CN1306577A - 多点置换的蛋白酶变体 - Google Patents
多点置换的蛋白酶变体 Download PDFInfo
- Publication number
- CN1306577A CN1306577A CN98811436A CN98811436A CN1306577A CN 1306577 A CN1306577 A CN 1306577A CN 98811436 A CN98811436 A CN 98811436A CN 98811436 A CN98811436 A CN 98811436A CN 1306577 A CN1306577 A CN 1306577A
- Authority
- CN
- China
- Prior art keywords
- ease variants
- displacement
- subtilisin
- amino
- acid residue
- 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
- 108091005804 Peptidases Proteins 0.000 title abstract description 51
- 239000004365 Protease Substances 0.000 title abstract description 41
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 title abstract 5
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 40
- 108090000787 Subtilisin Proteins 0.000 claims abstract description 27
- 239000000203 mixture Substances 0.000 claims description 43
- 238000006073 displacement reaction Methods 0.000 claims description 31
- 241000193422 Bacillus lentus Species 0.000 claims description 20
- 150000001413 amino acids Chemical class 0.000 claims description 16
- 238000004140 cleaning Methods 0.000 claims description 13
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 9
- 239000013604 expression vector Substances 0.000 claims description 5
- 239000004753 textile Substances 0.000 claims description 3
- 102000035195 Peptidases Human genes 0.000 abstract description 39
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 30
- 125000000539 amino acid group Chemical group 0.000 abstract description 30
- 239000002243 precursor Substances 0.000 abstract description 30
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract description 9
- 238000012986 modification Methods 0.000 abstract description 5
- 230000004048 modification Effects 0.000 abstract description 5
- 238000006467 substitution reaction Methods 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract 1
- 102200025035 rs786203989 Human genes 0.000 description 420
- 102200118280 rs33918343 Human genes 0.000 description 410
- 102000004190 Enzymes Human genes 0.000 description 63
- 108090000790 Enzymes Proteins 0.000 description 63
- 108010056079 Subtilisins Proteins 0.000 description 63
- 102000005158 Subtilisins Human genes 0.000 description 63
- 239000003599 detergent Substances 0.000 description 55
- 102220245308 rs1555601006 Human genes 0.000 description 55
- 102220499813 Carbonic anhydrase 2_N62D_mutation Human genes 0.000 description 52
- 235000019419 proteases Nutrition 0.000 description 33
- 229940088598 enzyme Drugs 0.000 description 30
- 108090000623 proteins and genes Proteins 0.000 description 27
- 102200029981 rs28936700 Human genes 0.000 description 25
- 238000000034 method Methods 0.000 description 21
- 235000001014 amino acid Nutrition 0.000 description 19
- 229940024606 amino acid Drugs 0.000 description 18
- 239000002002 slurry Substances 0.000 description 16
- 230000000875 corresponding effect Effects 0.000 description 13
- 210000004027 cell Anatomy 0.000 description 12
- 108020004414 DNA Proteins 0.000 description 11
- 102220554731 Hepatocyte nuclear factor 1-alpha_G20R_mutation Human genes 0.000 description 11
- 102000012479 Serine Proteases Human genes 0.000 description 11
- 108010022999 Serine Proteases Proteins 0.000 description 11
- 102220351005 c.143C>T Human genes 0.000 description 11
- 230000008859 change Effects 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 9
- 102220246481 rs147782267 Human genes 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 8
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 241000194108 Bacillus licheniformis Species 0.000 description 6
- 102000004157 Hydrolases Human genes 0.000 description 6
- 108090000604 Hydrolases Proteins 0.000 description 6
- 238000006555 catalytic reaction Methods 0.000 description 6
- 230000002797 proteolythic effect Effects 0.000 description 6
- 102220072708 rs138992849 Human genes 0.000 description 6
- 102220042823 rs35942089 Human genes 0.000 description 6
- 102200027868 rs62516151 Human genes 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- 244000063299 Bacillus subtilis Species 0.000 description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 description 5
- 102220553631 CBY1-interacting BAR domain-containing protein 1_K27N_mutation Human genes 0.000 description 5
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 5
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 5
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 5
- 239000000654 additive Substances 0.000 description 5
- 230000008034 disappearance Effects 0.000 description 5
- 238000003780 insertion Methods 0.000 description 5
- 230000037431 insertion Effects 0.000 description 5
- 230000014616 translation Effects 0.000 description 5
- 230000000996 additive effect Effects 0.000 description 4
- 102220415048 c.113C>G Human genes 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 102220224495 rs1060503668 Human genes 0.000 description 4
- 102220283107 rs1218848193 Human genes 0.000 description 4
- 102220036452 rs137882485 Human genes 0.000 description 4
- 102220316629 rs1553619289 Human genes 0.000 description 4
- 102200093472 rs1800574 Human genes 0.000 description 4
- 102200024254 rs864309502 Human genes 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 238000013519 translation Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 102220499807 Carbonic anhydrase 2_N62H_mutation Human genes 0.000 description 3
- 108090000317 Chymotrypsin Proteins 0.000 description 3
- 102220493167 FGGY carbohydrate kinase domain-containing protein_N43K_mutation Human genes 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 108091034117 Oligonucleotide Proteins 0.000 description 3
- 102220569900 Phosphoenolpyruvate carboxykinase [GTP], mitochondrial_E89Q_mutation Human genes 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 3
- 102220583535 Small nuclear ribonucleoprotein F_T58S_mutation Human genes 0.000 description 3
- 102220513367 Synaptotagmin-8_H17Q_mutation Human genes 0.000 description 3
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 102220346128 c.59G>T Human genes 0.000 description 3
- 102220350531 c.80A>G Human genes 0.000 description 3
- 229960002376 chymotrypsin Drugs 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 239000006260 foam Substances 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102220196582 rs1057518943 Human genes 0.000 description 3
- 102220248108 rs377129682 Human genes 0.000 description 3
- 102220313493 rs746811389 Human genes 0.000 description 3
- 102220093346 rs876661018 Human genes 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 102220495819 Actin-like protein 6A_H17L_mutation Human genes 0.000 description 2
- 241000228212 Aspergillus Species 0.000 description 2
- 102220581852 Caspase recruitment domain-containing protein 16_S57P_mutation Human genes 0.000 description 2
- 241000206602 Eukaryota Species 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- 102220484443 Myb/SANT-like DNA-binding domain-containing protein 1_L42V_mutation Human genes 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 101710118538 Protease Proteins 0.000 description 2
- 102100023152 Scinderin Human genes 0.000 description 2
- 101710135785 Subtilisin-like protease Proteins 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- 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 2
- 238000010364 biochemical engineering Methods 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 108010043393 protease N Proteins 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 102220132395 rs1057515456 Human genes 0.000 description 2
- 102220225966 rs1064793727 Human genes 0.000 description 2
- 102220228121 rs1064795729 Human genes 0.000 description 2
- 102200115358 rs121918322 Human genes 0.000 description 2
- 102220283756 rs1300016035 Human genes 0.000 description 2
- 102220219210 rs142454490 Human genes 0.000 description 2
- 102220061994 rs144792613 Human genes 0.000 description 2
- 102220264941 rs1457110544 Human genes 0.000 description 2
- 102220341170 rs199565868 Human genes 0.000 description 2
- 102200133203 rs3218619 Human genes 0.000 description 2
- 102200129657 rs35278779 Human genes 0.000 description 2
- 102220261561 rs374050750 Human genes 0.000 description 2
- 102220082985 rs375349172 Human genes 0.000 description 2
- 102200089607 rs398123481 Human genes 0.000 description 2
- 102200047002 rs61752065 Human genes 0.000 description 2
- 102220276580 rs752209909 Human genes 0.000 description 2
- 102220087975 rs777340009 Human genes 0.000 description 2
- 102200091604 rs863225072 Human genes 0.000 description 2
- 108010073419 scinderin Proteins 0.000 description 2
- 239000000344 soap Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 108091005508 Acid proteases Proteins 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 102220582865 Cellular tumor antigen p53_S37T_mutation Human genes 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 102000018361 Contactin Human genes 0.000 description 1
- 108060003955 Contactin Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 108010062466 Enzyme Precursors Proteins 0.000 description 1
- 102000010911 Enzyme Precursors Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 1
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 108010006035 Metalloproteases Proteins 0.000 description 1
- 102000005741 Metalloproteases Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000192041 Micrococcus Species 0.000 description 1
- 241000191938 Micrococcus luteus Species 0.000 description 1
- 108010047320 Pepsinogen A Proteins 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 101710180958 Putative aminoacrylate hydrolase RutD Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241000630486 Robertus Species 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 description 1
- 241000187392 Streptomyces griseus Species 0.000 description 1
- 108700005078 Synthetic Genes Proteins 0.000 description 1
- 101710097834 Thiol protease Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000004442 acylamino group Chemical group 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 125000000266 alpha-aminoacyl group Chemical group 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 235000019730 animal feed additive Nutrition 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000012219 cassette mutagenesis Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000002447 crystallographic data Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- AJIPIJNNOJSSQC-NYLIRDPKSA-N estetrol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H]([C@H](O)[C@@H]4O)O)[C@@H]4[C@@H]3CCC2=C1 AJIPIJNNOJSSQC-NYLIRDPKSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 108010020132 microbial serine proteinases Proteins 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000000869 mutational effect Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 235000019833 protease Nutrition 0.000 description 1
