridioides Species 0.000 description 1
- 241000223194 Fusarium culmorum Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- 241001112697 Fusarium reticulatum Species 0.000 description 1
- 241001014439 Fusarium sarcochroum Species 0.000 description 1
- 241000223192 Fusarium sporotrichioides Species 0.000 description 1
- 241001465753 Fusarium torulosum Species 0.000 description 1
- 241000567178 Fusarium venenatum Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241001480714 Humicola insolens Species 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241000235087 Lachancea kluyveri Species 0.000 description 1
- 241001344133 Magnaporthe Species 0.000 description 1
- 241001344131 Magnaporthe grisea Species 0.000 description 1
- XUYPXLNMDZIRQH-LURJTMIESA-N N-acetyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC(C)=O XUYPXLNMDZIRQH-LURJTMIESA-N 0.000 description 1
- 241000233892 Neocallimastix Species 0.000 description 1
- 241000221961 Neurospora crassa Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000235379 Piromyces Species 0.000 description 1
- 241000235403 Rhizomucor miehei Species 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 241000204893 Saccharomyces douglasii Species 0.000 description 1
- 241001407717 Saccharomyces norbensis Species 0.000 description 1
- 208000037065 Subacute sclerosing leukoencephalitis Diseases 0.000 description 1
- 206010042297 Subacute sclerosing panencephalitis Diseases 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 241000228178 Thermoascus Species 0.000 description 1
- 241000223257 Thermomyces Species 0.000 description 1
- 241001313536 Thermothelomyces thermophila Species 0.000 description 1
- 241001149964 Tolypocladium Species 0.000 description 1
- 241000223260 Trichoderma harzianum Species 0.000 description 1
- 241000378866 Trichoderma koningii Species 0.000 description 1
- 241000223262 Trichoderma longibrachiatum Species 0.000 description 1
- 241000223261 Trichoderma viride Species 0.000 description 1
- 241000235013 Yarrowia Species 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N aspartic acid group Chemical group N[C@@H](CC(=O)O)C(=O)O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000009954 braiding Methods 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- HKPHPIREJKHECO-UHFFFAOYSA-N butachlor Chemical compound CCCCOCN(C(=O)CCl)C1=C(CC)C=CC=C1CC HKPHPIREJKHECO-UHFFFAOYSA-N 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 230000002366 lipolytic effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000015927 pasta Nutrition 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 229910021654 trace metal Inorganic materials 0.000 description 1
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 230000004304 visual acuity Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
Abstract
本发明提供通过将突变引入在亲本脂肪酶中鉴定的一个或多个区域而获取的多肽。令人惊奇的发现本发明的多肽针对短链脂肪酸具有低比活性，导致与本领域已知的脂肪酶相比减少的气味产生和增加的BR。
Description
发明领域
本发明涉及相对气味产生具有改进的洗涤效果的脂肪酶变体，及制备它们的方法。其特别涉及细毛嗜热霉(Thermomyces lanuginosus)脂肪酶的变体。
背景技术
脂肪酶是有用的，例如，作为洗涤剂酶用于从衣物和其它纺织品去除脂质或脂肪污渍，及作为用于面包和其它烘焙产品的生面团的添加剂。因此，源自细毛嗜热霉(Thermomyces lanuginosus)(同物异名疏棉状腐质霉(Humicola lanuginosa)，EP 258068和EP 305216)的脂肪酶以商品名
(Novozymes A/S的产品)出售用于洗涤剂用途。WO 0060063描述了细毛嗜热霉脂肪酶的变体，其在洗涤剂溶液中具有特别好的初次洗涤性能(first-wash performance)。WO 9704079、WO 9707202和WO 0032758也公开了细毛嗜热霉脂肪酶的变体。
在一些应用中，感兴趣的是将产生气味的短链脂肪酸的形成降至最低。因此，已知的是含有脂肪酶的洗衣洗涤剂有时可能留下残留的气味沾在受到牛奶污染的衣物上(EP 430315)。WO 02062973公开了脂肪酶变体，其中通过附接(attaching)C末端延伸减少了气味的产生。最近公开的WO 07087508公开了脂肪酶变体，其中通过在亲本脂肪酶中鉴定的一个或多个区域中引入突变而减少了气味产生。WO 07087503描述了具有脂肪酶活性的多肽，且其在该说明书中给定的测试条件下进一步具有至少0.8的RP和至少1.1的BR。
发明内容
在第一方面，本发明涉及具有脂肪酶活性的第一多肽，其中所述多肽是具有下列中的至少一项的多肽：(a)相对于280nm处的吸光度(A280)的脂肪酶活性(LU)少于500LU/A280，其中一单位的LU(1LU)定义为能够在30℃，pH7每分钟释放1μmol的丁酸的的酶量，且所述多肽的吸光度在280nm测定；(b)风险性能气味(Risk performance odor，R)低于0.5，其中R计算为自多肽洗涤样本释放的丁酸量和自参照多肽洗涤样本释放的丁酸量的比例，计算比例之前两个值均相对于自非多肽洗涤样本释放的丁酸量进行了校正；或者(c)效益风险因子(Benefit Risk factor，BR)至少为1.8，其中BR定义为平均洗涤性能(average wash performance，RPavg)除以风险性能气味(R)。
在第二方面，本发明涉及具有脂肪酶活性的第二多肽，其包含在下述位置的氨基酸的改变：T231R+N233R+I255A+P256K及下列中的至少一种：(a)S58A+V60S+A150G+L227G；或(b)E210V/G，所述位置对应于SEQ ID NO：2。
在进一步的方面，本发明涉及编码具有脂肪酶活性的多肽的分离的多核苷酸、包含该多核苷酸的核酸构建体、包含所述核酸构建体的重组表达载体以及包含所述核酸构建体或所述重组表达载体的转化的宿主细胞。
在进一步的方面，本发明涉及制备所述多肽的方法，包括下述步骤：(a)在有益于所述多肽产生的条件下培养包含所述核酸构建体或重组表达载体的转化宿主细胞，所述核酸构建体或重组表达载体包含所述多肽；以及(b)回收该多肽。
在进一步的方面，本发明涉及包含所述多肽的配制物。
在进一步的方面，本发明涉及通过使用所述多肽在脂质水解过程中减少产生气味的短链脂肪酸形成的方法。
附图简述
图1显示脂肪酶的比对。
序列列表
SEQ ID NO：1显示编码来自细毛嗜热霉的脂肪酶的DNA序列。
SEQ ID NO：2显示来自细毛嗜热霉的脂肪酶的氨基酸序列。
SEQ ID NO：3显示来自反射犁头霉(Absidia reflexa)的脂肪酶的氨基酸序列。
SEQ ID NO：4显示来自伞枝犁头霉(Absidia corymbifera)的脂肪酶的氨基酸序列。
SEQ ID NO：5显示来自曼赫根毛霉(Rhizmucor miehei)的脂肪酶的氨基酸序列。
SEQ ID NO：6显示来自米根霉(Rhizopus oryzae)的脂肪酶的氨基酸序列。
SEQ ID NO：7显示来自黑曲霉(Aspergillus niger)的脂肪酶的氨基酸序列。
SEQ ID NO：8显示来自塔宾曲霉(Aspergillus tubigensis)的脂肪酶的氨基酸序列。
SEQ ID NO：9显示来自尖镰孢(Fusarium oxysporum)的脂肪酶的氨基酸序列。
SEQ ID NO：10显示来自异孢镰孢(Fusarium heterosporum)的脂肪酶的氨基酸序列。
SEQ ID NO：11显示来自米曲霉(Aspergillus oryzea)的脂肪酶的氨基酸序列。
SEQ ID NO：12显示来自沙门柏干酪青霉(Penicilium camembertii)的脂肪酶的氨基酸序列。
SEQ ID NO：13显示来自臭曲霉(Aspergillus foetidus)的脂肪酶的氨基酸序列。
SEQ ID NO：14显示来自黑曲霉的脂肪酶的氨基酸序列。
SEQ ID NO：15显示来自米曲霉的脂肪酶的氨基酸序列。
SEQ ID NO：16显示来自Landerina penisapora的脂肪酶的氨基酸序列。
发明详述
使用脂肪酶去除脂质和脂肪污渍(fatty stain)在本领域是已知的，其中导致游离短链脂质(如丁酸)的释放的脂肪酶活性与令人不快的(undesirable)气味相关。底物三丁酸甘油酯的水解导致丁酸的释放。令人惊讶地发现本发明的多肽在中性pH针对三丁酸甘油酯具有低比活性(作为LU/A280测定)，参见实施例2和表3。
效益风险因子(BR)通过相对(洗涤)性能(效益，RP)除以风险性能气味(风险，R)来计算。所述洗涤性能可通过自动机械应力试验(automated mechanical stress assay，AMSA)来测定，参见实施例3，而气味产生可直接通过气相层析来测定，参见实施例4和表3。气味减少影响BR，且可导致BR增加。另外还发现本发明的多肽与本领域已知的脂肪酶相比较具有减少的气味产生和增加的BR，参见实施例5和表3。
脂肪酶活性(LU)：如本文所使用的术语“脂肪酶活性”指催化在二酰基甘油和羧酸的形成下三酰基甘油的水解的羧酸酯水解酶活性。就本发明而言，脂肪酶活性通过下述过程来确定：脂肪酶的底物通过使用阿拉伯胶作为乳化剂对三丁酸甘油酯(甘油三丁酸)进行乳化来制备。接着在30℃，pH 7或9在pH恒定的滴定实验中水解三丁酸甘油酯。一个单位的脂肪酶活性(1LU)定义为能够在30℃，pH7每分钟释放1μmol丁酸的酶量。
风险性能气味(R)：如本文所使用的术语“风险性能气味”指自多肽洗涤样本释放的丁酸量和自参照多肽洗涤样本释放的丁酸量之间的比例，之前两个值均相对于自非多肽洗涤样本释放的丁酸量进行了校正。
相对性能(RP)：如本文所使用的术语“相对性能”指与参照多肽的洗涤性能相比较的所述多肽的洗涤性能。就本发明而言，相对性能根据在实施例3中所描述的过程来确定。
参照多肽：如本文所使用的术语“参照多肽”、“参照酶”或“参照脂肪酶”指具有取代T231R+N233R的SEQ ID NO：2的成熟部分。
效益风险因子(BR)：如本文所使用的术语“效益风险因子”指与气味产生风险(R)相比较的平均相对性能(RPavg)，且具有下列公式：BR＝RPavg/R。
氨基酸修饰的命名法
为了易于参考，在描述本发明的脂肪酶变体时，采用了以下命名法：
原始氨基酸：位置：取代氨基酸
根据这种命名法，例如，在位置195中用谷氨酸取代甘氨酸表示为G195E。在相同的位置中缺失甘氨酸表示为G195*，而插入额外的氨基酸残基例如赖氨酸表示为G195GK。与其它脂肪酶相比，当特定脂肪酶含有“缺失”并且在该位置进行插入的情况，就在位置36插入天冬氨酸而言，将这种情况表示为*36D。
用加号分隔多个突变，即：R170Y+G195E，分别表示在位置170用酪氨酸取代精氨酸，和在位置195用谷氨酸取代甘氨酸。
当应用所述排列方法时，X231表示在亲本多肽中对应于位置231的氨基酸。X231R表示用R置换所述氨基酸。就SEQ ID NO：2而言，X是T，因此X231R表示用R取代位置231的T。当某个位置(例如231)中的氨基酸可以由选自一组氨基酸的另一个氨基酸取代的情况，例如由R和P和Y组成的组，将这种情况表示为X231R/P/Y。
在所有的情况下，采用公认的IUPAC单字母或三字母的氨基酸缩写。
同一性：如本文所使用的术语“同一性”指两个氨基酸序列之间或两个核苷酸序列之间的相关性，由参数“同一性”来描述。
就本发明而言，通过使用来自EMBOSS软件包(http://emboss.org)版本2.8.0的Needle程序来测定两个氨基酸序列之间的比对。Needle程序执行总体比对算法(global alignment algorithm)，其在Needleman，S.B.and Wunsch，C.D.(1970)J.Mol.Biol.48，443-453中描述。使用的取代矩阵是BLOSUM62，缺口开启罚分(gap opening penalty)是10，并且缺口延伸罚分(gap extension penalty)是0.5。
本发明的氨基酸序列(“发明序列”；例如SEQ ID NO：2的氨基酸1至269)和不同氨基酸序列(“外源序列”)之间的同一性程度如下计算：用两个序列比对中的精确匹配数除以“发明序列”的长度或“外源序列”的长度中最短的一个。将结果表示为百分比同一性。
当“发明序列”和“外源序列”在重叠的相同位置中具有相同的氨基酸残基时，发生精确匹配。序列的长度是序列中氨基酸残基的数目(例如SEQ ID NO：2的长度是269)。
可以使用上述方法计算同一性和同源性，并用于比对。在本发明的上下文中，同源性和比对如下所述计算。
同源性和比对