- 230000005892 protein maturation Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 102200093155 rs104894030 Human genes 0.000 description 1
- 102220004396 rs121964855 Human genes 0.000 description 1
- 102220267892 rs1555193020 Human genes 0.000 description 1
- 102220215697 rs200971068 Human genes 0.000 description 1
- 102220041795 rs587780747 Human genes 0.000 description 1
- 102220279241 rs587780812 Human genes 0.000 description 1
- 102220254284 rs755928199 Human genes 0.000 description 1
- 102220103399 rs878854207 Human genes 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 108010059841 serine carboxypeptidase Proteins 0.000 description 1
- 239000002453 shampoo Substances 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38681—Chemically modified or immobilised enzymes
-
- 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)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
- C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G4/00—Chewing gum
- A23G4/06—Chewing gum characterised by the composition containing organic or inorganic compounds
- A23G4/12—Chewing gum characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
- A23G4/123—Chewing gum characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/66—Enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/10—Washing or bathing preparations
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38609—Protease or amylase in solid compositions only
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38618—Protease or amylase in liquid compositions only
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/39—Organic or inorganic per-compounds
- C11D3/3902—Organic or inorganic per-compounds combined with specific additives
- C11D3/3905—Bleach activators or bleach catalysts
- C11D3/3907—Organic compounds
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/39—Organic or inorganic per-compounds
- C11D3/3945—Organic per-compounds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21062—Subtilisin (3.4.21.62)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/86—Products or compounds obtained by genetic engineering
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
- C11D2111/10—Objects to be cleaned
- C11D2111/12—Soft surfaces, e.g. textile
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D2111/00—Cleaning compositions characterised by the objects to be cleaned; Cleaning compositions characterised by non-standard cleaning or washing processes
- C11D2111/10—Objects to be cleaned
- C11D2111/14—Hard surfaces
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Veterinary Medicine (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- General Engineering & Computer Science (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Animal Husbandry (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Nutrition Science (AREA)
- Detergent Compositions (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Cosmetics (AREA)
- Slot Machines And Peripheral Devices (AREA)
- Exhaust Gas After Treatment (AREA)
- Finish Polishing, Edge Sharpening, And Grinding By Specific Grinding Devices (AREA)
Abstract
公开了由天然存在的或重组的非人类蛋白酶的DNA序列衍生的新型蛋白酶变体。变异蛋白酶的获得方法通常是,对编码天然存在的或重组的蛋白酶的前体DNA序列进行体外修饰,在前体蛋白酶的氨基酸序列中引起多处氨基酸残基的置换。这种变体蛋白酶具有与前体蛋白酶不同的性质,如改变的洗涤性能。置换的氨基酸残基相应于解淀粉芽孢杆菌枯草杆菌蛋白酶位置62、212、230、232、252和257。
Description
相关申请
本申请是1998年10月23日提交的美国专利申请08/956,323、1998年10月23日提交的美国专利申请08/956,564和1998年10月23日提交的美国专利申请08/956,324的部分继续申请，所有这些申请在此都完整引用。
发明背景
丝氨酸蛋白酶是羰基水解酶的一个亚组。其包括多种类别的具有广泛特性和生物功能的酶。Stroud,R.,科学美国人(Sci.Amer.)131:74-88。尽管其功能的多样性，但丝氨酸蛋白酶的催化机制至少由两种遗传分离的酶家族来完成：1)枯草杆菌蛋白酶和2)哺乳动物胰凝乳蛋白酶相关的丝氨酸蛋白酶及同源细菌丝氨酸蛋白酶(例如胰蛋白酶和灰色链霉菌(S.gresius)胰蛋白酶)。这两个丝氨酸蛋白酶家族显示非常相似的催化机理。Kraut,J.(1977)，生物化学年述(Annu.Rev.Biochem.),46:331-358。此外，尽管一级结构不相关，但这两个酶家族的三级结构存在由丝氨酸、组氨酸和天冬氨酸组成的氨基酸保守催化三联体。
枯草杆菌蛋白酶是由多种芽孢杆菌属的种及其它微生物大量分泌的丝氨酸蛋白酶(分子量约为27,500)。已经测定了芽孢杆菌属至少九个不同种的枯草杆菌蛋白酶的蛋白质序列。Markland,F.S.等人(1983),Hooppe-Seyler’s Z.Physiol.Chem.,364:1537-1540。曾经报道了来源于解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、地衣芽孢杆菌(Bacilluslicheniformis)和迟缓芽孢杆菌(B.lentus)几种天然变体的枯草杆菌蛋白酶的三维晶体结构。这些研究表明，尽管枯草杆菌蛋白酶与哺乳动物丝氨酸蛋白酶在遗传上是不相关的，但它具有相似的活性部位结构。含有共价结合的肽抑制剂的枯草杆菌蛋白酶(Robertus,J.D.等人(1972)，生物化学(Biochemistry),11:2439-2449)或产物复合物(Robertus,J.D.等人(1976)，生物化学杂志(J.Biol.Chem.),251:1097-1103)的x-射线晶体结构也提供了关于枯草杆菌蛋白酶的活性部位和推断的底物结合裂隙的信息。另外，曾经报道了关于枯草杆菌蛋白酶的大量动力学和化学修饰研究(Svendsen,B.(1976),Carlsberg研究通讯(Carlsberg Res.Commun.),41:237-291；Markland,F.S.,同前)，并且至少有一条报道，其中枯草杆菌蛋白酶残基222的甲硫氨酸的侧链被过氧化氢转化为甲硫氨酸-亚砜(Stauffer,D.C.等人(1965)，生物化学杂志，244:5333-5338)，并进行了广泛的位点专一诱变(Wells和Estell(1988)TIBS 13:291-297)。
发明概述
本发明的一个目的在于提供在一个或多个残基位置处含有氨基酸置换的蛋白酶变体，残基位置相应于选自解淀粉芽孢杆菌枯草杆菌蛋白酶位置62、212、230、232、252和257的残基。
尽管可利用任何上述所列氨基酸置换的任一组合，本发明优选的蛋白酶变体酶包括如下组合的氨基酸残基的置换。所有的残基位置相应于解淀粉芽孢杆菌枯草杆菌蛋白酶位置：
(1)一种蛋白酶变体，包括在位置62以及在一个或多个如下位置：
103、104、109、159、213、232、236、245、248和252中的
氨基酸残基的置换；
(2)一种蛋白酶变体，包括在位置212以及在一个或多个如下位
置：12、98、102、103、104、159、232、236、245、248和252
中的氨基酸残基的置换；
(3)一种蛋白酶变体，包括在位置230以及在一个或多个如下位置：
68、103、104、159、232、236、和245中的氨基酸残基的置
换；
(4)一种蛋白酶变体，包括在位置232以及在一个或多个如下位置：
1、9、12、61、62、68、76、97、98、101、102、103、104、
109、130、131、159、183、185、205、209、210、212、213、
217、230、236、245、248、252、257、260、270和275中的
氨基酸残基的置换；
(5)一种蛋白酶变体，包括在位置232以及在一个或多个如下位置：
103、104、236和245中的氨基酸残基的置换；
(6)一种蛋白酶变体，包括在位置232和103以及在一个或多个如
下位置：1、9、12、61、62、68、76、97、98、101、102、103、
104、109、130、131、159、183、185、205、209、210、212、
213、217、230、236、245、248、252、257、260、270和275
中的氨基酸残基的置换；
(7)一种蛋白酶变体，包括在位置232和104以及在一个或多个如
下位置：1、9、12、61、62、68、76、97、98、101、102、103、
104、109、130、131、159、183、185、205、209、210、212、
213、217、230、236、245、248、252、257、260、270和275
中的氨基酸残基的置换；
(8)一种蛋白酶变体，包括在位置232和236以及在一个或多个如
下位置：1、9、12、61、62、68、76、97、98、101、102、103、
104、109、130、131、159、183、185、205、209、210、212、
213、217、230、236、245、248、252、257、260、270和275
中的氨基酸残基的置换；
(9)一种蛋白酶变体，包括在位置232和245以及在一个或多个如
下位置：1、9、12、61、62、68、76、97、98、101、102、103、
104、109、130、131、159、183、185、205、209、210、212、
213、217、230、236、245、248、252、257、260、270和275
中的氨基酸残基的置换；
(10)一种蛋白酶变体，包括在位置232、103、104、236和245
以及在一个或多个如下位置：1、9、12、61、62、68、76、97、
98、101、102、103、104、109、130、131、159、183、185、
205、209、210、212、213、217、230、236、245、248、252、
257、260、270和275中的氨基酸残基的置换；
(11)一种蛋白酶变体，包括在位置252以及在一个或多个如下位
置：1、9、12、61、62、68、76、97、98、101、102、103、104、
109、130、131、159、183、185、210、212、213、217、236、
245、248、和270中的氨基酸残基的置换；
(12)一种蛋白酶变体，包括在位置252以及在一个或多个如下位
置：103、104、236和245中的氨基酸残基的置换；
(13)一种蛋白酶变体，包括在位置252和103以及在一个或多个
如下位置：1、9、12、61、62、68、97、98、101、102、103、
104、109、130、131、159、183、185、210、212、213、217、
232、236、245、248、和270中的氨基酸残基的置换；
(14)一种蛋白酶变体，包括在位置252和104以及在一个或多个
如下位置：1、9、12、61、62、68、97、98、101、102、103、
104、109、130、131、159、183、185、210、212、213、217、
232、236、245、248和270中的氨基酸残基的置换；
(15)一种蛋白酶变体，包括在位置252和236以及在一个或多个
如下位置：1、9、12、61、62、68、97、98、101、102、103、
104、109、130、131、159、183、185、210、212、213、217、
232、236、245、248、和270中的氨基酸残基的置换；
(16)一种蛋白酶变体，包括在位置252和245以及在一个或多个
如下位置：1、9、12、61、62、68、97、98、101、102、103、
104、109、130、131、159、183、185、210、212、213、217、
232、236、245、248和270中的氨基酸残基的置换；
(17)一种蛋白酶变体，包括在位置252、103、104、236和245
以及在一个或多个如下位置：1、9、12、61、62、68、97、98、
101、102、103、104、109、130、131、159、183、185、210、
212、213、217、232、236、245、248和270中的氨基酸残基
的置换；和
(18)一种蛋白酶变体，包括在位置257以及在一个或多个如下位
置：68、103、104、205、209、210、232、236、245、270和
275中的氨基酸残基的置换。
更优选的蛋白酶变体的置换选自相应于解淀粉芽孢杆菌枯草杆菌蛋白酶的表1中位置的残基位置。
表1
| 76 | 103 | 104 | 212 | 271 | ||||||||
| 76 | 103 | 104 | 252 | 261 | ||||||||
| 76 | 103 | 104 | 212 | 258 | ||||||||
| 4 | 76 | 103 | 104 | 159 | 217 | 252 | ||||||
| 12 | 62 | 76 | 103 | 104 | 159 | |||||||
| 76 | 103 | 104 | 212 | 268 | 271 | |||||||
| 76 | 87 | 103 | 104 | 212 | 271 | |||||||
| 76 | 103 | 104 | 212 | 245 | 271 | |||||||
| 76 | 103 | 104 | 134 | 141 | 212 | 271 | ||||||
| 76 | 103 | 104 | 212 | 236 | 243 | 271 | ||||||
| 20 | 62 | 76 | 103 | 104 | ||||||||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | |||||
| 76 | 103 | 104 | 232 | 245 | ||||||||
| 24 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | |||||
| 68 | 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | ||||||
| 68 | 103 | 104 | 140 | 159 | 232 | 236 | 245 | 252 |
表1(续)
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | ||||
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | |||||
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | ||||
| 68 | 87 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | 275 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | |||||
| 68 | 103 | 104 | 116 | 159 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | |||||
| 10 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 203 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 232 | 236 | 237 | 245 | |||||
| 68 | 76 | 79 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 68 | 103 | 104 | 159 | 183 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 174 | 206 | 232 | 236 | 245 | ||||
| 68 | 103 | 104 | 159 | 188 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 230 | 232 | 236 | 245 | |||||
| 68 | 98 | 103 | 104 | 159 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 215 | 232 | 236 | 245 | |||||
| 68 | 76 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | ||||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | ||||
| 76 | 103 | 104 | 232 | 236 | 245 | 257 | ||||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | 275 | ||||
| 76 | 103 | 104 | 257 | 275 | ||||||||
| 68 | 103 | 104 | 159 | 224 | 232 | 236 | 245 | 257 | ||||
| 76 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | |||||
| 68 | 76 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | ||||
| 68 | 76 | 103 | 104 | 159 | 211 | 232 | 236 | 245 | ||||
| 12 | 68 | 76 | 103 | 104 | 159 | 214 | 232 | 236 | 245 | |||
| 68 | 76 | 103 | 104 | 159 | 215 | 232 | 236 | 245 | ||||
| 12 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 20 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 259 | |||
| 68 | 87 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 260 | |||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 261 | ||||
| 76 | 103 | 104 | 232 | 236 | 242 | 245 | ||||||
| 68 | 76 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | ||||
| 12 | 48 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | |||
| 76 | 103 | 104 | 232 | 236 | 245 |
表1(续)
| 76 | 103 | 104 | 159 | 192 | 232 | 236 | 245 | |||||
| 76 | 103 | 104 | 147 | 159 | 232 | 236 | 245 | 248 | 251 | |||
| 12 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 272 | |||
| 68 | 76 | 103 | 104 | 159 | 183 | 206 | 232 | 236 | 245 | |||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 256 | ||||
| 68 | 76 | 103 | 104 | 159 | 206 | 232 | 236 | 245 | ||||
| 27 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 68 | 76 | 103 | 104 | 116 | 159 | 170 | 185 | 232 | 236 | 245 | ||
| 61 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 99 | 159 | 184 | 232 | 236 | 245 | 248 | 252 | ||
| 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 109 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 20 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 261 | |||
| 68 | 103 | 104 | 159 | 185 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 185 | 210 | 232 | 236 | 245 | 248 | 252 | ||
| 68 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 213 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 103 | 104 | 159 | 215 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 216 | 232 | 236 | 245 | 248 | 252 | |||
| 20 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 173 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 251 | 252 | |||
| 68 | 103 | 104 | 159 | 206 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 55 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 255 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 256 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 260 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 257 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 258 | |||
| 8 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 269 |
表1(续)
| 68 | 103 | 104 | 116 | 159 | 232 | 236 | 245 | 248 | 252 | 260 | ||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 261 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 261 | |||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 232 | 236 | 245 | 248 | 252 | |||||
| 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 18 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 76 | 101 | 103 | 104 | 159 | 213 | 218 | 232 | 236 | 245 | 260 | |
| 68 | 103 | 104 | 159 | 228 | 232 | 236 | 245 | 248 | 252 | |||
| 33 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||
| 68 | 76 | 89 | 103 | 104 | 159 | 210 | 213 | 232 | 236 | 245 | 260 | |
| 61 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||
| 103 | 104 | 159 | 205 | 210 | 232 | 236 | 245 | |||||
| 61 | 68 | 103 | 104 | 130 | 159 | 232 | 236 | 245 | 248 | 252 | ||
| 61 | 68 | 103 | 104 | 133 | 137 | 159 | 232 | 236 | 245 | 248 | 252 | |
| 61 | 103 | 104 | 133 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 103 | 104 | 159 | 218 | 232 | 236 | 245 | 248 | 252 | |||
| 61 | 68 | 103 | 104 | 159 | 160 | 232 | 236 | 245 | 248 | 252 | ||
| 3 | 61 | 68 | 76 | 103 | 104 | 232 | 236 | 245 | 248 | 252 | ||
| 61 | 68 | 103 | 104 | 159 | 167 | 232 | 236 | 245 | 248 | 252 | ||
| 97 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 98 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 99 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 101 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 102 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 106 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 109 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 261 | ||||
| 62 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 184 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 166 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 217 | 232 | 236 | 245 | 248 | 252 | ||||
| 20 | 62 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 62 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | |||
| 103 | 104 | 159 | 206 | 217 | 232 | 236 | 245 | 248 | 252 |
表1(续)
| 62 | 103 | 104 | 159 | 206 | 232 | 236 | 245 | 248 | 252 | |||
| 103 | 104 | 130 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 131 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 27 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 38 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 38 | 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | |||
| 68 | 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | 271 | ||
| 68 | 76 | 103 | 104 | 159 | 209 | 213 | 232 | 236 | 245 | 260 | ||
| 68 | 76 | 103 | 104 | 159 | 210 | 213 | 232 | 236 | 245 | 260 | ||
| 68 | 76 | 103 | 104 | 159 | 205 | 213 | 232 | 236 | 245 | 260 | ||
| 68 | 76 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | 260 | |||
| 68 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | ||||
| 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | ||||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 230 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 126 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 205 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | |||||
| 103 | 104 | 159 | 230 | 236 | 245 | |||||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 260 | |||||
| 103 | 104 | 159 | 232 | 236 | 245 | |||||||
| 68 | 103 | 104 | 159 | 174 | 232 | 236 | 245 | 257 | ||||
| 68 | 103 | 104 | 159 | 194 | 232 | 236 | 245 | 257 | ||||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 257 | ||||
| 103 | 104 | 159 | 232 | 236 | 245 | 257 | ||||||
| 68 | 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | 261 | ||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | 261 | ||||
| 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | |||||
| 103 | 104 | 159 | 210 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 209 | 232 | 236 | 245 | 257 | |||||
| 68 | 76 | 103 | 104 | 159 | 210 | 213 | 232 | 236 | 245 | 260 | ||
| 12 | 103 | 104 | 159 | 209 | 213 | 232 | 236 | 245 | 260 | |||
| 103 | 104 | 209 | 232 | 236 | 245 | 257 | ||||||
| 103 | 104 | 159 | 205 | 210 | 213 | 232 | 236 | 245 | 260 | |||
| 103 | 104 | 159 | 205 | 209 | 232 | 236 | 245 | 260 | ||||
| 68 | 103 | 104 | 159 | 205 | 209 | 210 | 232 | 236 | 245 | |||
| 103 | 104 | 159 | 205 | 209 | 210 | 232 | 236 | 245 | 257 |
表1(续)
| 103 | 104 | 159 | 205 | 209 | 232 | 236 | 245 | 257 | ||||
| 68 | 103 | 104 | 159 | 205 | 209 | 210 | 232 | 236 | 245 | 260 | ||
| 103 | 104 | 159 | 205 | 209 | 210 | 232 | 236 | 245 | ||||
| 103 | 104 | 159 | 209 | 210 | 232 | 236 | 245 | |||||
| 103 | 104 | 159 | 205 | 210 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 128 | 159 | 232 | 236 | 245 | |||||
| 48 | 103 | 104 | 159 | 230 | 236 | 245 | ||||||
| 48 | 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | ||||
| 48 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 48 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | 261 | |||
| 102 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 12 | 102 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | ||
| 101 | 102 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | ||
| 98 | 102 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | ||
| 102 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | |||
| 103 | 104 | 131 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 184 | 232 | 236 | 245 | 248 | 252 | ||||
| 103 | 104 | 159 | 232 | 236 | 244 | 245 | 248 | 252 | ||||
| 62 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | 256 | ||
| 12 | 62 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 101 | 103 | 104 | 159 | 185 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 206 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | |||
| 98 | 102 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 102 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 98 | 102 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | ||
| 98 | 102 | 103 | 104 | 159 | 212 | 232 | 236 | 248 | 252 | |||
| 62 | 103 | 104 | 109 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 62 | 103 | 104 | 159 | 212 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 62 | 101 | 103 | 104 | 159 | 212 | 213 | 232 | 236 | 245 | 248 | 252 | |
| 103 | 104 | 159 | 232 | 245 | 248 | 252 | ||||||
| 103 | 104 | 159 | 230 | 245 | ||||||||
| 62 | 103 | 104 | 130 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 101 | 103 | 104 | 130 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 128 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 62 | 101 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 62 | 103 | 104 | 128 | 159 | 213 | 232 | 236 | 245 | 248 | 252 | ||
| 62 | 103 | 104 | 128 | 159 | 213 | 232 | 236 | 245 | 248 | 252 |
表1(续)
| 101 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 260 | |||
| 101 | 103 | 104 | 131 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 98 | 101 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 99 | 101 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 205 | 232 | 236 | 245 | 248 | 252 | |||
| 101 | 103 | 104 | 159 | 230 | 236 | 245 | ||||||
| 101 | 103 | 104 | 159 | 194 | 232 | 236 | 245 | 248 | 252 | |||
| 76 | 101 | 103 | 104 | 159 | 194 | 232 | 236 | 245 | 248 | 252 | ||
| 101 | 103 | 104 | 159 | 230 | 232 | 236 | 245 | 248 | 252 | |||
| 62 | 103 | 104 | 159 | 185 | 206 | 213 | 232 | 236 | 245 | 248 | 252 | 271 |
更优选的蛋白酶变体的置换选自相应于解淀粉芽孢杆菌枯草杆菌蛋白酶的表2中位置的残基位置。
表2
| N76D | S103A | V104I | S212P | E271V | ||||||||
| N76D | S103A | V104I | N252K | N261Y | ||||||||
| N76D | S103A | V104I | S212P | G258R | ||||||||
| V4E | N76D | S103A | V104I | G159D | L217E | N252D | ||||||
| Q12H | N62H | N76D | S103A | V104I | G159D | |||||||
| N76D | S103A | V104I | S212P | V268F | E271V | |||||||
| N76D | S87R | S103A | V104I | S212P | E271V | |||||||
| N76D | S103A | V104I | S212P | Q245L | E271V | |||||||
| N76D | S103A | V104I | T134S | S141N | S212P | E271V | ||||||
| N76D | S103A | V104I | S212P | Q236L | N243S | E271V | ||||||
| G20V | N62S | N76D | S103A | V104I | ||||||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| N76D | S103A | V104I | A232V | Q245R | ||||||||
| S24T | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | |||||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||||
| V68A | S103A | V104I | N140D | G159D | A232V | Q236H | Q245R | N252K |
表2(续)
| N43S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | ||||
| N43K | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | ||||
| V68A | S87G | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | R275S | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | |||||
| V68A | S103A | V104I | N116D | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | |||||
| R10C | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | V203E | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | K237E | Q245R | |||||
| V68A | N76D | 179N | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | N183D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A174V | Q206L | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | S188C | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A230T | A232V | Q236H | Q245R | |||||
| V68A | A98T | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A215T | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248S | |||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R |
表2(续)
| V68A | N76D | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | ||||
| N76D | S103A | V104I | A232V | Q236H | Q245R | L257V | ||||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | R275H | ||||
| N76D | S103A | V104I | L257V | R275H | ||||||||
| V68A | S103A | V104I | G159D | T224A | A232V | Q236H | Q245R | L257V | ||||
| N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | |||||
| V68A | N76D | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | G211R | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | G211V | A232V | Q236H | Q245R | ||||
| Q12R | V68A | N76D | S103A | V104I | G159D | Y214L | A232V | Q236H | Q245R | |||