就本发明而言，可以通过本领域已知的计算机程序的手段合适地测定同源性的程度，所述程序例如GCG程序包中提供的GAP(Program Manual for the Wisconsin Package，Version 8，August 1994，Genetics Computer Group，575Science Drive，Madison，Wisconsin，USA 53711)(Needleman，S.B.and Wunsch，C.D.，(1970)，Journal of Molecular Biology，48，443-45)，按照用于多肽序列比较的以下设定来使用GAP：GAP产生罚分(GAP creation penalty)为3.0，和GAP延伸罚分(GAP extension penalty)为0.1。
在本发明中，通过图1所示的比对定义反射犁头霉、伞枝犁头霉(Absidia corymbefera)、曼赫根毛霉、德氏根霉、黑曲霉、塔宾曲霉、尖镰孢、异孢镰孢、米曲霉、沙门柏干酪青霉、臭曲霉、黑曲霉、细毛嗜热霉(同物异名：疏棉状腐质霉)和Landerina penisapora的脂肪酶序列中相应(或同源)的位置。
为了发现所述比对中未显示的脂肪酶序列中的同源位置，将感兴趣的序列与图1中显示的序列进行比对。通过使用GAP比对将新序列与GAP程序发现的最同源的序列进行比对，将所述新序列排入图1中的现有比对。GAP在GCG程序包(Program Manual for the Wisconsin Package，Version 8，August1994，Genetics Computer Group，575 Science Drive，Madison，Wisconsin，USA53711)(Needleman，S.B.and Wunsch，C.D.，(1970)，Journal ofMolecular Biology，48，443-45)中提供。使用以下设置用于多肽序列比较：GAP产生罚分为3.0，且GAP延伸罚分为0.1。
具有脂肪酶活性多肽的来源
可以使用任何合适的多肽。在一些实施方案中，所述多肽可以是真菌多肽。
所述多肽可为源自下列属的酵母多肽：如，念珠菌属(Candida)、克鲁维酵母属(Kluyveromyces)、毕赤酵母属(Pichia)、酵母属(Saccharomyces)、裂殖酵母属(Schizosaccharomyces)或西洋蓍霉属(Yarrowia)多肽；或更优选是源自下列属的丝状真菌多肽：如，枝顶孢霉属(Acremonium)、曲霉属(Aspergillus)、短梗霉属(Aureobasidium)、隐球菌属(Cryptococcus)、Filobasidium、镰孢属(Fusarium)、腐质霉属(Humicola)、梨孢菌属(Magnaporthe)、毛霉属(Mucor)、毁丝霉属(Myceliophthora)、新考玛脂霉属(Neocallimastix)、脉孢菌属(Neurospora)、拟青霉属(Paecilomyces)、青霉属(Penicillium)、瘤胃壶菌属(Piromyces)、裂褶菌属(Schizophyllum)、踝节菌属(Talaromyces)、嗜热子囊菌属(Thermoascus)、梭孢壳属(Thielavia)、弯颈霉属(Tolypocladium)、嗜热霉属(Thermomyces)或木霉属(Trichoderma)。
另外，所述多肽可为具有脂肪酶活性的卡尔酵母(Saccharomyces carlsbergensis)、酿酒酵母(Saccharomyces cerevisiae)、糖化酵母(Saccharomyces diastaticus)、道格拉氏酵母(Saccharomyces douglasii)、克鲁弗酵母(Saccharomyces kluyveri)、诺地酵母(Saccharomyces norbensis)或卵形酵母(Saccharomyces oviformis)多肽。
或者，所述多肽是棘孢曲霉(Aspergillus aculeatus)、泡盛曲霉(Aspergillus awamori)、烟曲霉(Aspergillus fumigatus)、臭曲霉、日本曲霉(Aspergillus japonicus)、构巢曲霉(Aspergillus nidulans)、黑曲霉、米曲霉、塔宾曲霉、杆孢状镰孢(Fusarium bactridioides)、禾谷镰孢(Fusarium cerealis)、库威镰孢(Fusarium crookwellense)、大刀镰孢(Fusarium culmorum)、禾本科镰孢(Fusarium graminearum)、禾赤镰孢(Fusarium graminum)、异孢镰孢、合欢木镰孢(Fusarium negundi)、尖镰孢、多枝镰孢(Fusarium reticulatum)、粉红镰孢(Fusarium roseum)、接骨木镰孢(Fusarium sambucinum)、肤色镰孢(Fusariumsarcochroum)、拟分枝孢镰孢(Fusarium sporotrichioides)、硫色镰孢(Fusariumsulphureum)、圆镰孢(Fusarium torulosum)、拟丝孢镰孢(Fusarium trichothecioides)、镶片镰孢(Fusarium venenatum)、特异腐质霉(Humicolainsolens)、细毛嗜热霉(同物异名：疏棉状腐质霉)、米赫毛霉(Mucor miehei)、嗜热毁丝霉(Myceliophthora thermophila)、粗糙脉孢菌(Neurospora crassa)、产紫青霉(Penicillium purpurogenum)、哈茨木霉(Trichoderma harzianum)、康宁木霉(Trichoderma koningii)、长枝木霉(Trichoderma longibrachiatum)、里氏木霉(Trichoderma reesei)或绿色木霉(Trichoderma viride)多肽。
在一些实施方案中，本发明涉及是嗜热霉属脂肪酶的多肽。
在一些实施方案中，本发明涉及是细毛嗜热霉脂肪酶的多肽。
在一些实施方案中，本发明涉及下述多肽，其中所述多肽与SEQ ID NO：2至少50％，至少60％，至少70％，至少80％，至少85％，至少90％，至少95％，至少96％，至少97％，至少98％，至少99％或100％相同。
鉴定具有脂肪酶活性的多肽中的改变
下面提及的位置是SEQ ID NO：2中氨基酸残基的位置。将在“同源性和比对”段落所描述的过程用于在不同脂肪酶中寻找氨基酸残基的对应或同源位置。
在一些实施方案中，本发明涉及具有脂肪酶活性的第一多肽，其中所述多肽是具有下列中的至少一项的多肽：(a)相对于280nm处的吸光度(A280)的脂肪酶活性(LU)少于500，少于450，少于400，少于350，少于300，少于250，少于200，少于150，少于150或少于50LU/A280，其中一单位的LU(1LU)定义为能够在30℃，pH 7每分钟释放1μmol的丁酸的酶量，且所述多肽的吸光度在280nm测定；(b)风险性能气味(Risk performance odor，R)低于0.5，低于0.4，低于0.3，低于0.2，低于0.1或低于0.05，其中将R计算为自多肽洗涤样本释放的丁酸量和自参照多肽洗涤样本释放的丁酸量之间的比例，计算比例之前两个值均相对于自非多肽洗涤样本释放的丁酸量进行了校正；或(c)效益风险因子(Benefit Risk factor，BR)为至少1.8，至少1.9，至少2.0，至少2.5，至少3.0，至少4.0，至少5.0或至少6.0，其中BR定义为平均洗涤性能(average wash performance，RPavg)除以风险性能气味(R)。
在一些实施方案中，本发明涉及第一多肽，其中所述多肽包含在下述位置的氨基酸的改变：T231R+N233R+I255A+P256K和下列中的至少一项：(a)S58A+V60S+A150G+L227G；或(b)E210V/G；所述位置对应于SEQ ID NO：2。
在一些实施方案中，本发明涉及进一步包含在位置I86V或T143S的氨基酸改变中的至少一种的第一多肽。
在一些实施方案中，本发明涉及第一多肽，其中所述多肽包含至少一个选自下列的进一步改变：在对应于SEQ ID NO：2的位置E1、D27、N33、S83、G91、N94、K98、E99、D102、D111、G163、I202、E210、S216、L259或L269的位置取代、缺失或添加至少一个氨基酸。
在一些实施方案中，本发明涉及第一多肽，其中所述至少一个改变选自下组：SEQ ID NO：2的E1N/*、D27N、N33Q、S83T、G91N、N94R、K98I、E99K、D102A、D111N、G163K、I202L、E210A、S216P、L259F或L269APIA。
在一些实施方案中，本发明涉及第二多肽，其包含在下列位置的氨基酸的改变：T231R+N233R+I255A+P256K和下列中的至少一种：(a)S58A+V60S+A150G+L227G；或(b)E210V/G；所述位置对应于SEQ ID NO：2。
在一些实施方案中，本发明涉及进一步包含在位置I86V或T143S的氨基酸改变中的至少一种的第二多肽。
在一些实施方案中，本发明涉及第二多肽，其中所述多肽包含至少一个选自下列的进一步改变：在对应于SEQ ID NO：2的位置E1、D27、N33、S83、G91、N94、K98、E99、D102、D111、G163、I202、E210、S216、L259或L269位置的位置取代、缺失或添加至少一个氨基酸。
在一些实施方案中，本发明涉及第二多肽，其中所述至少一个改变选自下组：SEQ ID NO：2的E1N/*、D27N、N33Q、S83T、G91N、N94R、K98I、E99K、D102A、D111N、G163K、I202L、E210A、S216P、L259F或L269APIA。
在一些实施方案中，本发明涉及第一多肽，其中所述多肽包含选自下组的改变：(a)T231R+N233R+L269APIA；(b)S58T+V60K+A150G+T231R+N233I+D234G；(c)S58T+V60K+I86V+D102A+A150G+L227G+T231R+N233R+P256K；(d)S58N+V60S+I86P+T231R+N233R+P256S；(e)S58N+V60S+I86S+L227G+T231R+N233R+P256S；以及(f)S58N+V60S+I86T+L227G+T231R+N233R+P256L。
在一些实施方案中，本发明涉及第一或第二多肽，其中所述多肽包含选自下组的改变：(a)S58A+V60S+S83T+A150G+L227G+T231R+N233R+I255A+P256K；(b)S58A+V60S+I86V+A150G+L227G+T231R+N233R+I255A+P256K；(c)S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(d)S58A+V60S+I86V+T143S+A150G+G163K+S216P+L227G+T231R+N233R+I255A+P256K；(e)E1*+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(f)S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(g)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K+L259F；(h)S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(i)N33Q+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(j)E1*+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(k)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；(l)D27N+S58A+V60S+I86V+G91N+N94R+D111N+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(m)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+E210A+S216P+L227G+T231R+N233R+I255A+P256K；(n)A150G+E210V+T231R+N233R+I255A+P256K；(o)I202L+E210G+T231R+N233R+I255A+P256K；(p)E1N+A18K+V60K+I86V+A150G+E210A+L227G+T231R+N233R+P256K；(q)E1L+D27K+V60K+I86V+A150G+S219P+L227G+T231R+N233R+P256K；(r)E1N+S58A+V60S+S83T+A150G+L227G+T231R+N233R+I255A+P256K；(s)E1N+S58T+V60K+I86V+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(t)E1N+S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；和(u)S58A+V60S+S83T+A150A+L227G+T231R+N233R+I255A+P256K。
表1：在所述多肽中可能包含的改变
在一些实施方案中，本发明涉及第一多肽，其中所述多肽包含选自下组的改变：(a)T231R+N233R+L269APIA；(b)S58T+V60K+A150G+T231R+N233I+D234G；(c)S58T+V60K+I86V+D102A+A150G+L227G+T231R+N233R+P256K；(d)S58N+V60S+I86P+T231R+N233R+P256S；(e)S58N+V60S+I86S+L227G+T231R+N233R+P256S；和(f)S58N+V60S+I86T+L227G+T231R+N233R+P256L。
在一些实施方案中，本发明涉及第一或第二多肽，其中所述多肽包含选自下组的改变：(a)S58A+V60S+S83T+A150G+L227G+T231R+N233R+I255A+P256K；(b)S58A+V60S+I86V+A150G+L227G+T231R+N233R+I255A+P256K；(c)S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(d)S58A+V60S+I86V+T143S+A150G+G163K+S216P+L227G+T231R+N233R+I255A+P256K；(e)E1*+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(f)S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(g)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K+L259F；(h)S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(i)N33Q+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(j)E1*+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(k)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；(l)D27N+S58A+V60S+I86V+G91N+N94R+D111N+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(m)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+E210A+S216P+L227G+T231R+N233R+I255A+P256K；(n)A150G+E210V+T231R+N233R+I255A+P256K；(o)I202L+E210G+T231R+N233R+I255A+P256K；(p)E1N+A18K+V60K+I86V+A150G+E210A+L227G+T231R+N233R+P256K；(q)E1L+D27K+V60K+I86V+A150G+S219P+L227G+T231R+N233R+P256K；(r)E1N+S58A+V60S +S83T+A150G+L227G+T231R+N233R+I255A+P256K；(s)E1N+S58T+V60K+I86V+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；(t)E1N+S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；和(u)S58A+V60S+S83T+A150A+L227G+T231R+N233R+I255A+P256K。