| V68A | N76D | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | ||||
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| G20R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S259G | |||
| V68A | S87R | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | T260V | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261G | ||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261W | ||||
| N76D | S103A | V104I | A232V | Q236H | S242P | Q245R | ||||||
| V68A | N76D | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R |
表2(续)
| Q12R | A48V | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | |||
| N76D | S103A | V104I | A232V | Q236H | Q245R | |||||||
| N76D | S103A | V104I | G159D | Y192F | A232V | Q236H | Q245R | |||||
| N76D | S103A | V104I | V147I | G159D | A232V | Q236H | Q245R | N248S | K251R | |||
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | A272S | |||
| V68A | N76D | S103A | V104I | G159D | N183K | Q206L | A232V | Q236H | Q245R | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S256R | ||||
| V68A | N76D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | ||||
| K27R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | N116T | G159D | R170S | N185S | A232V | Q236H | Q245R | ||
| G61E | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212P | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | S99N | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | ||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| G20R | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | Y209F | A232V | Q236H | Q245R | N248D | N252K |
表2(续)
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261D | |||
| V68A | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210T | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210S | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | N185D | P210L | A232V | Q236H | Q245R | N248D | N252K | ||
| V68A | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212A | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | T213S | A232V | Q236H | Q245R | N248D | N252K | ||||
| V68A | A103V | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213G | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A215V | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S216T | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S216V | A232V | Q236H | Q245R | N248D | N252K |
表2(续)
| V68A | S103A | V104I | G159D | S216C | A232V | Q236H | Q245R | N248D | N252K | |||
| G20A | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | N173D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | K251V | N252K | |||
| V68A | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252L | ||||
| P55S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T255V | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256N | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256E | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T260R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | L257R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | G258D | |||
| I8V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N269D | ||
| V68A | S103A | V104I | N116S | G159D | A232V | Q236H | Q245R | N248D | N252K | T260E | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261D |
表2(续)
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | |||||
| S103A | V104I | G159D | A232S | Q236H | Q245R | N248D | N252K | |||||
| V68A | S103A | V104I | G159D | A232V | Q236R | Q245R | N248D | N252K | ||||
| N18S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245V | N248D | N252K | ||||
| V68A | N76D | S101T | S103A | V104I | G159D | T213R | N218S | A232V | Q236H | Q245R | T260A | |
| V68A | S103A | V104I | G159D | A228V | A232V | Q236H | Q245R | N248D | N252K | |||
| T33S | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| V68A | N76D | E89D | S103A | V104I | G159D | P210L | T213R | A232V | Q236H | Q245R | T260A | |
| G61E | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| S103A | V104I | G159D | V205I | P210I | A232V | Q236H | Q245R | |||||
| G61E | V68A | S103A | V104I | S130A | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| G61E | V68A | S103A | V104I | A133S | Q137R | G159D | A232V | Q236H | Q245R | N248D | N252K | |
| G61E | S103A | V104I | A133V | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248G | N252K | ||||
| V68A | S103A | V104I | G159D | N218S | A232V | Q236H | Q245R | N248D | N252K | |||
| G61E | V68A | S103A | V104I | G159D | S160V | A232V | Q236H | Q245R | N248D | N252K | ||
| S3L | G61E | V68A | N76D | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K |
表2(续)
| G61E | V68A | S103A | V104I | G159D | S167F | A232V | Q236H | Q245R | N248D | N252K | ||
| G97E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| A98D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S99E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S101E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | S106E | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | Q109E | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | ||||
| S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| N62D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | S166D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | L217E | A232V | Q236H | Q245R | N248D | N252K | ||||
| G20R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| S103A | V104I | G159D | Q206R | L217E | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K |
表2(续)
| S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| K27N | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| T38G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| T38A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | |||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | E271G | ||
| V68A | N76D | S103A | V104I | G159D | Y209W | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | P210I | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | V205I | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | T260A | |||
| V68A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | ||||
| N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | ||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A230V | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104 I | G159D | L126F | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | |||||
| S103A | V104I | G159D | A230V | Q236H | Q245R |
表2(续)
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | T260A | |||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | |||||||
| V68A | S103A | V104I | G159D | A174V | A232V | Q236H | Q245R | L257V | ||||
| V68A | S103A | V104I | G159D | A194S | A232V | Q236H | Q245R | L257V | ||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | L257V | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | ||||||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | N261W | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | N261W | ||||
| S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | |||||
| S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | L257V | |||||
| V68A | N76D | S103A | V104I | G159D | P210L | T213R | A232V | Q236H | Q245R | T260A | ||
| Q12R | S103A | V104I | G159D | Y209W | T213R | A232V | Q236H | Q245R | T260A | |||
| S103A | V104I | Y209W | A232V | Q236H | Q245R | L257V | ||||||
| S103A | V104I | G159D | V205I | P210I | T213R | A232V | Q236H | Q245R | T260A | |||
| S103A | V104I | G159D | V205I | Y209W | A232V | Q236H | Q245R | T260A | ||||
| V68A | S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | |||
| S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | L257V | |||
| S103A | V104I | G159D | V205I | Y209W | A232V | Q236H | Q245R | L257V |
表2(续)
| IV68A | S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | T260A | ||
| S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | ||||
| S103A | V104I | G159D | Y209W | P210I | A232V | Q236H | Q245R | |||||
| S103A | V104I | G159D | V205I | P210I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | |||||
| A48V | S103A | V104I | G159D | A230V | Q236H | Q245R | ||||||
| A48V | V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | ||||
| A48V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A48V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | N261W | |||
| G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | |||
| Q12R | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| G102A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184S | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184G | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | V244T | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | V244A | Q245R | N248D | N252K |
表2(续)
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | S256R | ||
| Q12R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | Q206E | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | T213Q | A232V | Q236H | Q245R | N248D | N252K | |||
| A98L | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | N248D | N252K | |||
| N62D | S103A | V104I | Q109R | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | G159D | S212G | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S101G | S103A | V104I | G159D | S212G | T213R | A232V | Q236H | Q245R | N248D | N252K | |
| S103A | V104I | G159D | A232V | Q245R | N248D | N252K | ||||||
| S103A | V104I | G159D | A230V | Q245R | ||||||||
| N62D | S103A | V104I | S130G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | S128G | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S101G | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K |
| N62D | S103A | V104I | S128G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | S128L | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T260A | |||
| S101G | S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A98V | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S99G | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | A230V | Q236H | Q245R | ||||||
| S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | |||
| N76D | S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | G159D | A230V | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S103A | V104I | G159D | N185D | Q206E | T213R | A232V | Q236H | Q245R | N248D | N252K | E271Q |
本发明另外一个目的在于提供编码这些蛋白酶变体的DNA序列，以及含有这些变异体DNA序列的表达载体。
更进一步，本发明的另一目的在于提供用这些载体转化的宿主细胞，以及能够表达这种DNA而在细胞内或细胞外产生蛋白酶变体的宿主细胞。
进一步提供一种含有本发明的蛋白酶变体的清洁用组合物。
另外，也提供一种含有本发明的蛋白酶变体的动物饲料。
还提供了一种用于处理纺织品的含有本发明的蛋白酶变体之组合物。
附图简述
图1A-C描述解淀粉芽孢杆菌枯草杆菌蛋白酶的DNA及氨基酸序列，和该基因的部分限制酶切图谱。
图2描述解淀粉芽孢杆菌(BPN)’和迟缓芽孢杆菌(野生型)枯草杆菌蛋白酶之间的保守氨基酸残基。
图3A和3B描述四种枯草杆菌蛋白酶的氨基酸序列。最上一行代表来源于解淀粉芽孢杆菌枯草杆菌蛋白酶(有时也称为枯草杆菌蛋白酶BPN’)的枯草杆菌蛋白酶的氨基酸序列。第二行描述来源于枯草芽孢杆菌的枯草杆菌蛋白酶的氨基酸序列。第三行描述来源于地衣芽孢杆菌的枯草杆菌蛋白酶的氨基酸序列。第四行描述来源于迟缓芽孢杆菌的枯草杆菌蛋白酶(在PCT WO89/06276中也称为枯草杆菌蛋白酶309)的氨基酸序列。符号*表示与枯草杆菌蛋白酶BPN’相比特定氨基酸残基缺如。
发明详述
蛋白酶是通常用来切割蛋白质或肽的肽键的羰基水解酶。在此使用时，“蛋白酶”意思是天然存在的蛋白酶或重组蛋白酶。天然存在的蛋白酶包括α-氨酰肽水解酶、肽酰氨基酸水解酶、酰基氨基水解酶、丝氨酸羧基肽酶、金属羧基肽酶、巯基蛋白酶、羧基蛋白酶和金属蛋白酶。包括丝氨酸、金属、巯基和酸性蛋白酶，以及内切蛋白酶和外切蛋白酶。
本发明包括非天然存在的羰基水解酶变体的蛋白酶(蛋白酶变体)，与衍生出该变体氨基酸序列的前体羰基水解酶相比，其具有不同的蛋白水解活性、稳定性、底物特异性、pH分布曲线和/或性能特征。特别是，这些蛋白酶变体具有自然界中未知的氨基酸序列，它是由于前体蛋白酶的大多数氨基酸残基被替换为不同氨基酸而产生的。前体蛋白酶可以是天然存在的蛋白酶或重组蛋白酶。
此处有用的蛋白酶变体包含19种天然存在的L-氨基酸中任一种在指定氨基酸残基位置处的置换。这些置换能在任何前体枯草杆菌蛋白酶(原核、真核、哺乳动物等)中进行。在整个申请中，以常用的单字母及三字母密码的形式提及不同的氨基酸。这些密码由Dale,M.W.(1989)，《细菌分子遗传学》(Molecular Genetics of Bacteria),John Wiley&Sons,Ltd.,附录B确定。
此处有用的蛋白酶变体优选地来源于芽孢杆菌属的枯草杆菌蛋白酶。更优选地，蛋白酶变体来源于迟缓芽孢杆菌枯草杆菌蛋白酶和/或枯草杆菌蛋白酶309。
枯草杆菌蛋白酶是一般用来切割蛋白质或肽的肽键的细菌或真菌蛋白酶。在此使用时，“枯草杆菌蛋白酶”意思是天然存在的枯草杆菌蛋白酶或重组枯草杆菌蛋白酶。已知一系列天然存在的枯草杆菌蛋白酶是由不同微生物种产生且常常为分泌的。该系列成员的氨基酸序列不是完全同源的。然而，该系列中的枯草杆菌蛋白酶显示相同或相似类型的蛋白水解活性。该类丝氨酸蛋白酶具有一种共同的氨基酸序列，该序列定义了有别于胰凝乳蛋白酶相关种类丝氨酸蛋白酶的催化三联体。枯草杆菌蛋白酶和胰凝乳蛋白酶相关的丝氨酸蛋白酶都含有包括天冬氨酸、组氨酸和丝氨酸的催化三联体。在枯草杆菌蛋白酶相关蛋白酶中，这些氨基酸从氨基端向羧基端阅读的相对顺序是天冬氨酸-组氨酸-丝氨酸。但是，在胰凝乳蛋白酶相关蛋白酶中相对顺序为组氨酸-天冬氨酸-丝氨酸。因此，枯草杆菌蛋白酶在此是指含有枯草杆菌蛋白酶相关蛋白酶的催化三联体的丝氨酸蛋白酶。实例包括但不限于此处图3中所确定的枯草杆菌蛋白酶。一般而言，并且对于本发明，蛋白酶中氨基酸的编号对应于分配给图1所示成熟解淀粉芽孢杆菌枯草杆菌蛋白酶序列的编号。
“重组枯草杆菌蛋白酶”或“重组蛋白酶”是指一种枯草杆菌蛋白酶或蛋白酶，其中编码枯草杆菌蛋白酶或蛋白酶的DNA序列受到修饰，产生一种变异(或突变)DNA序列，该序列编码天然存在的氨基酸序列中一种或多种氨基酸的置换、缺失或插入。产生这种修饰、并且可与此处公开的方法结合的适当方法包括美国专利RE34,606、美国专利5,204,015和美国专利5,185,258、美国专利5,700,676、美国专利5,801,038和美国专利5,763,257中公开的方法。
“非人类的枯草杆菌蛋白酶”及其编码DNA可以从多种原核及真核生物中获得。原核生物的合适实例包括革兰氏阴性生物如大肠杆菌或假单胞菌(Pseudomonas)、革兰氏阳性菌如微球菌(Micrococcus)或芽孢杆菌。可从中获得枯草杆菌蛋白酶及其基因的真核生物的实例包括酵母如酿酒酵母(Saccharomyces cerevisiae)、真菌如曲霉(Aspergillus)属的种。
“蛋白酶变体”具有由“前体蛋白酶”的氨基酸序列衍生的氨基酸序列。前体蛋白酶包括天然存在的蛋白酶和重组蛋白酶。蛋白酶变体的氨基酸序列通过前体氨基酸序列中一种或多种氨基酸的置换、缺失或插入而由前体蛋白酶氨基酸序列“衍生”。这种修饰是编码前体蛋白酶氨基酸序列的“前体DNA序列”的修饰，而不是前体蛋白酶自身的操作。对前体DNA序列进行这种操作的适当方法包括此处公开的方法，以及本领域技术人员公知的方法(参见，例如，EP0 328299、WO89/06279和已经在此引用的美国专利及申请)。
此处鉴定了在相应于选自解淀粉芽孢杆菌枯草杆菌蛋白酶残基位置62、212、230、232、252和257处的一个和多个残基位置的定点置换。
优选的变体是具有在相应于表1中解淀粉芽孢杆菌枯草杆菌蛋白酶位置的残基位置上置换的组合的那些变体。
更优选的变体是具有在相应于表2中解淀粉芽孢杆菌枯草杆菌蛋白酶位置的残基位置上置换的组合的那些变体。
更为优选的变体是具有在相应于表3中解淀粉芽孢杆菌枯草杆菌蛋白酶位置的残基位置上置换的组合的那些变体。
表3
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | |||||
| 68 | 76 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 260 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | ||||||
| 68 | 103 | 104 | 140 | 159 | 232 | 236 | 245 | 252 | ||||
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 252 | ||||
| 43 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | |||||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | |||||
| 68 | 76 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | ||||
| 68 | 103 | 104 | 159 | 224 | 232 | 236 | 245 | 257 | ||||
| 76 | 103 | 104 | 159 | 232 | 236 | 245 | 257 | |||||
| 68 | 76 | 103 | 104 | 159 | 211 | 232 | 236 | 245 | ||||
| 12 | 68 | 76 | 103 | 104 | 159 | 214 | 232 | 236 | 245 | |||
| 68 | 76 | 103 | 104 | 159 | 215 | 232 | 236 | 245 | ||||
| 12 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 20 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 259 | |||
| 68 | 76 | 87 | 103 | 104 | 159 | 232 | 236 | 245 | 260 | |||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 261 | ||||
| 12 | 48 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | |||
| 76 | 103 | 104 | 159 | 192 | 232 | 236 | 245 | |||||
| 76 | 103 | 104 | 147 | 159 | 232 | 236 | 245 | 248 | 251 | |||
| 12 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 272 | |||
| 68 | 76 | 103 | 104 | 159 | 183 | 206 | 232 | 236 | 245 | |||
| 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | 256 | ||||
| 68 | 76 | 103 | 104 | 159 | 206 | 232 | 236 | 245 | ||||
| 27 | 68 | 76 | 103 | 104 | 159 | 232 | 236 | 245 | ||||
| 68 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 109 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 20 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 209 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 210 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 212 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 213 | 232 | 236 | 245 | 248 | 252 |
表3(续)
| 68 | 103 | 104 | 213 | 232 | 236 | 245 | 248 | 252 | ||||
| 68 | 103 | 104 | 159 | 215 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 216 | 232 | 236 | 245 | 248 | 252 | |||
| 20 | 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 255 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 256 | |||
| 68 | 103 | 104 | 159 | 232 | 236 | 245 | 248 | 252 | 260 | |||
| 68 | 103 | 104 | 159 | 228 | 232 | 236 | 245 | 248 | 252 | |||
| 68 | 76 | 89 | 103 | 104 | 159 | 210 | 213 | 232 | 236 | 245 | 260 | |
| 68 | 103 | 104 | 159 | 218 | 232 | 236 | 245 | 248 | 252 |
这些氨基酸位置编号是指那些分配给图1所示成熟解淀粉芽孢杆菌枯草杆菌蛋白酶序列的编号。然而，本发明不限于该特定枯草杆菌蛋白酶的突变，而是扩展到在一定位置处含有这样的氨基酸残基的前体蛋白酶，该残基“等同”于解淀粉芽孢杆菌枯草杆菌蛋白酶中特别确定的残基。在本发明的一个优选实施方案中，前体蛋白酶是迟缓芽孢杆菌枯草杆菌蛋白酶，并在迟缓芽孢杆菌中对应于以上所列位点的等同氨基酸残基位置处进行置换。
如果前体蛋白酶的残基(氨基酸)位置与解淀粉芽孢杆菌枯草杆菌蛋白酶中的特定残基或该残基的一部分同源(即，在一级或三级结构中位置相对应)或类似(即，具有相同或相似的化学结合、反应或相互作用的功能能力)，则它相当于解淀粉芽孢杆菌枯草杆菌蛋白酶的残基。
为了建立与一级结构的同源性，将前体蛋白酶的氨基酸序列直接与解淀粉芽孢杆菌枯草杆菌蛋白酶一级序列相比较，特别与公知在序列已知的枯草杆菌蛋白酶中不变的一组残基相比较。例如，在此图2显示解淀粉芽孢杆菌枯草杆菌蛋白酶与迟缓芽孢杆菌枯草杆菌蛋白酶之间的保守残基。对比保守残基，为维持序列对比进行必要的插入和缺失(即，避免保守残基由于任意缺失和插入而消除)之后，确定等同于解淀粉芽孢杆菌枯草杆菌蛋白酶一级序列中特定氨基酸的残基。保守残基的对比优选地应当保持100％的这些残基。然而，超过75％或低至50％的保守残基的对比也足以确定等同的残基。催化三联体Asp32/His64/Ser221的保守性应当保持。Siezen等人(1991)蛋白质工程(Protein Eng.)4(7):719-737显示了大量丝氨酸蛋白酶的序列对比。Siezen等人将此分组称为枯草杆菌酶(Subtilase)或枯草杆菌蛋白酶样丝氨酸蛋白酶。
例如，图3中，排列对比了解淀粉芽孢杆菌、枯草芽孢杆菌、地衣芽孢杆菌(carlsbergensis)和迟缓芽孢杆菌的枯草杆菌蛋白酶之氨基酸序列，以提供氨基酸序列之间最大量的同源性。这些序列的比较显示，在每种序列中含有许多保守残基。图2中确定了这些保守残基(BPN’与迟缓芽孢杆菌之间)。
因此，可用这些保守残基确定其它枯草杆菌蛋白酶，诸如迟缓芽孢杆菌枯草杆菌蛋白酶(1989年7月13日公布的PCT公开文本WO89/06279)、此处优选的蛋白酶前体酶或被称为PB92的枯草杆菌蛋白酶(EP0 328 299，它与优选的迟缓芽孢杆菌枯草杆菌蛋白酶高度同源)中与解淀粉芽孢杆菌枯草杆菌蛋白酶相应的等同氨基酸残基。在图3A和3B中将这些枯草杆菌蛋白酶中某些氨基酸序列与解淀粉芽孢杆菌枯草杆菌蛋白酶的序列相对比，产生保守残基的最大同源性。如所能看到的，与解淀粉芽孢杆菌枯草杆菌蛋白酶相比，迟缓芽孢杆菌序列中存在大量缺失。因此，例如，在其它枯草杆菌蛋白酶中与解淀粉芽孢杆菌枯草杆菌蛋白酶中的Val165等同的氨基酸对于迟缓芽孢杆菌和地衣芽孢杆菌而言是异亮氨酸。
也可以通过测定已经通过x-射线晶体分析法确定三级结构的前体蛋白酶在三级结构水平上的同源性，来确定“等同的残基”。等同的残基被定义为前体蛋白酶和解淀粉芽孢杆菌枯草杆菌蛋白酶特定氨基酸残基的两个或多个主链原子的原子坐标(N对N、CA对CA、C对C和O对O)经对比后在0.13nm优选地0.1nm之内的残基。在定向并定位最佳模型之后完成对比，得到所述蛋白酶与解淀粉芽孢杆菌枯草杆菌蛋白酶的非氢蛋白质原子的原子坐标的最大重叠。最佳模型是在可获得的最高分辨力时对于实验衍射数据可得到最低R因子的晶体模型。
与解淀粉芽孢杆菌枯草杆菌蛋白酶特定残基功能类似的等同残基被定义为前体蛋白酶的如下氨基酸，它们可采取一种构象，使得它们以一种确定的方式改变、修饰或贡献于蛋白质结构、底物结合或催化，且其归因于解淀粉芽孢杆菌枯草杆菌蛋白酶的特定残基。此外，它们是前体蛋白酶的那类残基(通过x-射线晶体分析法已获得其三级结构)，其占据了类似位点，使得尽管基于占据同源位点，特定残基的主链原子也许不能满足等同的标准，但该残基的至少两个侧链原子之原子坐标位于解淀粉芽孢杆菌枯草杆菌蛋白酶相应侧链原子的0.13nm内。在EPO公开文本0 251 446(相当于美国专利5,182,204，其公开内容在此引用作为参考)中阐述了解淀粉芽孢杆菌枯草杆菌蛋白酶的三维结构的坐标，并能如上所述用来在三级结构水平上确定等同的残基。
为了置换所确定的某些残基是保守残基，而其它的则不是。对于不保守的残基而言，一种或多种氨基酸的置换限于产生这样一种变体的置换，该变体具有一种与自然中所发现的氨基酸序列不对应的序列。对于保守残基而言，这些置换不能产生天然存在的序列。本发明的蛋白酶变体包括蛋白酶变体的成熟形式，以及这些蛋白酶变体的原-形式和前原-形式。前原-形式是优选的结构，因为其有利于蛋白酶变体的表达、分泌和成熟。
“原序列”是指与蛋白酶成熟形式的N端部分结合的氨基酸序列，当它被去除后，导致该蛋白酶“成熟”形式的出现。实质上发现许多蛋白水解酶为翻译酶原产物，在不存在翻译后加工的情况下以该方式表达。用于产生蛋白酶变体的一种优选原序列是解淀粉芽孢杆菌枯草杆菌蛋白酶的推定的原序列，尽管也可使用其它蛋白酶原序列。
“信号序列”或“前序列”是指与蛋白酶N端部分结合或与原蛋白酶N端部分结合的任何氨基酸序列，它们可能参与蛋白酶成熟形式或原形式的分泌。信号序列的这种定义是一种功能性定义，意为包括蛋白酶基因N端部分所编码的所有氨基酸序列，其在天然条件下参与蛋白酶分泌的实现。本发明利用这些序列实现此处所述蛋白酶变体的分泌。一种可能的信号序列包含枯草芽孢杆菌枯草杆菌蛋白酶信号序列的前7个氨基酸残基，这些残基与迟缓芽孢杆菌枯草杆菌蛋白酶信号序列(ATCC21536)的剩余部分相融合。
蛋白酶变体的“前原”形式由含有与蛋白酶氨基端有效连接的原序列的蛋白酶成熟形式和与原序列氨基端有效连接的“前”序列或“信号”序列组成。
“表达载体”是指一种DNA构建体，其含有与能实现该DNA在适当宿主中表达的适当控制序列有效连接的DNA序列。这类控制序列包括实现转录的启动子、控制这种转录的任意操纵子序列、编码适当mRNA核糖体结合位点的序列和控制转录与翻译终止的序列。载体可以是质粒、噬菌体颗粒，或者仅仅是有效的基因组插入片段。一旦转化到适当宿主中，载体可不依赖于宿主基因组复制并起作用，或者在某些情况中可能整合到基因组自身中。在本说明书中，“质粒”和“载体”有时交换使用，因为质粒是目前最常用的载体形式。然而，本发明意在包括表达载体的其它形式，它们具有等同的功能，并且是或者可成为本领域所公知的。
本发明中所用的“宿主细胞”通常是原核或真核宿主，它们优选地用美国专利RE34,606中公开的方法操作，以使其不能分泌酶促活性的内切蛋白酶。表达蛋白酶的一种优选宿主细胞是芽孢杆菌株BG2036，它缺乏具有有酶活性的中性蛋白酶和碱性蛋白酶(枯草杆菌蛋白酶)。美国专利5,264,366中详述了菌株BG2036的构建。表达蛋白酶的其它宿主细胞包括枯草芽孢杆菌I168(在美国专利RE34,606和美国专利5,264,366中也有描述，其公开内容在此引用作为参考)，以及任何合适的芽孢杆菌属的菌株，如地衣芽孢杆菌、迟缓芽孢杆菌等。
用以重组DNA技术构建的载体转化或转染宿主细胞。这些转化的宿主细胞能复制编码蛋白酶变体的载体或表达希望的蛋白酶变体。对于编码蛋白酶变体前-形式或前原-形式的载体而言，这些变体当表达时一般由宿主细胞向宿主细胞培养基中分泌。
当描述两个DNA区域之间的关系时，“有效连接”仅意味着它们在功能上彼此相关。例如，如果一种前序列作为信号序列参与蛋白质成熟形式的分泌，最可能的是参与信号序列的切割，则该前序列与肽有效连接。如果一种启动子控制一种编码序列的转录，则该启动子与该序列有效连接；如果一种核糖体结合位点位于一定位置使得翻译成为可能，则它与编码序列有效连接。
编码天然存在的前体蛋白酶的基因可依照本领域技术人员公知的通用方法获得。这些方法一般包括：合成含有编码目标蛋白酶区推定序列的标记探针、由表达该蛋白酶的生物制备基因组文库，以及通过与该探针杂交为了获得目标基因筛选该文库。然后对阳性杂交克隆作图并测定。
然后用克隆的蛋白酶转化宿主细胞，以表达该蛋白酶。然后将蛋白酶基因连接于高拷贝数质粒中。该质粒在宿主中复制，在此意义上它含有质粒复制所必需的众所周知的元件：与所述基因有效连接的启动子(如果它可被宿主识别，即转录，则可作为基因本身的同源启动子)、转录终止区和聚腺苷酸化区(在某些真核宿主细胞中，它对于由宿主从蛋白酶基因转录的mRNA的稳定性是必需的)，它是外源的或由蛋白酶基因的内源终止区所提供的，质粒中还期望含有选择性基因，如抗生素抗性基因，它使得质粒感染的宿主细胞通过在含有抗生素的培养基中生长以维持连续培养成为可能。高拷贝数质粒也含有宿主的复制起点，从而能在细胞质中不受染色体限制地产生大量质粒。然而，宿主基因组中整合多拷贝的蛋白酶基因也处于本发明之内。利用对同源重组特别敏感的原核和真核生物利于此过程。
该基因可能是天然迟缓芽孢杆菌基因。此外，也可以产生编码天然存在的或突变的前体蛋白酶的合成基因。这种方法中，测定前体蛋白酶的DNA和/或氨基酸序列。之后合成多个、重叠的合成单链DNA片段，杂交并连接后产生编码前体蛋白酶的合成DNA。在美国专利5,204,015的实施例3中描述了合成的基因构建体的一个实例，其公开内容在此引用作为参考。
一旦克隆了天然存在的或合成的前体蛋白酶基因，即进行大量修饰，以增强天然存在的前体蛋白酶合成之外的基因用途。这些修饰包括如美国专利RE34,606和EPO公布号0 251 446所公开的重组蛋白酶的产生和此处所述的蛋白酶变体的产生。
可用下列盒式诱变法促进本发明的蛋白酶变体的构建，尽管也可使用其它方法。首先，获得天然存在的编码蛋白酶的基因，并全部或部分测序。然后为了寻找希望在编码的酶中进行一个或多个氨基酸突变(缺失、插入或置换)的位点，扫描该序列。评价侧翼于该位点的序列是否存在估计限制酶切位点的存在，用于将基因的短片段替换为表达时编码不同突变体的寡核苷酸集合体。这些限制酶切位点优选地是蛋白酶基因内的唯一位点，以便于基因片段的置换。然而，可使用在蛋白酶基因中不过度丰余的任何适宜的限制酶切位点，只要限制酶切消化产生的基因片段能够在适当的序列中重新装配即可。如果在与所选位点适当距离内(10-15个核苷酸)的位置处不存在限制酶切位点，则可通过以在最终构建中不改变读框也不改变所编码氨基酸的方式置换基因中的核苷酸来产生这些位点。为了改变其序列以符合目标序列，基因突变依照公知的方法通过M13引物延伸来实现。根据遗传密码的丰余性、基因的限制酶图谱和大量不同的限制酶，常规进行定位适当侧翼区并估计所需改变以得到两种适宜的限制酶切位点序列的工作。注意到如果可获得适宜的侧翼限制酶切位点，则上述方法需要仅仅与不含该位点的侧翼区结合应用。
一旦克隆了天然存在的DNA或合成的DNA，即用同源的限制酶消化侧翼于待突变位点的限制酶切位点，并将多数末端互补的寡核苷酸盒连接于该基因中。该方法简化了诱变，因为能合成所有寡核苷酸使其具有相同的限制酶切位点，且合成的接头不是产生限制酶切位点所必需的。
在此使用时，蛋白水解活性被定义为每毫克活性酶的肽键水解率。测定蛋白水解活性有许多众所周知的方法(K.M.Kalisz，“微生物蛋白酶”《生物化学工程/生物技术进展》(Advances in BiochemicalEngineering/Biotechnology),A.Fiechter编，1988)。除了改进的蛋白水解活性之外，或者作为一种备择，本发明的变异蛋白酶可能具有其它改进的性质，如Km、kcat或kcat/Km比值和/或改进的底物特异性和/或改进的pH活性分布曲线。这些酶能量身定做般地用于例如预计在肽制剂中存在的特定底物，或适于水解过程如洗衣用途。
本发明的一个方面，目的在于获得一种与前体蛋白酶相比，在至少一种去污剂配方中和、或在至少一种洗涤条件下具有改变的，优选的改善的洗涤性能的蛋白酶变体。
洗涤条件多种多样，包括不同的去污剂配方、洗涤用水体积、洗涤用水温度和可接触蛋白酶变体的洗涤时间。例如，在不同领域中使用的去污剂配方在洗涤用水(wash water)中含有不同浓度的有关成分。例如，欧洲去污剂体系一般在洗涤用水中含有大约4500-5000ppm的去污剂成分，而日本去污剂体系一般在洗涤用水中含有大约667ppm的去污剂成分。在北美洲，尤其在美国，去污剂体系一般在洗涤用水中含有约975ppm的去污剂成分。
低去污剂浓度体系包括在洗涤用水中存在低于大约800ppm的去污剂成分的去污剂。日本去污剂一般被认为是低去污剂浓度体系，因为它们在洗涤用水中存在大约667ppm的去污剂成分。
中去污剂浓度包括在洗涤用水中存在大约800ppm到约2000ppm的去污剂成分的去污剂。北美去污剂一般被认为是中去污剂浓度体系，因为它们在洗涤用水中存在大约975ppm的去污剂成分。巴西的体系一般在洗涤用水中存在大约1500ppm的去污剂成分。
高去污剂浓度体系包括在洗涤用水中存在高于大约2000ppm的去污剂成分的去污剂。欧洲去污剂一般被认为是高去污剂浓度体系，因为它们在洗涤用水中存在大约4500-5000ppm的去污剂成分。
拉丁美洲去污剂一般是高泡沫磷酸盐助洗去污剂，在拉丁美洲使用的去污剂范围可能处于中及高去污剂浓度，因为在洗涤用水中它们有1500ppm到6000ppm的去污剂成分。如上所述，巴西的体系一般在洗涤用水中含有大约1500ppm的去污剂成分。然而，其它高泡沫磷酸盐助洗去污剂地区，不限于其它拉丁美洲国家，可能有在洗涤用水中存在可达大约6000ppm的去污剂成分的高去污剂浓度体系。
根据前述，显然典型洗涤溶液中去污剂组合物的浓度在全世界范围内各不相同，从低于大约800ppm的去污剂组合物(“低去污剂浓度地区”)，例如在日本为大约667ppm，到约800ppm-约2000ppm(“中去污剂浓度地区”)，例如在美国约为975ppm、在巴西约为1500ppm，到高于大约2000ppm(“高去污剂浓度地区”)，例如在欧洲为大约4500ppm-约5000ppm，在高泡沫助洗剂地区约为6000ppm。
典型洗涤溶液的浓度由经验确定。例如，在美国，一种典型的洗涤机可容纳大约64.4升体积的洗涤溶液。因此，为了获得在洗涤溶液中大约975ppm去污剂的浓度，必须向64.4升洗涤溶液中加入大约62.79克去污剂组合物。这种量是由消费者用去污剂所带量杯测量的加入洗涤用水中的典型量。
作为又一实例，不同地理区域使用不同洗涤温度。在日本的洗涤用水温度一般低于在欧洲使用的温度。
根据本发明的一个方面，包括了在至少一种洗涤条件下显示具有改善的洗涤性能的蛋白酶变体。
本发明的另一方面中，已确定在选自相应于解淀粉芽孢杆菌枯草杆菌蛋白酶之62、212、230、232、252和257残基位置的氨基酸置换对于酶之洗涤性能的改善十分重要。
这些置换优选地在迟缓芽孢杆菌(重组的或天然型)枯草杆菌蛋白酶中进行，尽管该置换也可在任何芽孢杆菌蛋白酶，优选地在芽孢杆菌属的枯草杆菌蛋白酶中进行。
根据用变异蛋白酶获得的筛选结果，解淀粉芽孢杆菌枯草杆菌蛋白酶中的显著突变对于这些酶的蛋白水解活性、性能和/或稳定性以及这些变异酶的清洁或洗涤能力至关重要。
本发明的许多蛋白酶变体可用于配制不同的去污剂组合物或个人护理制剂如洗发剂或洗涤剂。许多公知的化合物是在含有本发明之蛋白酶突变体的组合物中有用的适当表面活性剂。其包括非离子、阴离子、阳离子或两性离子去污剂，如Barry J.Anderson的US4,404,128和Jiri Flora等人的US4,261,868中所公开的。一种合适的去污剂制剂是美国专利5,204,015(前文引用作为参考)的实施例7中所述的制剂。本领域技术人员熟悉能用作清洁用组合物的不同制剂。除了典型的清洁用组合物之外，易于理解本发明的蛋白酶变体也可用于使用天然或野生型蛋白酶的任何目的。因此，例如，这些变体能用于固体肥皂或液体肥皂的用途、餐具清洁(dishcare)剂、隐形眼镜清洁液或产品、肽水解、废物处理、纺织品用途、用作蛋白质生产中的融合-切割酶，等等。本发明的变体可在去污剂组合物中具有增强的性能(与前体相比)。在此使用时，在去污剂中的增强性能定义为某些酶敏感性污迹如草或血液的清除加强，如在标准洗涤循环后通过通用评估所测定的。