多肽的多核苷酸、表达载体、宿主细胞、产生
在一些实施方案中，本发明涉及编码所述多肽的分离的多核苷酸。这些多核苷酸在非常低严紧性条件下，优选低严紧性条件下，更优选中严紧性条件下，更优选中-高严紧性条件下，甚至更优选高严紧性条件下，并且最优选非常高严紧性条件下，可以与以下杂交：(i)SEQ ID NO：1的核苷酸178至660，(ii)SEQ ID NO：1的核苷酸178至660中包含的cDNA序列，(iii)(i)或(ii)的亚序列，或(iv)(i)、(ii)或(iii)的互补链(J.Sambrook，E.F.Fritsch，and T.Maniatus，1989，Molecular Cloning，A Laboratory Manual，2d edition，Cold Spring Harbor，New York)。SEQ ID NO：1的亚序列含有至少100个连续的核苷酸或优选至少200个连续的核苷酸。此外，所述亚序列可编码具有脂肪酶活性的多肽片段。
对于长度至少100个核苷酸的长探针，将非常低至非常高的严紧条件定义为：于42℃在5×SSPE、0.3％SDS、200μg/ml剪切和变性的鲑精DNA，以及对于非常低和低严紧性25％的甲酰胺，对于中和中-高严紧性35％的甲酰胺，或者对于高和非常高严紧性50％的甲酰胺中，根据标准Southern印迹方法最佳进行12至24小时的预杂交和杂交。
对于长度至少为100个核苷酸的长探针，使用2×SSC、0.2％SDS，优选至少在45℃(非常低严紧性)，更优选至少50℃(低严紧性)，更优选至少55℃(中等严紧性)，更优选至少60℃(中-高严紧性)，甚至更优选至少65℃(高严紧性)，和最优选至少70℃(非常高严紧性)将载体材料最终清洗三次，每次15分钟。
在一些实施方案中，本发明涉及包含所述多核苷酸的核酸构建体，所述多核苷酸可操作连接于至少一个在表达宿主中指导所述多肽产生的调控序列。
在一些实施方案中，本发明涉及包含所述核酸构建体的重组表达载体。
在一些实施方案中，本发明涉及包含所述核酸构建体或所述重组表达载体的转化的宿主细胞。
编码所述多核苷酸的分离的多核苷酸、包含所述多核苷酸的核酸构建体、包含所述核酸构建体的重组表达载体以及包含所述核酸构建体或所述重组表达载体的转化的宿主细胞均可通过本领域已知的方法获取。
在一些实施方案中，本发明涉及制备多肽的方法，包括下述步骤：(a)在有益于所述多肽生产的条件下培养所述转化的宿主细胞，所述宿主细胞包含所述核酸构建体或包含所述核酸构建体的重组表达载体；和(b)回收该多肽。本方法可根据本领域已知的原则加以实施。
用途
本发明的酶可用于，包括供去除脂肪物质用的工业用途。
在一些实施方案中，本发明涉及包含所述多肽的配制物。在进一步的实施方案中，本发明涉及配制物，其中所述配制物可为固体或液体配制物。所述多肽可用于固体以及液体配制物两者。
在一些实施方案中，本发明涉及通过使用所述多肽在脂质水解过程中减少产生气味的短链脂肪酸形成的方法。
实施例
本发明进一步通过下述实施例进行描述，其不应理解为对本发明范围的限制。
用作缓冲剂和底物的化学品是至少试剂等级的商业产品。
实施例1-脂肪酶变体的产生
使用本领域的标准方法构建含有编码所述多肽的基因的质粒，并且将所述质粒转化至合适的宿主细胞中。
使用34℃的恒定培养基温度和1.2升的起始体积，按照补料分批发酵来进行发酵。将培养基的初始pH设为6.5。一旦pH增加至7.0，则通过添加10％H3PO4来保持该值。通过改变搅拌速率和使用1.0升空气每升培养基每分钟的固定通气速率来控制培养基中的溶解氧水平。在整个补料分批阶段期间，将补料添加速率保持在恒定水平。
分批培养基含有麦芽糖糖浆作为碳源，尿素和酵母提取物作为氮源，以及痕量金属和盐的混合物。在补料分批阶段期间连续添加的补料含有麦芽糖糖浆作为碳源，而添加酵母提取物和尿素以确保氮的充足供应。
可以通过使用本领域已知的标准方法来进行所述多肽的纯化，例如通过过滤发酵上清，继之以疏水层析和离子交换层析，例如如EP 0 851 913 EP，实施例3中所述。
实施例2-相对于280nm处吸光度的脂肪酶活性单位(LU)(LU/A280)
脂肪酶活性(LU)如上面在脂肪酶活性部分所述来确定。测定所述脂肪酶在280nm的吸光度(A280)。多肽的比活性可表达为LU/A280的比例。
相对LU/A280由所述多肽的LU/A280除以参照酶的LU/A280来计算。在本发明的背景下，所述参照酶是具有取代T231R+N223R的SEQ ID NO：2的脂肪酶。
实施例3-由自动机械应力试验(AMSA)获取的数据计算相对性能(RP)
使用自动机械应力试验(Automatic Mechanical Stress Assay；AMSA)测试本发明的多肽。使用AMSA测试，能够检查大量小体积酶洗涤剂溶液的洗涤性能。AMSA平板具有用于测试溶液的多个槽(slot)，和将待洗涤的纺织品样本相对于所有槽的开口(slot opening)压紧的盖。在洗涤时间中，剧烈摇动平板、测试溶液、纺织品和盖以使测试溶液与纺织品接触并且施以机械应力。进一步的描述参见WO 02/42740，特别是第23-24页的段落“特殊方法实施方式”。含有洗涤剂测试溶液的容器由金属平板中的圆柱形孔洞(直径6mm，深10mm)组成。沾污的织物(测试材料)放置在金属平板顶部，并将沾污的织物用作盖子密封在所述容器上。另一个金属平板放置在沾污织物的顶部以避免从各个容器中的任何溢出。以2mm振幅和30Hz的频率将两块金属平板连同沾污织物一起上下振动。
表2：AMSA的实验条件
通过将5g姜黄(Santa Maria，Denmark)与100g奶油(38％脂肪，Arla，Denmark)和100克咖啡稀奶油(coffee cream)(9％脂肪Arla，Denmark)在50℃混合来分别制备奶油-姜黄样本和咖啡稀奶油姜黄样本。将所述混合物在该温度保持大约20分钟，并且过滤(50℃)以去除任何未溶解的颗粒。将混合物冷却至20℃，并且将编织的棉布样本EMPA221浸入所述奶油-姜黄混合物，之后允许其在室温干燥过夜，并且冷冻直到使用。奶油-姜黄样本的制备在WO06125437中公开。
将多肽的性能作为用所述特定多肽洗涤的纺织品样品的颜色亮度来测量。也可以将亮度表示为当用白光照射时，从纺织品样品反射的光的强度。当纺织品被沾污时，反射光的强度与清洁纺织品的反射光强度相比较低。因此，能够使用反射光的强度来测量多肽变体的洗涤性能。
使用专业平板扫描仪(PFU DL2400pro)进行颜色测量，使用该扫描仪来捕捉经洗涤纺织品样品的图像。使用200dpi的分辨率和24比特(bit)的输出色深度来进行扫描。为了获得精确的结果，经常性地使用Kodak反射IT8指标(Kodak reflective IT8 target)校准扫描仪。
为了从扫描的图像提取光强度的值，使用特殊设计的软件应用程序(Novozymes Color Vector Analyzer)。该程序从所述图像恢复24比特像素值，并且将它们转化为红、绿和蓝的值(RGB)。通过将RGB值作为矢量加在一起，再取所得矢量的长度来计算强度值(Int)：
根据下式计算多肽的洗涤性能(P)：
P＝Int(v)-Int(r)，
其中Int(v)是用酶洗涤的纺织品表面的光强度值，且Int(r)是不用酶洗涤的纺织品表面的光强度值。
根据以下定义，给出相对性能评分作为AMSA洗涤的结果：
相对性能评分(RP)概括了测试多肽相对于参照多肽的性能(P)：
RP＝P(测试多肽)/P(参照多肽)。
RPavg表示在0.5mg ep/1处进行的测定中与参照多肽比较的平均相对性能。
如果多肽表现优于参照，则认为多肽呈现出改进的洗涤性能。在本发明的上下文中，参照酶是具有取代T231R+N233R的SEQ ID NO：2的脂肪酶。
实施例4-由固相微抽提气相层析(Solid Phase Micro Extraction Gas Chromatograph)测定计算风险因子(R)
使用以下方法通过固相微抽提气相层析(Solid Phase Micro Extraction Gas Chromatography)(SPME-GC)测量来自经脂肪酶洗涤的样本的丁酸释放。将在含有0.5mg/L脂肪酶的表2中的指定溶液中洗涤的四个纺织品片(直径5mm)转移至气相层析(GC)小瓶。将样品在30℃下温育24h，并随后加热至140℃30分钟，并在20℃-25℃下储藏至少4小时直至进行分析。在装备有Stabilwax-DA w/Integra-Guard柱(30m，0.32mm ID和0.25微米df)和Carboxen PDMSSPME纤维(85微米)的Varian 3800GC上进行分析。对每个GC小瓶的采样在50℃下用SPME纤维在纺织品片之上的顶部空间(head space)中进行8分钟，且继而将所采样的化合物注射到柱上(注射器温度＝250℃)。柱流速＝2ml氦/分钟。柱加热炉温度梯度：0分钟＝50℃，2分钟＝50℃，6分45秒＝240℃，11分45秒＝240℃。使用火焰离子化检测器(Flame Ionization Dector，FID)进行检测，并使用可靠的标准鉴别出丁酸的保留时间。
多肽的风险性能气味(R)指自多肽洗涤样本释放的丁酸量(峰面积)和自参照多肽洗涤样本释放的丁酸量(峰面积)之间的比例，之前两个值均相对于自非多肽洗涤样本(空白)释放的丁酸量(峰面积)进行了校正。所述参照多肽是具有取代T231R+N223R的SEQ ID NO：2的多肽。所述多肽的风险性能气味(R)根据下式计算：
气味＝测得的自纺织物表面释放的丁酸(峰面积)
α测试酶＝气味测试酶-气味空白
α参照酶＝气味参照酶-气味空白
R＝α测试酶/α参照酶
如果R因子小于1，则认为多肽与参照相比表现出减少的气味。
实施例5-效益风险因子(BR)
效益风险因子描述的是与减少的气味风险相比的洗涤性能，因此将其定义为：
BR＝RPavg/R
如果BR因子高于1，则认为变体显示出改进的洗涤性能和减少的气味。
表3：一些本发明多肽的比活性(LU/A280)、风险性能气味(R)和效益风险因子(BR)
Claims (20)
1.一种具有脂肪酶活性的多肽，其中所述多肽是具有下列中的至少一项的多肽：
(a)相对于280nm处的吸光度(A280)的脂肪酶活性(LU)少于500LU/A280，其中一单位的LU(1LU)定义为能够在30℃，pH7每分钟释放1μmol的丁酸的酶量，且所述多肽的吸光度在280nm处测定；
(b)风险性能气味(R)低于0.5，其中R计算为自多肽洗涤样本释放的丁酸量和自参照多肽洗涤样本释放的丁酸量的比例，计算比例之前两个值均相对于自非多肽洗涤样本释放的丁酸量进行了校正；或
(c)效益风险因子(BR)至少为1.8，其中BR定义为平均洗涤性能(RPavg)除以风险性能气味(R)。
2.权利要求1的多肽，其包含在下述位置的氨基酸改变：T231R+N233R+I255A+P256K和下列中的至少一种：
(a)S58A+V60S+A150G+L227G；或
(b)E210V/G；
所述位置对应于SEQ ID NO：2。
3.权利要求2的多肽，进一步在位置I86V或T143S处包含至少一个氨基酸改变。
4.权利要求2-3任一所述的多肽，其中所述多肽包含至少一个选自下列的进一步改变：在对应于SEQ ID NO：2的位置E1、D27、N33、S83、G91、N94、K98、E99、D102、D111、G163、I202、E210、S216、L259或L269的位置取代、缺失或添加至少一个氨基酸。
5.权利要求4的多肽，其中所述至少一个改变选自下组：SEQ ID NO：2的E1N/*、D27N、N33Q、S83T、G91N、N94R、K98I、E99K、D102A、D111N、G163K、I202L、E210A、S216P、L259F或L269APIA。
6.一种多肽，其包含在下述位置的氨基酸改变：T231R+N233R+I255A+P256K和下列中的至少一种：
(a)S58A+V60S+A150G+L227G；或
(b)E210V/G；
其中所述位置对应于SEQ ID NO：2。
7.权利要求6的多肽，其进一步在位置I86V或T143S处包含至少一个氨基酸改变。
8.权利要求6-7任一所述的多肽，其中所述多肽包含至少一个选自下列的进一步改变：在对应于SEQ ID NO：2中位置E1、D27、N33、S83、G91、N94、K98、E99、D102、D111、G163、I202、E210、S216、L259或L269的位置取代、缺失或添加至少一个氨基酸。
9.权利要求8的多肽，其中所述至少一个改变选自下组：SEQ ID NO：2的E1N/*、D27N、N33Q、S83T、G91N、N94R、K98I、E99K、D102A、D111N、G163K、I202L、E210A、S216P、L259F或L269APIA。
10.权利要求1的多肽，其中所述多肽包含选自下组的改变：
(a)T231R+N233R+L269APIA；
(b)S58T+V60K+A150G+T231R+N233I+D234G；
(c)S58T+V60K+I86V+D102A+A150G+L227G+T231R+N233R+P256K；
(d)S58N+V60S+I86P+T231R+N233R+P256S；
(e)S58N+V60S+I86S+L227G+T231R+N233R+P256S；和
(f)S58N+V60S+I86T+L227G+T231R+N233R+P256L。
11.权利要求1或6的多肽，其中所述多肽包含选自下组的改变：
(a)S58A+V60S+I83T+A150G+L227G+T231R+N233R+I255A+P256K；
(b)S58A+V60S+I86V+A150G+L227G+T231R+N233R+I255A+P256K；
(c)S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(d)S58A+V60S+I86V+T143S+A150G+G163K+S216P+L227G+T231R+N233R+I255A+P256K；
(e)E1*+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(f)S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(g)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K+L259F；
(h)S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(i)N33Q+S58A+V60S+I86V+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(j)E1*+S58A+V60S+I86V+K98I+E99K+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(k)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；
(l)D27N+S58A+V60S+I86V+G91N+N94R+D111N+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(m)E1N+S58A+V60S+I86V+K98I+E99K+T143S+A150G+E210A+S216P+L227G+T231R+N233R+I255A+P256K；
(n)A150G+E210V+T231R+N233R+I255A+P256K；
(o)I202L+E210G+T231R+N233R+I255A+P256K；
(p)E1N+A18K+V60K+I86V+A150G+E210A+L227G+T231R+N233R+P256K；
(q)E1L+D27K+V60K+I86V+A150G+S219P+L227G+T231R+N233R+P256K；
(r)E1N+S58A+V60S+S83T+A150G+L227G+T231R+N233R+I255A+P256K；
(s)E1N+S58T+V60K+I86V+D102A+T143S+A150G+L227G+T231R+N233R+I255A+P256K；
(t)E1N+S58A+V60S+I86V+K98I+E99K+D102A+T143S+A150G+S216P+L227G+T231R+N233R+I255A+P256K；和
(u)S58A+V60S+S83T+A150A+L227G+T231R+N233R+I255A+P256K。