本发明的蛋白酶能以大约0.01％到大约5％(优选地0.1％-0.5％)的重量百分比水平配制于已知的pH6.5-12.0的粉末及液体去污剂中。这些去污剂清洁用组合物也能包含其它的酶如已知的蛋白酶、淀粉酶、纤维素酶、脂肪酶或糖苷内切酶，以及助洗剂和稳定剂。
本发明的蛋白酶向常规清洁用组合物中的添加不产生任何特殊的应用限制。换句话说，适合于去污剂的任何温度和pH也适合于该组合物，只要pH位于上述范围之内，而温度低于所述蛋白酶的变性温度。另外，本发明的蛋白酶也能在不含去污剂的清洁用组合物中使用，单独或与助洗剂和稳定剂结合使用。
本发明也涉及含有本发明的蛋白酶变体的清洁用组合物。清洁用组合物另外也可含有常用于清洁用组合物的添加剂。其能选自但不限于：漂白剂、表面活性剂、助洗剂、酶和漂白催化剂。本领域的一名普通技术人员可容易地理解何种添加剂适合包含于组合物中。此处提供的列表绝不是完全的，而只是作为适当添加剂的实例。本领域的一名普通技术人员可容易地理解，应当只使用那些与该组合物中的酶及其它成分如表面活性剂相适合的添加剂。
制成后，清洁用组合物中存在的添加剂的量为大约0.01％-约99.9％，优选地约1％-约95％，更优选地约1％-约80％。
本发明的变异蛋白酶能包含于动物饲料中，如作为动物饲料添加剂部分，例如，如US5,612,055、US5,314,692和US5,147,642所述。
本发明的一个方面是含有本发明的变异蛋白酶的用于纺织品处理的组合物。如出版物如RD216,034、EP134,267、US4,533,359和EP344,259中所述，能用该组合物处理如丝绸或毛。
以下以实施例的方式描述，而不应理解为对权利要求书范围的限制。
此处引用的所有出版物和专利在此都完全引入作为参考。实施例1
用本领域众所周知的方法生产并纯化大量蛋白酶变体。所有突变都在迟缓芽孢杆菌GG36枯草杆菌蛋白酶中进行。变体示于表4。
表4
| N76D | S103A | V104I | M222S | |||||||||
| N76D | A98E | S103A | V104I | |||||||||
| N76D | S78T | S103A | V104I | |||||||||
| N76D | S103A | V104I | I107V | |||||||||
| V4E | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | I246V | |||||||||
| N76D | N77D | S103A | V104I | |||||||||
| N76D | S103A | V104I | N183D | N218I | ||||||||
| A16T | N76D | S103A | V104I | N248D | ||||||||
| A1E | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | N261D | |||||||||
| N76D | S103A | V104I | S160T | |||||||||
| N76D | S103A | V104I | S216C | |||||||||
| H17Q | N76D | S103A | V104I | |||||||||
| S37T | N76D | S103A | V104I | |||||||||
| N76D | N77D | S103A | V104I | A174V |
表4(续)
| T38S | N76D | S103A | V104I | |||||||||
| T38S | N76D | S103A | V104I | K237Q | ||||||||
| I8V | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | N183D | |||||||||
| R19L | N76D | S103A | V104I | |||||||||
| A13V | N76D | S103A | V104I | |||||||||
| R19C | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | N184D | |||||||||
| N76D | S103A | V104I | N252D | |||||||||
| N76D | S103A | V104I | S259C | |||||||||
| N76D | S103A | V104I | K251T | |||||||||
| N76D | P86S | S103A | V104I | |||||||||
| I72V | N76D | S103A | V104I | N185D | ||||||||
| N76D | S103A | V104I | K237E | T274A | ||||||||
| N76D | S103A | V104I | S160L | |||||||||
| N76D | S103A | V104I | A228V | |||||||||
| P55S | N76D | S103A | V104I | S240T | ||||||||
| N76D | S103A | V104I | A254T |
表4(续)
| N76D | S103A | I104N | N204T | |||||||||
| N76D | S103A | V104I | N204D | |||||||||
| N43S | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | G159D | |||||||||
| R10H | N76D | S103A | V104I | V177A | ||||||||
| T58S | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | A270V | |||||||||
| N76D | S103A | V104I | N185D | |||||||||
| K27N | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | L262M | |||||||||
| N76D | S78P | S103A | V104I | |||||||||
| S24P | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | S166G | Q236R | K251R | |||||||
| H17L | N76D | S103A | V104I | K237E | ||||||||
| N76D | S103A | V104I | S130L | |||||||||
| N76D | S103A | V104I | Q109R | |||||||||
| N76D | S99R | S103A | V104I | N204T | ||||||||
| N76D | S103A | V104I | D181N |
表4(续)
| Q12R | N76D | S103A | V104I | |||||||||
| N76D | S103A | V104I | S212P | E271V | ||||||||
| N76D | S103A | V104I | N252K | N261Y | ||||||||
| N76D | S103A | V104I | S242T | |||||||||
| N76D | S103A | V104I | E271Q | |||||||||
| Q12R | N76D | S103A | V104I | S242T | ||||||||
| N43S | N76D | S103A | V104I | N116K | N183I | |||||||
| N76D | S103A | V104I | G258R | |||||||||
| N76D | S103A | V104I | E271G | |||||||||
| G61R | N76D | S103A | V104I | |||||||||
| T38S | N76D | S103A | V104I | Q182R | Y263H | |||||||
| N76D | S103A | V104I | Q182R | A272S | ||||||||
| N76D | S103A | V104I | Q109R | I246V | ||||||||
| N76D | S87G | S103A | V104I | Q206R | H249Q | S265G | ||||||
| N76D | S103A | V104I | Q137R | N238Y | E271V | |||||||
| S103A | V104I | A228T | ||||||||||
| N76D | S103A | V104I | Q182R | I198V | ||||||||
| L21M | N76D | S103A | V104I | Q182R |
表4(续)
| N76D | S103A | V104I | M119I | Q137R | ||||||||
| N76D | S103A | V104I | Q137R | N248S | ||||||||
| A13T | N76D | S103A | V104I | Q206R | ||||||||
| N76D | S103A | V104I | Q206R | |||||||||
| N76D | S103A | V104I | S212P | G258R | ||||||||
| T58S | N76D | S103A | V104I | E271G | ||||||||
| N76D | S103A | V104I | Q206E | N261D | ||||||||
| V4E | N76D | S103A | V104I | Q206E | ||||||||
| N76D | N77D | S103A | V104I | Q206E | ||||||||
| N76D | S103A | V104I | A158E | |||||||||
| N76D | S103A | V104I | Q206E | |||||||||
| V4E | N76D | S103A | V104I | G159D | L217E | K251Q | ||||||
| V4E | N76D | S103A | V104I | G159D | L217E | N252D | ||||||
| N76D | N77D | S103A | V104I | A133T | N185D | K251T | ||||||
| N76D | S103A | V104I | G159D | Q206E | V244A | |||||||
| V4E | N76D | S103A | V104I | S188E | ||||||||
| V4E | N76D | S103A | V104I | A158E | ||||||||
| N76D | N77D | S103A | V104I | N185D |
表4(续)
| N76D | S103A | V104I | Q206E | K251T | ||||||||
| A48T | N76D | S103A | V104I | L111M | G159D | |||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | |||||||
| L42V | N76D | S103A | V104I | G159D | ||||||||
| Q12H | N62H | N76D | S103A | V104I | G159D | |||||||
| L42I | N76D | S103A | V104I | G159D | ||||||||
| N76D | S103A | V104I | G146S | G159D | ||||||||
| N76D | S103A | V104I | G159D | N238S | ||||||||
| N76D | S103A | V104I | G159D | T224A | ||||||||
| N76D | S103A | V104I | S212P | V268F | E271V | |||||||
| N76D | E89A | S103A | V104I | |||||||||
| N76D | S87R | S103A | V104I | S212P | E271V | |||||||
| N76D | S103A | V104I | S212P | Q245L | E271V | |||||||
| N76D | S103A | V104I | T134S | S141N | S212P | E271V | ||||||
| N76D | S103A | V104I | S212P | Q236L | N243S | E271V | ||||||
| N76D | S103A | V104I | Q109R | Q245R | ||||||||
| N76D | S103A | V104I | Q109R | P210L | ||||||||
| G20V | N62S | N76D | S103A | V104I |
表4(续)
| V68A | N76D | S103A | V104I | Q236H | ||||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | E271V | ||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | ||||||
| V68A | N76D | S103A | V104I | G159D | L217I | Q236H | E271V | |||||
| H17Q | V68A | N76D | S103A | V104I | ||||||||
| V68A | N76D | S103A | V104I | |||||||||
| V68A | N76D | S103A | V104I | G159D | Q236R | |||||||
| V68A | L75R | N76D | S103A | V104I | G159D | Q236H | ||||||
| V68A | N76D | N76D | S103A | A114V | V121I | G159D | Q236H | Q245R | ||||
| Q12R | V68A | N76D | S103A | V104I | G159D | Q236H | ||||||
| V68A | N76D | S103A | V104I | G159D | Y209S | Q236H | T253K | |||||
| V68A | N76D | S103A | V104I | N117K | G159D | N184S | Q236H | |||||
| V68A | N76D | S103A | V104I | G159D | Q236H | N243I | ||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245L | ||||||
| V68A | N76D | S103A | V104I | A142V | G159D | |||||||
| V68A | N76D | S103A | V104I | N123S | G159D | Q236H | H249Y | |||||
| V68A | N76D | S103A | V104I | G159D | Q236H | H249Q | ||||||
| N76D | S103A | V104I | M222S | Q245R |
表4(续)
| N76D | S103A | V104I | Q12R | M222S | H249R | |||||||
| N76D | S103A | V104I | N173R | M222S | ||||||||
| N76D | S103A | V104I | M222S | Y263F | ||||||||
| L21M | N76D | S103A | V104I | M222S | K237R | Y263F | ||||||
| N76D | S103A | V104I | Q109R | M222S | ||||||||
| N76D | S103A | V104I | Q109R | M222S | E271D | |||||||
| G61R | N76D | S103A | V104I | M222S | ||||||||
| N76D | S103A | V104I | Q137R | M222S | ||||||||
| N76D | S103A | V104I | Q109R | M222S | N248S | |||||||
| N76D | S103A | V104I | M222S | H249R | ||||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | N261D | |||||
| V68A | N76D | S103A | V104I | S141N | G159D | Q236H | Q245R | T255S | ||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | R247H | |||||
| V68A | N76D | S103A | V104I | G159D | A174V | N204D | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | N204D | Q236H | Q245R | |||||
| V68A | N76D | S103A | V104I | A133V | G159D | N218D | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | N76D | S103A | V104I | G159D | A194I | V203A | Q236H | Q245R |
表4(续)
| Q12R | N76D | S103A | V104I | M222S | Q245R | |||||||
| N76D | S103A | V104I | A232V | Q245R | ||||||||
| S24T | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | |||||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | |||
| Q12R | N76D | S103A | I104T | M222S | V244I | Q245R | ||||||
| Q12R | N76D | S103A | M222S | P210T | Q245R | |||||||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | ||||||
| T22K | V68A | N76D | S103A | V104I | ||||||||
| V68A | N76D | S103A | N184D | |||||||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||||
| V68A | S103A | V104I | N140D | G159D | A232V | Q236H | Q245R | N252K | ||||
| N43S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | ||||
| N43K | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | ||||
| V68A | S87G | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | R275S | |||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | N248S | L262M |
表4(续)
| Q12R | N76D | S103A | I104T | S130T | A215V | M222S | Q245R | |||||
| Q12R | N76D | S103A | I104T | S130T | M222S | V227A | Q245R | L262S | ||||
| Q12R | N76D | S103A | I104T | S130T | A215T | M222S | Q245R | |||||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | N261D | |||||
| N76D | S103A | I104T | S130T | M222S | Q245R | |||||||
| Q12R | N76D | S103A | I104T | S130T | N218D | M222S | Q245R | L262S | N269D | |||
| Q12R | S57P | N76D | S103A | I104T | S130T | M222S | Q245R | K251Q | ||||
| Q12R | N76D | S103A | I104T | S130T | R170S | N185D | M222S | N243D | Q245R | |||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | V268A | |||||
| Q12R | N76D | S103A | I104T | S130T | M222S | P210S | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | |||||
| V68A | S103A | V104I | N116D | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | |||||
| R10C | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | V203E | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | K237E | Q245R | |||||
| V68A | N76D | I79N | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | N183D | A232V | Q236H | Q245R |
表4(续)
| V68A | S103A | V104I | G159D | A174V | Q206L | A232V | Q236H | Q245R | ||||
| V68A | S103A | V104I | G159D | S188C | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A230T | A232V | Q236H | Q245R | |||||
| V68A | A98T | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A215T | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248S | |||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | |||||
| V68A | N76D | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | ||||
| N76D | S103A | V104I | A232V | Q236H | Q245R | L257V | ||||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | R275H | ||||
| N76D | S103A | V104I | L257V | R275H | ||||||||
| V68A | S103A | V104I | G159D | T224A | A232V | Q236H | Q245R | L257V | ||||
| N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | |||||
| V68A | N76D | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | G211R | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | G159D | G211V | A232V | Q236H | Q245R | ||||
| Q12R | V68A | N76D | S103A | V104I | G159D | Y214L | A232V | Q236H | Q245R |
表4(续)
| V68A | N76D | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | ||||
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| G20R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S259G | |||
| V68A | S87R | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | T260V | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261G | ||||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261W | ||||
| N76D | S103A | V104I | A232V | Q236H | S242P | Q245R | ||||||
| V68A | N76D | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | ||||
| Q12R | A48V | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | |||
| N76D | S103A | V104I | A232V | Q236H | Q245R | |||||||
| N76D | S103A | V104I | G159D | Y192F | A232V | Q236H | Q245R | |||||
| N76D | S103A | V104I | V147I | G159D | A232V | Q236H | Q245R | N248S | K251R | |||
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | A272S | |||
| V68A | N76D | S103A | V104I | G159D | N183K | Q206L | A232V | Q236H | Q245R | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S256R | ||||
| V68A | N76D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | ||||
| K27R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | ||||
| V68A | N76D | S103A | V104I | N116T | G159D | R170S | N185S | A232V | Q236H | Q245R |
表4(续)
| G61E | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212P | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | S99N | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | ||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| G20R | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | Y209F | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261D | |||
| V68A | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210T | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | P210S | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | N185D | P210L | A232V | Q236H | Q245R | N248D | N252K | ||
| V68A | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212A | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K |
表4(续)
| V68A | S103A | V104I | G159D | S212E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | T213S | A232V | Q236H | Q245R | N248D | N252K | ||||
| V68A | A103V | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | T213G | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A215V | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S216T | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S216V | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | S216C | A232V | Q236H | Q245R | N248D | N252K | |||
| G20A | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | N173D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | K251V | N252K | |||
| V68A | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252L | ||||
| P55S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F |
表4(续)
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T255V | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256N | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256E | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T260R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | L257R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | G258D | |||
| I8V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N269D | ||
| V68A | S103A | V104I | N116S | G159D | A232V | Q236H | Q245R | N248D | N252K | T260E | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261D | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | |||||
| S103A | V104I | G159D | A232S | Q236H | Q245R | N248D | N252K | |||||
| V68A | S103A | V104I | G159D | A232V | Q236R | Q245R | N248D | N252K | ||||
| N18S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245V | N248D | N252K | ||||
| V68A | N76D | S101T | S103A | V104I | G159D | T213R | N218S | A232V | Q236H | Q245R | T260A |
表4(续)
| V68A | S103A | V104I | G159D | A228V | A232V | Q236H | Q245R | N248D | N252K | |||
| T33S | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| V68A | N76D | E89D | S103A | V104I | G159D | P210L | T213R | A232V | Q236H | Q245R | T260A | |
| G61E | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| S103A | V104I | G159D | V205I | P210I | A232V | Q236H | Q245R | |||||
| G61E | V68A | S103A | V104I | S130A | G159D | A232V | Q236H | Q245R | N248D | N252K | ||
| G61E | V68A | S103A | V104I | A133S | Q137R | G159D | A232V | Q236H | Q245R | N248D | N252K | |
| G61E | S103A | V104I | A133V | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248G | N252K | ||||
| V68A | S103A | V104I | G159D | N218S | A232V | Q236H | Q245R | N248D | N252K | |||
| G61E | V68A | S103A | V104I | G159D | S160V | A232V | Q236H | Q245R | N248D | N252K | ||
| S3L | G61E | V68A | N76D | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | ||
| G61E | V68A | S103A | V104I | G159D | S167F | A232V | Q236H | Q245R | N248D | N252K | ||
| G97E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| A98D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S99E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S101E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K |
表4(续)
| G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | S106E | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | Q109E | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | ||||
| S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| N62D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | S166D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | L217E | A232V | Q236H | Q245R | N248D | N252K | ||||
| G20R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| S103A | V104I | G159D | Q206R | L217E | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | |||
| S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| K27N | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| T38G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| T38A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A |
表4(续)
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | E271G | ||
| V68A | N76D | S103A | V104I | G159D | Y209W | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | P210I | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | V205I | T213R | A232V | Q236H | Q245R | T260A | ||
| V68A | N76D | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | T260A | |||
| V68A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | ||||
| N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | ||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | A230V | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | L126F | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | |||||
| S103A | V104I | G159D | A230V | Q236H | Q245R | |||||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | T260A | |||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | |||||||
| V68A | S103A | V104I | G159D | A174V | A232V | Q236H | Q245R | L257V | ||||
| V68A | S103A | V104I | G159D | A194S | A232V | Q236H | Q245R | L257V |
表4(续)
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | L257V | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | ||||||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | N261W | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | N261W | ||||
| S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | |||||
| S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | L257V | |||||
| V68A | N76D | S103A | V104I | G159D | P210L | T213R | A232V | Q236H | Q245R | T260A | ||
| Q12R | S103A | V104I | G159D | Y209W | T213R | A232V | Q236H | Q245R | T260A | |||
| S103A | V104I | Y209W | A232V | Q236H | Q245R | L257V | ||||||
| S103A | V104I | G159D | V205I | P210I | T213R | A232V | Q236H | Q245R | T260A | |||
| S103A | V104I | G159D | V205I | Y209W | A232V | Q236H | Q245R | T260A | ||||
| V68A | S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | |||
| S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | L257V | |||
| S103A | V104I | G159D | V205I | Y209W | A232V | Q236H | Q245R | L257V | ||||
| V68A | S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | T260A | ||
| S103A | V104I | G159D | V205I | Y209W | P210I | A232V | Q236H | Q245R | ||||
| S103A | V104I | G159D | Y209W | P210I | A232V | Q236H | Q245R |
表4(续)
| S103A | V104I | G159D | V205I | P210I | A232V | Q236H | Q245R | |||||
| V68A | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | |||||
| A48V | S103A | V104I | G159D | A230V | Q236H | Q245R | ||||||
| A48V | V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | ||||
| A48V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A48V | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | N261W | |||
| G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | |||
| Q12R | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| G102A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | |||
| S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184S | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | N184G | A232V | Q236H | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | V244T | Q245R | N248D | N252K | ||||
| S103A | V104I | G159D | A232V | Q236H | V244A | Q245R | N248D | N252K | ||||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | S256R | ||
| Q12R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K |
表4(续)
| S101G | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | Q206E | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | T213Q | A232V | Q236H | Q245R | N248D | N252K | |||
| A98L | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | N248D | N252K | |||
| N62D | S103A | V104I | Q109R | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | G159D | S212G | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S101G | S103A | V104I | G159D | S212G | T213R | A232V | Q236H | Q245R | N248D | N252K | |
| S103A | V104I | G159D | A232V | Q245R | N248D | N252K | ||||||
| S103A | V104I | G159D | A230V | Q245R | ||||||||
| N62D | S103A | V104I | S130G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | S128G | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S101G | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| N62D | S103A | V104I | S128G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K |
表4(续)
实施例2
| N62D | S103A | V104I | S128L | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T260A | |||
| S101G | S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| A98V | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S99G | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | N248D | N252K | |||
| S101G | S103A | V104I | G159D | A230V | Q236H | Q245R | ||||||
| S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | |||
| N76D | S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | ||