12.前述权利要求中任一所述的多肽，其中所述多肽与SEQ ID NO：2是至少50％，至少60％，至少70％，至少80％，至少85％，至少90％，至少95％，至少96％，至少97％，至少98％，至少99％或100％相同的。
13.一种编码权利要求1-12任一所述的多肽的分离的多核苷酸。
14.一种包含权利要求13的多核苷酸的核酸构建体，所述多核苷酸可操作连接于至少一个在表达宿主中指导所述多肽产生的调控序列。
15.一种包含权利要求14的核酸构建体的重组表达载体。
16.一种包含权利要求14的核酸构建体或权利要求15的重组表达载体的转化的宿主细胞。
17.一种制备权利要求1-12任一所述的多肽的方法，包括下述步骤：
(a)在有益于所述多肽产生的条件下培养转化的宿主细胞，所述宿主细胞包含含有所述多肽的重组表达载体或核酸构建体；和
(b)回收该多肽。
18.一种包含权利要求1-12任一所述的多肽的配制物。
19.权利要求18的配制物，其中所述配制物可为固体或液体配制物。
20.一种通过使用权利要求1-12任一所述的多肽在脂质水解过程中减少产生气味的短链脂肪酸形成的方法。
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US3244308P | 2008-02-29 | 2008-02-29 | |
| EP08152163 | 2008-02-29 | ||
| EP08152163.5 | 2008-02-29 | ||
| US61/032,443 | 2008-02-29 | ||
| PCT/EP2009/052246 WO2009109500A1 (en) | 2008-02-29 | 2009-02-26 | Polypeptides having lipase activity and polynucleotides encoding same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101960008A true CN101960008A (zh) | 2011-01-26 |
| CN101960008B CN101960008B (zh) | 2016-04-13 |
Family
ID=39745667
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200980106985.3A Active CN101960008B (zh) | 2008-02-29 | 2009-02-26 | 具有脂肪酶活性的多肽和编码该多肽的多核苷酸 |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US7919298B2 (zh) |
| EP (1) | EP2250259B1 (zh) |
| JP (1) | JP5650543B2 (zh) |
| CN (1) | CN101960008B (zh) |
| AR (1) | AR070490A1 (zh) |
| CA (1) | CA2715829C (zh) |
| ES (1) | ES2603979T3 (zh) |
| MX (1) | MX2010009072A (zh) |
| WO (1) | WO2009109500A1 (zh) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293744A (zh) * | 2014-08-19 | 2015-01-21 | 浙江工业大学 | 来源于嗜热踝节菌的脂肪酶突变体及应用 |
| CN106459939A (zh) * | 2014-05-27 | 2017-02-22 | 诺维信公司 | 脂肪酶变体以及编码它们的多核苷酸 |
| CN110923215A (zh) * | 2018-09-19 | 2020-03-27 | 江苏师范大学 | 一种生产米黑根毛霉脂肪酶mRML酶粉的方法 |
Families Citing this family (273)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9163203B2 (en) * | 2008-02-29 | 2015-10-20 | Novozymes A/S | Lipolytic enzyme variant with improved stability and polynucleotides encoding same |
| EP2343310A1 (en) | 2010-01-08 | 2011-07-13 | Novozymes A/S | Serine hydrolase formulation |
| CN106978403B (zh) | 2010-04-26 | 2021-12-14 | 诺维信公司 | 酶颗粒剂 |
| CN107475235B (zh) | 2011-06-20 | 2022-09-13 | 诺维信公司 | 颗粒组合物 |
| EP2723858B1 (en) | 2011-06-24 | 2017-04-12 | Novozymes A/S | Polypeptides having protease activity and polynucleotides encoding same |
| BR112013033816A2 (pt) | 2011-07-01 | 2017-02-14 | Novozymes A / S | composição de detergente líquido sem boro |
| IN2014CN00700A (zh) | 2011-07-01 | 2015-04-03 | Novozymes As | |
| US10711262B2 (en) | 2011-07-12 | 2020-07-14 | Novozymes A/S | Storage-stable enzyme granules |
| CN103748219A (zh) | 2011-08-15 | 2014-04-23 | 诺维信公司 | 具有纤维素酶活性的多肽以及编码它的多核苷酸 |
| EP2751266B1 (en) | 2011-09-22 | 2017-03-29 | Novozymes A/S | Polypeptides having protease activity and polynucleotides encoding same |
| ES2624531T3 (es) | 2011-11-25 | 2017-07-14 | Novozymes A/S | Polipéptidos que tienen actividad de lisozima y polinucleótidos que codifican los mismos |
| WO2013076269A1 (en) | 2011-11-25 | 2013-05-30 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| EP2607468A1 (en) | 2011-12-20 | 2013-06-26 | Henkel AG & Co. KGaA | Detergent compositions comprising subtilase variants |
| MX2014007446A (es) | 2011-12-20 | 2014-08-01 | Novozymes As | Variantes de subtilasa y polinucleotidos que las codifican. |
| CN111187676A (zh) | 2011-12-29 | 2020-05-22 | 诺维信公司 | 具有脂肪酶变体的洗涤剂组合物 |
| US9801398B2 (en) | 2012-01-26 | 2017-10-31 | Novozymes A/S | Use of polypeptides having protease activity in animal feed and detergents |
| MX350713B (es) | 2012-02-17 | 2017-09-14 | Novozymes As | Variantes de subtilisina y polinucleotidos que las codifican. |
| ES2582608T3 (es) | 2012-02-17 | 2016-09-14 | Henkel Ag & Co. Kgaa | Composiciones detergentes que comprenden variantes de subtilasa |
| US20150064773A1 (en) | 2012-03-07 | 2015-03-05 | Novozymes A/S | Detergent Composition and Substitution of Optical Brighteners in Detergent Composition |
| US10087401B2 (en) | 2012-03-16 | 2018-10-02 | Monosol, Llc | Water soluble compositions incorporating enzymes, and method of making same |
| CN104204183A (zh) | 2012-03-29 | 2014-12-10 | 诺维信公司 | 酶用于制备水溶性膜的用途 |
| WO2013149858A1 (en) | 2012-04-02 | 2013-10-10 | Novozymes A/S | Lipase variants and polynucleotides encoding same |
| US9394092B2 (en) | 2012-04-16 | 2016-07-19 | Monosol, Llc | Powdered pouch and method of making same |
| AR090971A1 (es) | 2012-05-07 | 2014-12-17 | Novozymes As | Polipeptidos que tienen actividad de degradacion de xantano y polinucleotidos que los codifican |
| JP2015525248A (ja) | 2012-05-16 | 2015-09-03 | ノボザイムス アクティーゼルスカブ | リパーゼを含む組成物およびその使用方法 |
| US20150184208A1 (en) | 2012-06-19 | 2015-07-02 | Novozymes A/S | Enzymatic reduction of hydroperoxides |
| MX364390B (es) | 2012-06-20 | 2019-04-25 | Novozymes As | Uso de polipeptidos que tienen actividad proteasa en alimentos para animales y detergentes. |
| WO2014012506A1 (en) | 2012-07-18 | 2014-01-23 | Novozymes A/S | Method of treating polyester textile |
| PL3553172T3 (pl) | 2012-08-16 | 2023-03-27 | Novozymes A/S | Sposób obróbki wyrobu włókienniczego z użyciem endoglukanazy |
| WO2014068083A1 (en) | 2012-11-01 | 2014-05-08 | Novozymes A/S | Method for removal of dna |
| US10323217B2 (en) | 2012-12-07 | 2019-06-18 | Novozymes A/S | Detergent composition comprising enzymes and washing method for preventing adhesion of bacteria |
| WO2014090940A1 (en) | 2012-12-14 | 2014-06-19 | Novozymes A/S | Removal of skin-derived body soils |
| CN104869841A (zh) | 2012-12-21 | 2015-08-26 | 诺维信公司 | 具有蛋白酶活性的多肽和编码它的多核苷酸 |
| US9902946B2 (en) | 2013-01-03 | 2018-02-27 | Novozymes A/S | Alpha-amylase variants and polynucleotides encoding same |
| US20160024440A1 (en) | 2013-03-14 | 2016-01-28 | Novozymes A/S | Enzyme and Inhibitor Containing Water-Soluble Films |
| CN105051174B (zh) | 2013-03-21 | 2018-04-03 | 诺维信公司 | 具有脂肪酶活性的多肽和编码它们的多核苷酸 |
| CN105164147B (zh) | 2013-04-23 | 2020-03-03 | 诺维信公司 | 具有稳定的枯草杆菌蛋白酶的液体自动餐具清洗洗涤剂组合物 |
| WO2014177709A1 (en) | 2013-05-03 | 2014-11-06 | Novozymes A/S | Microencapsulation of detergent enzymes |
| BR112015028666B8 (pt) | 2013-05-14 | 2022-08-09 | Novozymes As | Composição detergente, método para produzir a mesma, método para a limpeza de um objeto e usos da composição |
| US20160083703A1 (en) | 2013-05-17 | 2016-03-24 | Novozymes A/S | Polypeptides having alpha amylase activity |
| CN114634921A (zh) | 2013-06-06 | 2022-06-17 | 诺维信公司 | α-淀粉酶变体以及对其进行编码的多核苷酸 |
| WO2014207224A1 (en) | 2013-06-27 | 2014-12-31 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| WO2014207227A1 (en) | 2013-06-27 | 2014-12-31 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| CN105358670A (zh) | 2013-07-04 | 2016-02-24 | 诺维信公司 | 具有抗再沉积效果的具黄原胶裂解酶活性的多肽与编码它们的多核苷酸 |
| EP3019603A1 (en) | 2013-07-09 | 2016-05-18 | Novozymes A/S | Polypeptides with lipase activity and polynucleotides encoding same |
| RU2670946C9 (ru) | 2013-07-29 | 2018-11-26 | Новозимс А/С | Варианты протеазы и кодирующие их полинуклеотиды |
| WO2015014803A1 (en) | 2013-07-29 | 2015-02-05 | Novozymes A/S | Protease variants and polynucleotides encoding same |
| EP3339436B1 (en) | 2013-07-29 | 2021-03-31 | Henkel AG & Co. KGaA | Detergent composition comprising protease variants |