| S101G | S103A | V104I | G159D | A230V | A232V | Q236H | Q245R | N248D | N252K | |||
| N62D | S103A | V104I | G159D | N185D | Q206E | T213R | A232V | Q236H | Q245R | N248D | N252K | E271Q |
用美国系列号60/068,796“一种测定优选的酶和/或优选的去污剂组合物的改进方法”中所述的微观(microswatch)测定法检测如实施例1所述制备的大量蛋白酶变体在两种类型的去污剂及洗涤条件中的表现。
表5中列出检测的变体以及在两种不同去污剂中测试的结果。A栏；0.67g/l过滤的Ariel Ultra(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有3格令混合Ca2+/Mg2+硬度的溶液中，20℃下每孔使用0.3ppm的酶。B栏；3.38g/l过滤的Ariel Futur(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有15格令混合Ca2+/Mg2+硬度的溶液中，40℃下每孔使用0.3ppm的酶。
表5
表5(续)
表5(续)
表5(续)
表5(续)
表5(续)
表5(续)
表5(续)
表5(续)
表5(续)
实施例3
| A | B | |||||||||||
| N76D | S103A | V104I | 1 | 1 | ||||||||
| S103A | V104I | A228T | 0.56 | 1.11 | ||||||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | 1.41 | 1.85 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.77 | 1.20 | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.26 | 1.67 | ||||
| V68A | S103A | V104I | N140D | G159D | A232V | Q236H | Q245R | N252K | 2.96 | 1.42 | ||
| N43S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | 1.91 | 1.80 | ||
| N43K | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.05 | 1.78 | |||
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252K | 2.00 | 1.34 | ||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | 2.38 | 1.67 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | 2.83 | 0.53 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | K237E | Q245R | 2.87 | 0.20 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N252S | 2.56 | 1.41 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | R275H | 3.97 | 0.47 | ||
| V68A | S103A | V104I | G159D | T224A | A232V | Q236H | Q245R | L257V | 3.35 | 1.28 | ||
| G61E | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.77 | 0.09 |
| N43D | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.50 | 0.47 | |
| V68A | S103A | V104I | G159D | S212P | A232V | Q236H | Q245R | N248D | N252K | 2.81 | 1.46 | |
| N76D | A98E | S103A | V104I | 1.56 | 0.28 | |||||||
| V4E | N76D | S103A | V104I | 1.22 | 0.33 | |||||||
| N76D | N77D | S103A | V104I | 1.13 | 0.36 | |||||||
| A16T | N76D | S103A | V104I | N248D | 1.22 | 0.43 | ||||||
| A1E | N76D | S103A | V104I | 1.12 | 0.32 | |||||||
| N76D | S103A | V104I | N261D | 1.54 | 0.33 | |||||||
| N76D | S103A | V104I | S216C | 1.04 | 0.13 | |||||||
| N76D | N77D | S103A | V104I | A174V | 1.09 | 0.35 | ||||||
| T38S | N76D | S103A | V104I | K237Q | 1.11 | 0.55 | ||||||
| N76D | S103A | V104I | N183D | 1.50 | 0.25 | |||||||
| R19L | N76D | S103A | V104I | 1.11 | 0.48 | |||||||
| R19C | N76D | S103A | V104I | 1.05 | 0.19 | |||||||
| N76D | S103A | V104I | N184D | 1.32 | 0.29 | |||||||
| N76D | S103A | V104I | N252D | 1.19 | 0.53 | |||||||
| N76D | S103A | V104I | S259C | 0.92 | 0.12 | |||||||
| N76D | S103A | V104I | K251T | 1.31 | 0.43 |
| N76D | P86S | S103A | V104I | 1.00 | 0.98 | |||||||
| I72V | N76D | S103A | V104I | N185D | 1.70 | 0.37 | ||||||
| N76D | S103A | V104I | K237E | T274A | 1.12 | 0.16 | ||||||
| N76D | S103A | V104I | A228V | 1.13 | 0.99 | |||||||
| N76D | S103A | V104I | G159D | 1.88 | 0.23 | |||||||
| H17L | N76D | S103A | V104I | K237E | 1.29 | 0.28 | ||||||
| N76D | S103A | V104I | S130L | 0.52 | 0.71 | |||||||
| N76D | S103A | V104I | Q109R | 0.23 | 1.26 | |||||||
| N76D | S99R | S103A | V104I | N204T | 0.21 | 0.87 | ||||||
| N76D | S103A | V104I | D181N | 0.24 | 1.07 | |||||||
| Q12R | N76D | S103A | V104I | 0.61 | 1.31 | |||||||
| N76D | S103A | V104I | S212P | E271V | 0.69 | 1.35 | ||||||
| N76D | S103A | V104I | N252K | N261Y | 0.37 | 1.02 | ||||||
| N76D | S103A | V104I | S242T | 0.98 | 0.92 | |||||||
| N76D | S103A | V104I | E271Q | 0.63 | 1.25 | |||||||
| Q12R | N76D | S103A | V104I | S242T | 0.49 | 1.32 | ||||||
| N43S | N76D | S103A | V104I | N116K | N183I | 0.39 | 1.10 | |||||
| N76D | S103A | V104I | G258R | 0.34 | 1.17 |
| N76D | S103A | V104I | E271G | 0.57 | 1.25 | |||||||
| N76D | S103A | V104I | Q182R | I198V | 0.22 | 0.95 | ||||||
| L21M | N76D | S103A | V104I | Q182R | 0.24 | 0.98 | ||||||
| N76D | S103A | V104I | M119I | Q137R | 0.13 | 0.91 | ||||||
| N76D | S103A | V104I | Q137R | N248S | 0.16 | 1.02 | ||||||
| A13T | N76D | S103A | V104I | Q206R | 0.31 | 1.01 | ||||||
| N76D | S103A | V104I | Q206R | 0.33 | 1.02 | |||||||
| N76D | S103A | V104I | S212P | G258R | 0.38 | 1.06 | ||||||
| T58S | N76D | S103A | V104I | E271G | 0.84 | 1.26 | ||||||
| N76D | S103A | V104I | Q206E | N261D | 1.97 | 0.04 | ||||||
| V4E | N76D | S103A | V104I | Q206E | 1.51 | 0.05 | ||||||
| N76D | N77D | S103A | V104I | Q206E | 1.40 | 0.04 | ||||||
| N76D | S103A | V104I | A158E | 1.95 | 0.16 | |||||||
| N76D | S103A | V104I | Q206E | 2.41 | 0.88 | |||||||
| N76D | N77D | S103A | V104I | A133T | N185D | K251T | 1.34 | 0.03 | ||||
| N76D | S103A | V104I | Q206E | N261D | 1.78 | 0.04 | ||||||
| N76D | S103A | V104I | G159D | Q206E | V244A | 2.16 | 0.04 | |||||
| V4E | N76D | S103A | V104I | S188E | 1.91 | 0.04 |
| V4E | N76D | S103A | V104I | A158E | 2.06 | 0.04 | ||||||
| N76D | S103A | V104I | Q206E | K251T | 1.73 | 0.06 | ||||||
| A48T | N76D | S103A | V104I | L11M | G159D | 2.04 | 0.16 | |||||
| V68A | N76D | S103A | V104I | G159D | Q236H | 3.20 | 0.09 | |||||
| L42V | N76D | S103A | V104I | G159D | 1.83 | 0.17 | ||||||
| Q12H | N62H | N76D | S103A | V104I | G159D | 1.42 | 0.14 | |||||
| L42I | N76D | S103A | V104I | G159D | 1.86 | 0.18 | ||||||
| N76D | S103A | V104I | G146S | G159D | 1.87 | 0.19 | ||||||
| N76D | S103A | V104I | G159D | N238S | 1.90 | 0.15 | ||||||
| N76D | S103A | V104I | G159D | T224A | 1.61 | 0.07 | ||||||
| N76D | S103A | V104I | S212P | V268F | E271V | 0.44 | 1.42 | |||||
| N76D | S87R | S103A | V104I | S212P | E271V | 0.39 | 2.03 | |||||
| N76D | S103A | V104I | S212P | Q245L | E271V | 0.62 | 1.79 | |||||
| N76D | S103A | V104I | Q109R | Q245R | 0.11 | 1.78 | ||||||
| N76D | S103A | V104I | Q109R | P210L | 0.12 | 1.21 | ||||||
| G20V | N62S | N76D | S103A | V104I | 1.63 | 0.78 | ||||||
| V68A | N76D | S103A | V104I | Q236H | 2.37 | 0.44 | ||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | E271V | 2.97 | 0.45 |
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | 3.00 | 0.61 | ||||
| V68A | N76D | S103A | V104I | G159D | L217I | Q236H | E271V | 2.71 | 0.12 | |||
| H17Q | V68A | N76D | S103A | V104I | 2.46 | 0.38 | ||||||
| V68A | N76D | S103A | V104I | 2.46 | 0.61 | |||||||
| V68A | N76D | S103A | V104I | G159D | Q236R | 3.31 | 0.11 | |||||
| V68A | L75R | N76D | S103A | V104I | G159D | Q236H | 3.06 | 0.14 | ||||
| V68A | N76D | S103A | V104I | A114V | V121I | G159D | Q236H | Q245R | 3.11 | 0.40 | ||
| Q12R | V68A | N76D | S103A | V104I | G159D | Q236H | 3.12 | 0.34 | ||||
| V68A | N76D | S103A | V104I | G159D | Y209S | Q236H | T253K | 3.18 | 0.03 | |||
| V68A | N76D | S103A | V104I | N117K | G159D | N184S | Q236H | 2.78 | 0.06 | |||
| V68A | N76D | S103A | V104I | Q236H | 2.49 | 0.57 | ||||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245L | 3.37 | 0.03 | ||||
| V68A | N76D | S103A | V104I | N123S | G159D | Q236H | H249Y | 3.11 | 0.03 | |||
| V68A | N76D | S103A | V104I | G159D | Q236H | H249Q | 3.15 | 0.04 | ||||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | N261D | 3.31 | 0.03 | |||
| V68A | N76D | S103A | V104I | S141N | G159D | Q236H | Q245R | T255S | 3.26 | 0.62 | ||
| V68A | N76D | S103A | V104I | G159D | Q236H | Q245R | R247H | 2.78 | 0.03 | |||
| V68A | N76D | S103A | V104I | G159D | A174V | N204D | Q236H | Q245R | 3.28 | 0.02 |
| V68A | N76D | S103A | V104I | G159D | N204D | Q236H | Q245R | 3.34 | 0.02 | |||
| V68A | N76D | S103A | V104I | A133V | G159D | N218D | Q236H | Q245R | 3.28 | 0.03 | ||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.91 | 0.58 | |||
| V68A | N76D | S103A | V104I | G159D | A194I | V203A | Q236H | Q245R | 2.86 | 0.13 | ||
| V68A | N76D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | T260A | 1.30 | 1.73 | |
| T22K | V68A | N76D | S103A | V104I | 1.83 | 1.13 | ||||||
| V68A | N76D | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | 1.28 | 1.54 | ||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | 3.72 | 0.8 | ||
| N76D | S103A | V104I | A232V | Q236H | Q245R | L257V | 0.6 | 1.5 | ||||
| N76D | S103A | V104I | L257V | R275H | 1.91 | 0.15 | ||||||
| N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | L257V | 1.92 | 1.09 | |||
| V68A | N76D | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | 3.57 | 0.99 | ||
| V68A | N76D | S103A | V104I | G159D | G211R | A232V | Q236H | Q245R | 1.74 | 1.76 | ||
| V68A | N76D | S103A | V104I | G159D | G211V | A232V | Q236H | Q245R | 3.15 | 1.06 | ||
| Q12R | V68A | N76D | S103A | V104I | G159D | Y214L | A232V | Q236H | Q245R | 2.33 | 1.92 | |
| V68A | N76D | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | 1.67 | 1.45 | ||
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.16 | 1.72 | ||
| G20R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S259G | 2.77 | 1.59 |
| V68A | N76D | S87R | S103A | V104I | G159D | A232V | Q236H | Q245R | T260V | 2.62 | 1.49 | |
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261G | 2.92 | 0.68 | ||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N261W | 2.17 | 1.37 | ||
| N76D | S103A | V104I | A232V | Q236H | S242P | Q245R | 0.48 | 1.2 | ||||
| V68A | N76D | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | 2.92 | 0.76 | ||
| Q12R | A48V | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.09 | 1.86 | |
| N76D | S103A | V104I | A232V | Q236H | Q245R | 0.51 | 1.44 | |||||
| N76D | S103A | V104I | G159D | Y192F | A232V | Q236H | Q245R | 1.60 | 1.14 | |||
| N76D | S103A | V104I | V147I | G159D | A232V | Q236H | Q245R | N248S | K251R | 1.35 | 1.29 | |
| Q12R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | A272S | 1.92 | 1.81 | |
| V68A | N76D | S103A | V104I | G159D | N183K | Q206L | A232V | Q236H | Q245R | 1.17 | 1.53 | |
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | S256R | 2.01 | 1.72 | ||
| V68A | N76D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | 2.09 | 1.62 | ||
| K27R | V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | 3.00 | 1.08 | ||
| V68A | N76D | S103A | V104I | N116T | G159D | R170S | N185S | A232V | Q236H | Q245R | ND | ND |
| N76D | S103A | V104I | M222S | Q245R | 1.01 | 1.23 | ||||||
| Q12R | N76D | S103A | V104I | M222S | H249R | 0.57 | 1.65 | |||||
| N76D | S103A | V104I | N173R | M222S | 0.86 | 0.46 |
| N76D | S103A | V104I | M222S | Y263F | 1.24 | 0.77 | ||||||
| L21M | N76D | S103A | V104I | M222S | K237R | Y263F | 1.18 | 0.76 | ||||
| N76D | S103A | V104I | Q109R | M222S | 0.52 | 1.16 | ||||||
| N76D | S103A | V104I | Q109R | M222S | E271D | 0.56 | 1.12 | |||||
| G61R | N76D | S103A | V104I | M222S | 0.43 | 0.96 | ||||||
| N76D | S103A | V104I | Q137R | M222S | 0.42 | 1.25 | ||||||
| N76D | S103A | V104I | M222S | H249R | 1.15 | 1.01 | ||||||
| Q12R | N76D | S103A | V104I | M222S | Q245R | 0.53 | 1.46 | |