| WO2015049370A1 (en) | 2013-10-03 | 2015-04-09 | Novozymes A/S | Detergent composition and use of detergent composition |
| EP3074509B1 (en) | 2013-11-29 | 2019-03-06 | Novozymes A/S | Peroxygenase variants |
| EP3453757B1 (en) | 2013-12-20 | 2020-06-17 | Novozymes A/S | Polypeptides having protease activity and polynucleotides encoding same |
| WO2015109972A1 (en) | 2014-01-22 | 2015-07-30 | Novozymes A/S | Polypeptides with lipase activity and polynucleotides encoding same |
| EP3114272A1 (en) | 2014-03-05 | 2017-01-11 | Novozymes A/S | Compositions and methods for improving properties of cellulosic textile materials with xyloglucan endotransglycosylase |
| EP3114219A1 (en) | 2014-03-05 | 2017-01-11 | Novozymes A/S | Compositions and methods for improving properties of non-cellulosic textile materials with xyloglucan endotransglycosylase |
| CN106103721B (zh) | 2014-03-12 | 2020-01-03 | 诺维信公司 | 具有脂肪酶活性的多肽和编码它们的多核苷酸 |
| CN106103708A (zh) | 2014-04-01 | 2016-11-09 | 诺维信公司 | 具有α淀粉酶活性的多肽 |
| US10131863B2 (en) | 2014-04-11 | 2018-11-20 | Novozymes A/S | Detergent composition |
| WO2015158237A1 (en) | 2014-04-15 | 2015-10-22 | Novozymes A/S | Polypeptides with lipase activity and polynucleotides encoding same |
| EP3878957A1 (en) | 2014-05-27 | 2021-09-15 | Novozymes A/S | Methods for producing lipases |
| CN106414729A (zh) | 2014-06-12 | 2017-02-15 | 诺维信公司 | α‑淀粉酶变体以及对其进行编码的多核苷酸 |
| US20170121646A1 (en) | 2014-07-03 | 2017-05-04 | Novozymes A/S | Improved Stabilization of Non-Protease Enzyme |
| EP3878960A1 (en) | 2014-07-04 | 2021-09-15 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| WO2016001450A2 (en) | 2014-07-04 | 2016-01-07 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| WO2016079110A2 (en) | 2014-11-19 | 2016-05-26 | Novozymes A/S | Use of enzyme for cleaning |
| US10287562B2 (en) | 2014-11-20 | 2019-05-14 | Novoszymes A/S | Alicyclobacillus variants and polynucleotides encoding same |
| WO2016087617A1 (en) | 2014-12-04 | 2016-06-09 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| WO2016087401A1 (en) | 2014-12-05 | 2016-06-09 | Novozymes A/S | Lipase variants and polynucleotides encoding same |
| WO2016096714A1 (en) | 2014-12-15 | 2016-06-23 | Henkel Ag & Co. Kgaa | Detergent composition comprising subtilase variants |
| US20180000076A1 (en) | 2014-12-16 | 2018-01-04 | Novozymes A/S | Polypeptides Having N-Acetyl Glucosamine Oxidase Activity |
| WO2016097350A1 (en) | 2014-12-19 | 2016-06-23 | Novozymes A/S | Protease variants and polynucleotides encoding same |
| DK3234093T3 (da) | 2014-12-19 | 2020-08-17 | Novozymes As | Proteasevarianter og polynukleotider, som koder for dem |
| CN107864658A (zh) | 2015-04-07 | 2018-03-30 | 诺维信公司 | 用于选择具有脂肪酶活性的酶的方法 |
| US20180105772A1 (en) | 2015-04-10 | 2018-04-19 | Novozymes A/S | Detergent composition |
| WO2016162556A1 (en) | 2015-04-10 | 2016-10-13 | Novozymes A/S | Laundry method, use of dnase and detergent composition |
| WO2016184944A1 (en) | 2015-05-19 | 2016-11-24 | Novozymes A/S | Odor reduction |
| ES2665989T3 (es) | 2015-06-04 | 2018-04-30 | The Procter & Gamble Company | Composición detergente líquida para lavado de vajillas a mano |
| ES2712459T3 (es) | 2015-06-04 | 2019-05-13 | Procter & Gamble | Composición detergente líquida para lavado de vajillas a mano |
| EP3310908B1 (en) | 2015-06-16 | 2020-08-05 | Novozymes A/S | Polypeptides with lipase activity and polynucleotides encoding same |
| WO2016202785A1 (en) | 2015-06-17 | 2016-12-22 | Novozymes A/S | Container |
| EP3106508B1 (en) | 2015-06-18 | 2019-11-20 | Henkel AG & Co. KGaA | Detergent composition comprising subtilase variants |
| CN108012544A (zh) | 2015-06-18 | 2018-05-08 | 诺维信公司 | 枯草杆菌酶变体以及编码它们的多核苷酸 |
| EP3317388B1 (en) | 2015-06-30 | 2019-11-13 | Novozymes A/S | Laundry detergent composition, method for washing and use of composition |
| EP3929285A3 (en) | 2015-07-01 | 2022-05-25 | Novozymes A/S | Methods of reducing odor |
| EP3320089B1 (en) | 2015-07-06 | 2021-06-16 | Novozymes A/S | Lipase variants and polynucleotides encoding same |
| KR20180053365A (ko) | 2015-09-17 | 2018-05-21 | 헨켈 아게 운트 코. 카게아아 | 크산탄 분해 활성을 갖는 폴리펩티드를 포함하는 세제 조성물 |
| CA2991114A1 (en) | 2015-09-17 | 2017-03-23 | Novozymes A/S | Polypeptides having xanthan degrading activity and polynucleotides encoding same |
| US10934535B2 (en) | 2015-10-07 | 2021-03-02 | Novozymes A/S | Detergent composition comprising a DNase |
| CN108291215A (zh) | 2015-10-14 | 2018-07-17 | 诺维信公司 | 具有蛋白酶活性的多肽以及编码它们的多核苷酸 |
| EP3362168A1 (en) | 2015-10-14 | 2018-08-22 | Novozymes A/S | Cleaning of water filtration membranes |
| CN108291212A (zh) | 2015-10-14 | 2018-07-17 | 诺维信公司 | 多肽变体 |
| WO2016203064A2 (en) | 2015-10-28 | 2016-12-22 | Novozymes A/S | Detergent composition comprising protease and amylase variants |
| US11001821B2 (en) | 2015-11-24 | 2021-05-11 | Novozymes A/S | Polypeptides having protease activity and polynucleotides encoding same |
| CN108431217B (zh) | 2015-12-01 | 2022-06-21 | 诺维信公司 | 用于产生脂肪酶的方法 |
| EP3397061A1 (en) | 2015-12-28 | 2018-11-07 | Novozymes BioAG A/S | Heat priming of bacterial spores |
| EP3715442A1 (en) | 2016-03-23 | 2020-09-30 | Novozymes A/S | Use of polypeptide having dnase activity for treating fabrics |
| US20200325418A1 (en) | 2016-04-08 | 2020-10-15 | Novozymes A/S | Detergent compositions and uses of the same |
| EP3693449A1 (en) | 2016-04-29 | 2020-08-12 | Novozymes A/S | Detergent compositions and uses thereof |
| WO2017210188A1 (en) | 2016-05-31 | 2017-12-07 | Novozymes A/S | Stabilized liquid peroxide compositions |
| CN109715792A (zh) | 2016-06-03 | 2019-05-03 | 诺维信公司 | 枯草杆菌酶变体和对其进行编码的多核苷酸 |
| EP3475404A1 (en) | 2016-06-23 | 2019-05-01 | Novozymes A/S | Use of enzymes, composition and method for removing soil |
| BR112018077510A2 (pt) | 2016-06-30 | 2019-04-02 | Novozymes As | variantes de lipase e composições que compreendem tensoativo e variante de lipase |
| WO2018002261A1 (en) | 2016-07-01 | 2018-01-04 | Novozymes A/S | Detergent compositions |
| WO2018007573A1 (en) | 2016-07-08 | 2018-01-11 | Novozymes A/S | Detergent compositions with galactanase |
| JP6858850B2 (ja) | 2016-07-13 | 2021-04-14 | ザ プロクター アンド ギャンブル カンパニーＴｈｅ Ｐｒｏｃｔｅｒ ＆ Ｇａｍｂｌｅ Ｃｏｍｐａｎｙ | バチルス・シビ（ＢＡＣＩＬＬＵＳ ＣＩＢＩ）ＤＮａｓｅ変異体及びその使用 |
| US11326152B2 (en) | 2016-07-18 | 2022-05-10 | Novozymes A/S | Lipase variants, polynucleotides encoding same and the use thereof |
| ES2790148T3 (es) | 2016-08-17 | 2020-10-27 | Procter & Gamble | Composición limpiadora que comprende enzimas |
| AU2017317563B8 (en) | 2016-08-24 | 2023-03-23 | Henkel Ag & Co. Kgaa | Detergent compositions comprising xanthan lyase variants I |
| US11072765B2 (en) | 2016-08-24 | 2021-07-27 | Novozymes A/S | GH9 endoglucanase variants and polynucleotides encoding same |
| CA3032248A1 (en) | 2016-08-24 | 2018-03-01 | Novozymes A/S | Xanthan lyase variants and polynucleotides encoding same |
| AU2017317564B2 (en) | 2016-08-24 | 2021-09-30 | Henkel Ag & Co. Kgaa | Detergent composition comprising GH9 endoglucanase variants I |
| CN110023474A (zh) | 2016-09-29 | 2019-07-16 | 诺维信公司 | 酶用于洗涤的用途、洗涤方法和器皿洗涤组合物 |
| WO2018060475A1 (en) | 2016-09-29 | 2018-04-05 | Novozymes A/S | Spore containing granule |
| US20210284933A1 (en) | 2016-10-25 | 2021-09-16 | Novozymes A/S | Detergent compositions |
| CN110072986B (zh) | 2016-11-01 | 2023-04-04 | 诺维信公司 | 多核心颗粒 |
| CA3043443A1 (en) | 2016-12-01 | 2018-06-07 | Basf Se | Stabilization of enzymes in compositions |
| EP3551740B1 (en) | 2016-12-12 | 2021-08-11 | Novozymes A/S | Use of polypeptides |
| WO2018177936A1 (en) | 2017-03-31 | 2018-10-04 | Novozymes A/S | Polypeptides having dnase activity |
| CN110651039A (zh) | 2017-03-31 | 2020-01-03 | 诺维信公司 | 具有rna酶活性的多肽 |
| EP3601550A1 (en) | 2017-03-31 | 2020-02-05 | Novozymes A/S | Polypeptides having dnase activity |
| EP3607039A1 (en) | 2017-04-04 | 2020-02-12 | Novozymes A/S | Polypeptides |
| US11339355B2 (en) | 2017-04-04 | 2022-05-24 | Novozymes A/S | Glycosyl hydrolases |
| EP3607040A1 (en) | 2017-04-04 | 2020-02-12 | Novozymes A/S | Polypeptide compositions and uses thereof |
| EP3385361B1 (en) | 2017-04-05 | 2019-03-27 | Henkel AG & Co. KGaA | Detergent compositions comprising bacterial mannanases |
| EP3385362A1 (en) | 2017-04-05 | 2018-10-10 | Henkel AG & Co. KGaA | Detergent compositions comprising fungal mannanases |
| WO2018184818A1 (en) | 2017-04-06 | 2018-10-11 | Novozymes A/S | Cleaning compositions and uses thereof |
| US11407964B2 (en) | 2017-04-06 | 2022-08-09 | Novozymes A/S | Cleaning compositions and uses thereof |
| MX2019011764A (es) | 2017-04-06 | 2019-11-28 | Novozymes As | Composiciones limpiadoras y usos de las mismas. |
| EP3607060B1 (en) | 2017-04-06 | 2021-08-11 | Novozymes A/S | Detergent compositions and uses thereof |
| US20200190438A1 (en) | 2017-04-06 | 2020-06-18 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2018184816A1 (en) | 2017-04-06 | 2018-10-11 | Novozymes A/S | Cleaning compositions and uses thereof |
| CN110662829B (zh) | 2017-04-06 | 2022-03-01 | 诺维信公司 | 清洁组合物及其用途 |
| EP3607042A1 (en) | 2017-04-06 | 2020-02-12 | Novozymes A/S | Cleaning compositions and uses thereof |
| CN110651038A (zh) | 2017-05-05 | 2020-01-03 | 诺维信公司 | 包含脂肪酶和亚硫酸盐的组合物 |
| EP3401385A1 (en) | 2017-05-08 | 2018-11-14 | Henkel AG & Co. KGaA | Detergent composition comprising polypeptide comprising carbohydrate-binding domain |
| WO2018206535A1 (en) | 2017-05-08 | 2018-11-15 | Novozymes A/S | Carbohydrate-binding domain and polynucleotides encoding the same |
| CN111108183A (zh) | 2017-06-30 | 2020-05-05 | 诺维信公司 | 酶浆液组合物 |
| WO2019038060A1 (en) | 2017-08-24 | 2019-02-28 | Henkel Ag & Co. Kgaa | DETERGENT COMPOSITION COMPRISING XANTHANE LYASE II VARIANTS |
| US11624059B2 (en) | 2017-08-24 | 2023-04-11 | Henkel Ag & Co. Kgaa | Detergent compositions comprising GH9 endoglucanase variants II |
| CN111278971A (zh) | 2017-08-24 | 2020-06-12 | 诺维信公司 | Gh9内切葡聚糖酶变体以及编码它们的多核苷酸 |
| US11359188B2 (en) | 2017-08-24 | 2022-06-14 | Novozymes A/S | Xanthan lyase variants and polynucleotides encoding same |
| MX2020002953A (es) | 2017-09-20 | 2020-07-22 | Novozymes As | Uso de enzimas para mejorar la absorcion de agua y/o la blancura. |
| US11414814B2 (en) | 2017-09-22 | 2022-08-16 | Novozymes A/S | Polypeptides |
| JP7114697B2 (ja) | 2017-09-27 | 2022-08-08 | ザ プロクター アンド ギャンブル カンパニー | リパーゼを含む洗剤組成物 |
| MX2020002964A (es) * | 2017-09-27 | 2020-07-22 | Novozymes As | Variantes de lipasa y composiciones de microcapsula que comprenden las variantes de lipasa. |
| WO2019076800A1 (en) | 2017-10-16 | 2019-04-25 | Novozymes A/S | CLEANING COMPOSITIONS AND USES THEREOF |
| CN111448302A (zh) | 2017-10-16 | 2020-07-24 | 诺维信公司 | 低粉化颗粒 |
| WO2019076834A1 (en) | 2017-10-16 | 2019-04-25 | Novozymes A/S | PELLETS WITH LOW DUST CONTENT |
| WO2019081724A1 (en) | 2017-10-27 | 2019-05-02 | Novozymes A/S | VARIANTS OF DNASE |
| PL3476935T3 (pl) | 2017-10-27 | 2022-03-28 | The Procter & Gamble Company | Kompozycje detergentowe zawierające odmiany polipeptydowe |
| US11505767B2 (en) | 2017-11-01 | 2022-11-22 | Novozymes A/S | Methods for cleansing medical devices |
| WO2019086528A1 (en) | 2017-11-01 | 2019-05-09 | Novozymes A/S | Polypeptides and compositions comprising such polypeptides |
| US11767492B2 (en) | 2017-11-01 | 2023-09-26 | Novozymes A/S | Methods of treating fabric using a Lactobacillus hexosaminidase |
| DE102017125560A1 (de) | 2017-11-01 | 2019-05-02 | Henkel Ag & Co. Kgaa | Reinigungszusammensetzungen, die dispersine iii enthalten |
| DE102017125558A1 (de) | 2017-11-01 | 2019-05-02 | Henkel Ag & Co. Kgaa | Reinigungszusammensetzungen, die dispersine i enthalten |
| DE102017125559A1 (de) | 2017-11-01 | 2019-05-02 | Henkel Ag & Co. Kgaa | Reinigungszusammensetzungen, die dispersine ii enthalten |
| CA3083390A1 (en) | 2017-11-29 | 2019-06-06 | Basf Se | Storage-stable enzyme preparations, their production and use |
| WO2019110462A1 (en) | 2017-12-04 | 2019-06-13 | Novozymes A/S | Lipase variants and polynucleotides encoding same |
| JP7309718B2 (ja) | 2017-12-20 | 2023-07-18 | ビーエーエスエフ ソシエタス・ヨーロピア | 繊維製品に付着した、融解温度が３０℃超のである脂肪化合物を除去するための洗濯用配合物 |
| US20210123033A1 (en) | 2018-02-08 | 2021-04-29 | Novozymes A/S | Lipases, Lipase Variants and Compositions Thereof |
| US20210071155A1 (en) | 2018-02-08 | 2021-03-11 | Novozymes A/S | Lipase Variants and Compositions Thereof |
| EP3755793A1 (en) | 2018-02-23 | 2020-12-30 | Henkel AG & Co. KGaA | Detergent composition comprising xanthan lyase and endoglucanase variants |
| WO2019175240A1 (en) | 2018-03-13 | 2019-09-19 | Novozymes A/S | Microencapsulation using amino sugar oligomers |
| US20210009979A1 (en) | 2018-03-23 | 2021-01-14 | Novozymes A/S | Subtilase variants and compositions comprising same |
| WO2019201793A1 (en) | 2018-04-17 | 2019-10-24 | Novozymes A/S | Polypeptides comprising carbohydrate binding activity in detergent compositions and their use in reducing wrinkles in textile or fabric. |
| EP3781680A1 (en) | 2018-04-19 | 2021-02-24 | Novozymes A/S | Stabilized cellulase variants |
| CN111989389A (zh) | 2018-04-19 | 2020-11-24 | 巴斯夫欧洲公司 | 组合物以及可用于该组合物的聚合物 |
| US11661592B2 (en) | 2018-04-19 | 2023-05-30 | Novozymes A/S | Stabilized endoglucanase variants |
| WO2019238761A1 (en) | 2018-06-15 | 2019-12-19 | Basf Se | Water soluble multilayer films containing wash active chemicals and enzymes |
| WO2020002604A1 (en) | 2018-06-28 | 2020-01-02 | Novozymes A/S | Detergent compositions and uses thereof |
| US20210189297A1 (en) | 2018-06-29 | 2021-06-24 | Novozymes A/S | Subtilase variants and compositions comprising same |
| EP3814473A1 (en) | 2018-06-29 | 2021-05-05 | Novozymes A/S | Detergent compositions and uses thereof |
| US20210189296A1 (en) | 2018-07-02 | 2021-06-24 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020007875A1 (en) | 2018-07-03 | 2020-01-09 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020008043A1 (en) | 2018-07-06 | 2020-01-09 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020008024A1 (en) | 2018-07-06 | 2020-01-09 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020030623A1 (en) | 2018-08-10 | 2020-02-13 | Basf Se | Packaging unit comprising a detergent composition containing an enzyme and at least one chelating agent |
| US20210340466A1 (en) | 2018-10-01 | 2021-11-04 | Novozymes A/S | Detergent compositions and uses thereof |
| WO2020070209A1 (en) | 2018-10-02 | 2020-04-09 | Novozymes A/S | Cleaning composition |
| WO2020070014A1 (en) | 2018-10-02 | 2020-04-09 | Novozymes A/S | Cleaning composition comprising anionic surfactant and a polypeptide having rnase activity |
| US20230287306A1 (en) | 2018-10-02 | 2023-09-14 | Novozymes A/S | Cleaning Composition |
| WO2020070199A1 (en) | 2018-10-03 | 2020-04-09 | Novozymes A/S | Polypeptides having alpha-mannan degrading activity and polynucleotides encoding same |
| WO2020070249A1 (en) | 2018-10-03 | 2020-04-09 | Novozymes A/S | Cleaning compositions |
| WO2020069915A1 (en) | 2018-10-05 | 2020-04-09 | Basf Se | Compounds stabilizing hydrolases in liquids |
| MX2021003931A (es) | 2018-10-05 | 2021-06-04 | Basf Se | Compuestos estabilizadores de amilasas en liquidos. |
| EP3677676A1 (en) | 2019-01-03 | 2020-07-08 | Basf Se | Compounds stabilizing amylases in liquids |
| BR112021005954A2 (pt) | 2018-10-05 | 2021-06-29 | Basf Se | preparação enzimática, processo para produzir uma preparação enzimática estável, métodos para reduzir perda de atividade lipolítica de pelo menos uma lipase, para preparar uma formulação de detergente, para remover manchas e para aumentar a estabilidade de armazenagem de uma formulação de detergente líquido, usos de um composto e da preparação enzimática, e, formulação de detergente. |
| EP3864124A1 (en) | 2018-10-11 | 2021-08-18 | Novozymes A/S | Cleaning compositions and uses thereof |
| EP3647398B1 (en) | 2018-10-31 | 2024-05-15 | Henkel AG & Co. KGaA | Cleaning compositions containing dispersins v |
| EP3647397A1 (en) | 2018-10-31 | 2020-05-06 | Henkel AG & Co. KGaA | Cleaning compositions containing dispersins iv |
| WO2020104231A1 (en) | 2018-11-19 | 2020-05-28 | Basf Se | Powders and granules containing a chelating agent and an enzyme |
| WO2020114965A1 (en) | 2018-12-03 | 2020-06-11 | Novozymes A/S | LOW pH POWDER DETERGENT COMPOSITION |
| EP3891277A1 (en) | 2018-12-03 | 2021-10-13 | Novozymes A/S | Powder detergent compositions |
| WO2020127796A2 (en) | 2018-12-21 | 2020-06-25 | Novozymes A/S | Polypeptides having peptidoglycan degrading activity and polynucleotides encoding same |
| EP3898919A1 (en) | 2018-12-21 | 2021-10-27 | Novozymes A/S | Detergent pouch comprising metalloproteases |
| EP3702452A1 (en) | 2019-03-01 | 2020-09-02 | Novozymes A/S | Detergent compositions comprising two proteases |
| US20220177808A1 (en) | 2019-03-08 | 2022-06-09 | Basf Se | Cationic surfactant and its use in laundry detergent compositions |
| EP3942032A1 (en) | 2019-03-21 | 2022-01-26 | Novozymes A/S | Alpha-amylase variants and polynucleotides encoding same |
| CN113785039A (zh) | 2019-04-03 | 2021-12-10 | 诺维信公司 | 具有β-葡聚糖酶活性的多肽、编码其的多核苷酸及其在清洁和洗涤剂组合物中的用途 |
| EP3953462A1 (en) | 2019-04-10 | 2022-02-16 | Novozymes A/S | Polypeptide variants |
| US20220186151A1 (en) | 2019-04-12 | 2022-06-16 | Novozymes A/S | Stabilized glycoside hydrolase variants |
| WO2020229480A1 (en) | 2019-05-14 | 2020-11-19 | Basf Se | Compounds stabilizing hydrolases in liquids |
| CN110904074B (zh) * | 2019-05-31 | 2020-12-01 | 江南大学 | 一种脂肪酶突变体及其在去污方面的应用 |
| JP2022538360A (ja) | 2019-07-01 | 2022-09-01 | ビーエーエスエフ ソシエタス・ヨーロピア | 酵素を安定化するためのペプチドアセタール |
| EP3994255A1 (en) | 2019-07-02 | 2022-05-11 | Novozymes A/S | Lipase variants and compositions thereof |
| US20220403298A1 (en) | 2019-07-12 | 2022-12-22 | Novozymes A/S | Enzymatic emulsions for detergents |
| WO2021037895A1 (en) | 2019-08-27 | 2021-03-04 | Novozymes A/S | Detergent composition |
| EP4022020A1 (en) | 2019-08-27 | 2022-07-06 | Novozymes A/S | Composition comprising a lipase |
| WO2021053127A1 (en) | 2019-09-19 | 2021-03-25 | Novozymes A/S | Detergent composition |
| WO2021064068A1 (en) | 2019-10-03 | 2021-04-08 | Novozymes A/S | Polypeptides comprising at least two carbohydrate binding domains |
| CN114585718A (zh) | 2019-10-18 | 2022-06-03 | 巴斯夫欧洲公司 | 储存稳定的包含水解酶的液体 |
| WO2021105330A1 (en) | 2019-11-29 | 2021-06-03 | Basf Se | Compositions and polymers useful for such compositions |
| WO2021115912A1 (en) | 2019-12-09 | 2021-06-17 | Basf Se | Formulations comprising a hydrophobically modified polyethyleneimine and one or more enzymes |
| AU2020404593A1 (en) | 2019-12-20 | 2022-08-18 | Henkel Ag & Co. Kgaa | Cleaning compositions comprising dispersins VI |
| WO2021123307A2 (en) | 2019-12-20 | 2021-06-24 | Novozymes A/S | Polypeptides having proteolytic activity and use thereof |
| US20220411726A1 (en) | 2019-12-20 | 2022-12-29 | Novozymes A/S | Stabilized liquid boron-free enzyme compositions |
| EP4077619A1 (en) | 2019-12-20 | 2022-10-26 | Henkel AG & Co. KGaA | Cleaning composition coprising a dispersin and a carbohydrase |
| AU2020405786A1 (en) | 2019-12-20 | 2022-08-11 | Henkel Ag & Co. Kgaa | Cleaning compositions comprising dispersins IX |
| EP4077621A2 (en) | 2019-12-20 | 2022-10-26 | Henkel AG & Co. KGaA | Cleaning compositions comprising dispersins viii |
| EP3892708A1 (en) | 2020-04-06 | 2021-10-13 | Henkel AG & Co. KGaA | Cleaning compositions comprising dispersin variants |
| US20230143128A1 (en) | 2020-04-08 | 2023-05-11 | Novozymes A/S | Carbohydrate binding module variants |
| CN115516071A (zh) | 2020-04-21 | 2022-12-23 | 诺维信公司 | 包含具有果聚糖降解活性的多肽的清洁组合物 |
| EP3907271A1 (en) | 2020-05-07 | 2021-11-10 | Novozymes A/S | Cleaning composition, use and method of cleaning |
| US20230212548A1 (en) | 2020-05-26 | 2023-07-06 | Novozymes A/S | Subtilase variants and compositions comprising same |
| EP4168523A1 (en) | 2020-06-18 | 2023-04-26 | Basf Se | Compositions and their use |
| EP4172298A1 (en) | 2020-06-24 | 2023-05-03 | Novozymes A/S | Use of cellulases for removing dust mite from textile |
| EP3936593A1 (en) | 2020-07-08 | 2022-01-12 | Henkel AG & Co. KGaA | Cleaning compositions and uses thereof |
| US20230137116A1 (en) | 2020-07-09 | 2023-05-04 | Basf Se | Compositions and their applications |
| WO2022008732A1 (en) | 2020-07-10 | 2022-01-13 | Basf Se | Enhancing the activity of antimicrobial preservatives |
| CN116057158A (zh) | 2020-07-27 | 2023-05-02 | 联合利华知识产权控股有限公司 | 酶和表面活性剂用于抑制微生物的用途 |
| CN116323889A (zh) | 2020-08-25 | 2023-06-23 | 诺维信公司 | 家族44木葡聚糖酶变体 |
| EP4204547A1 (en) | 2020-08-28 | 2023-07-05 | Novozymes A/S | Protease variants with improved solubility |
| JP2023544111A (ja) | 2020-09-22 | 2023-10-20 | ビーエーエスエフ ソシエタス・ヨーロピア | 第二の酵素を含む、プロテアーゼとプロテアーゼ阻害薬との改善された組合せ |
| CN116507725A (zh) | 2020-10-07 | 2023-07-28 | 诺维信公司 | α-淀粉酶变体 |
| WO2022083949A1 (en) | 2020-10-20 | 2022-04-28 | Basf Se | Compositions and their use |
| WO2022084303A2 (en) | 2020-10-20 | 2022-04-28 | Novozymes A/S | Use of polypeptides having dnase activity |
| CN116615523A (zh) | 2020-10-28 | 2023-08-18 | 诺维信公司 | 脂氧合酶的用途 |
| BR112023008326A2 (pt) | 2020-10-29 | 2023-12-12 | Novozymes As | Variantes de lipase e composições compreendendo tais variantes de lipase |
| WO2022103725A1 (en) | 2020-11-13 | 2022-05-19 | Novozymes A/S | Detergent composition comprising a lipase |
| WO2022106400A1 (en) | 2020-11-18 | 2022-05-27 | Novozymes A/S | Combination of immunochemically different proteases |
| WO2022106404A1 (en) | 2020-11-18 | 2022-05-27 | Novozymes A/S | Combination of proteases |
| EP4032966A1 (en) | 2021-01-22 | 2022-07-27 | Novozymes A/S | Liquid enzyme composition with sulfite scavenger |
| EP4284905A1 (en) | 2021-01-28 | 2023-12-06 | Novozymes A/S | Lipase with low malodor generation |
| EP4039806A1 (en) | 2021-02-04 | 2022-08-10 | Henkel AG & Co. KGaA | Detergent composition comprising xanthan lyase and endoglucanase variants with im-proved stability |
| WO2022171872A1 (en) | 2021-02-12 | 2022-08-18 | Novozymes A/S | Stabilized biological detergents |
| WO2022171780A2 (en) | 2021-02-12 | 2022-08-18 | Novozymes A/S | Alpha-amylase variants |
| EP4047088A1 (en) | 2021-02-22 | 2022-08-24 | Basf Se | Amylase variants |
| WO2022175435A1 (en) | 2021-02-22 | 2022-08-25 | Basf Se | Amylase variants |
| WO2022189521A1 (en) | 2021-03-12 | 2022-09-15 | Novozymes A/S | Polypeptide variants |
| EP4060036A1 (en) | 2021-03-15 | 2022-09-21 | Novozymes A/S | Polypeptide variants |
| WO2022194673A1 (en) | 2021-03-15 | 2022-09-22 | Novozymes A/S | Dnase variants |
| WO2022199418A1 (en) | 2021-03-26 | 2022-09-29 | Novozymes A/S | Detergent composition with reduced polymer content |
| EP4359518A1 (en) | 2021-06-23 | 2024-05-01 | Novozymes A/S | Alpha-amylase polypeptides |
| WO2023061827A1 (en) | 2021-10-13 | 2023-04-20 | Basf Se | Compositions comprising polymers, polymers, and their use |
| WO2023088777A1 (en) | 2021-11-22 | 2023-05-25 | Basf Se | Compositions comprising polymers, polymers, and their use |
| WO2023117989A1 (en) | 2021-12-21 | 2023-06-29 | Basf Se | Environmental attributes for formulation additives |
| WO2023116569A1 (en) | 2021-12-21 | 2023-06-29 | Novozymes A/S | Composition comprising a lipase and a booster |
| EP4206309A1 (en) | 2021-12-30 | 2023-07-05 | Novozymes A/S | Protein particles with improved whiteness |
| WO2023148086A1 (en) | 2022-02-04 | 2023-08-10 | Basf Se | Compositions comprising polymers, polymers, and their use |
| EP4234664A1 (en) | 2022-02-24 | 2023-08-30 | Evonik Operations GmbH | Composition comprising glucolipids and enzymes |
| WO2023165507A1 (en) | 2022-03-02 | 2023-09-07 | Novozymes A/S | Use of xyloglucanase for improvement of sustainability of detergents |
| WO2023165950A1 (en) | 2022-03-04 | 2023-09-07 | Novozymes A/S | Dnase variants and compositions |
| WO2023194204A1 (en) | 2022-04-08 | 2023-10-12 | Novozymes A/S | Hexosaminidase variants and compositions |
| WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
| DE102022205591A1 (de) | 2022-06-01 | 2023-12-07 | Henkel Ag & Co. Kgaa | Wasch- und reinigungsmittel mit verbesserter enzymstabilität |
| DE102022205593A1 (de) | 2022-06-01 | 2023-12-07 | Henkel Ag & Co. Kgaa | Wasch- und reinigungsmittel mit verbesserter enzymstabilität |
| DE102022205588A1 (de) | 2022-06-01 | 2023-12-07 | Henkel Ag & Co. Kgaa | Wasch- und reinigungsmittel mit verbesserter enzymstabilität |
| WO2023247664A2 (en) | 2022-06-24 | 2023-12-28 | Novozymes A/S | Lipase variants and compositions comprising such lipase variants |
| WO2024033136A1 (en) | 2022-08-11 | 2024-02-15 | Basf Se | Amylase variants |
| WO2024033135A2 (en) | 2022-08-11 | 2024-02-15 | Basf Se | Amylase variants |
| WO2024033134A1 (en) | 2022-08-11 | 2024-02-15 | Basf Se | Enzyme compositions comprising protease, mannanase, and/or cellulase |
| WO2024033133A2 (en) | 2022-08-11 | 2024-02-15 | Basf Se | Enzyme compositions comprising an amylase |
| EP4324900A1 (en) | 2022-08-17 | 2024-02-21 | Henkel AG & Co. KGaA | Detergent composition comprising enzymes |
| WO2024083589A1 (en) | 2022-10-18 | 2024-04-25 | Basf Se | Detergent compositions, polymers and methods of manufacturing the same |
Family Cites Families (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4810414A (en) | 1986-08-29 | 1989-03-07 | Novo Industri A/S | Enzymatic detergent additive |
| EP0305216B1 (en) | 1987-08-28 | 1995-08-02 | Novo Nordisk A/S | Recombinant Humicola lipase and process for the production of recombinant humicola lipases |
| GB8921995D0 (en) | 1989-09-29 | 1989-11-15 | Unilever Plc | Perfumed laundry detergents |
| JP4307549B2 (ja) | 1995-07-14 | 2009-08-05 | ノボザイムス アクティーゼルスカブ | 脂肪分解活性を有する修飾された酵素 |
| JP4068142B2 (ja) | 1995-08-11 | 2008-03-26 | ノボザイムス アクティーゼルスカブ | 新規の脂肪分解酵素 |
| US7312062B2 (en) * | 1998-11-27 | 2007-12-25 | Novozymes A/S | Lipolytic enzyme variants |
| NZ511340A (en) | 1998-11-27 | 2003-07-25 | Novozymes As | Lipolytic enzyme variants |
| WO2000060063A1 (en) | 1999-03-31 | 2000-10-12 | Novozymes A/S | Lipase variant |
| AU2002223020B2 (en) | 2000-11-27 | 2006-07-06 | Novozymes A/S | Automated mechanical stress assay for screening cleaning ingredients |
| US20040152180A1 (en) * | 2001-01-10 | 2004-08-05 | Steffan Minning | Lipolytic enzyme variant |
| DK1352057T3 (da) | 2001-01-10 | 2009-07-20 | Novozymes As | Upolytisk enzymvariant |
| AU2002229513A1 (en) | 2001-02-07 | 2002-08-19 | Novozymes A/S | Lipase variants |
| PL375355A1 (en) * | 2002-08-19 | 2005-11-28 | Dsm Ip Assets B.V. | Novel lipases and uses thereof |
| WO2006125437A2 (en) | 2005-05-27 | 2006-11-30 | Novozymes A/S | Swatch for testing the washing performance of an enzyme |
| EP2371949B1 (en) | 2006-01-23 | 2017-04-19 | Novozymes A/S | Lipase variants |
| WO2007087503A1 (en) * | 2006-01-23 | 2007-08-02 | Novozymes A/S | Polypeptides having lipase activity and polynucleotides encoding same |
-
2009
- 2009-02-26 CA CA2715829A patent/CA2715829C/en not_active Expired - Fee Related
- 2009-02-26 US US12/393,283 patent/US7919298B2/en active Active
- 2009-02-26 EP EP09716837.1A patent/EP2250259B1/en active Active
- 2009-02-26 ES ES09716837.1T patent/ES2603979T3/es active Active
- 2009-02-26 MX MX2010009072A patent/MX2010009072A/es active IP Right Grant
- 2009-02-26 CN CN200980106985.3A patent/CN101960008B/zh active Active
- 2009-02-26 AR ARP090100662A patent/AR070490A1/es unknown
- 2009-02-26 JP JP2010548100A patent/JP5650543B2/ja not_active Expired - Fee Related
- 2009-02-26 WO PCT/EP2009/052246 patent/WO2009109500A1/en active Application Filing
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106459939A (zh) * | 2014-05-27 | 2017-02-22 | 诺维信公司 | 脂肪酶变体以及编码它们的多核苷酸 |
| CN104293744A (zh) * | 2014-08-19 | 2015-01-21 | 浙江工业大学 | 来源于嗜热踝节菌的脂肪酶突变体及应用 |
| CN110923215A (zh) * | 2018-09-19 | 2020-03-27 | 江苏师范大学 | 一种生产米黑根毛霉脂肪酶mRML酶粉的方法 |
| CN110923215B (zh) * | 2018-09-19 | 2021-08-10 | 江苏师范大学 | 一种生产米黑根毛霉脂肪酶mRML酶粉的方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101960008B (zh) | 2016-04-13 |
| WO2009109500A8 (en) | 2010-10-14 |
| ES2603979T3 (es) | 2017-03-02 |
| AR070490A1 (es) | 2010-04-07 |
| WO2009109500A1 (en) | 2009-09-11 |
| JP2011512808A (ja) | 2011-04-28 |
| MX2010009072A (es) | 2010-09-24 |
| US7919298B2 (en) | 2011-04-05 |
| CA2715829A1 (en) | 2009-09-11 |
| EP2250259A1 (en) | 2010-11-17 |
| JP5650543B2 (ja) | 2015-01-07 |
| CA2715829C (en) | 2017-05-23 |
| EP2250259B1 (en) | 2016-08-31 |
| US20090221033A1 (en) | 2009-09-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101960008A (zh) | 具有脂肪酶活性的多肽和编码该多肽的多核苷酸 | |
| CN101370933B (zh) | 脂肪酶变体 | |
| CN101990573B (zh) | 具有改进的稳定性的脂肪分解酶变体及编码该变体的多核苷酸 | |
| CN101374947B (zh) | 具有脂肪酶活性的多肽和编码该多肽的多核苷酸 | |
| MX2008009207A (en) | Lipase variants | |
| BRPI0707202B1 (pt) | Variant, dna sequence, expression vector, trangenic microorganism, and, method of producing a variant of lipase | |
| MX2008008925A (es) | Polipeptidos que tienen actividad de lipasa y polinucleotidos que codifican para los mismos |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |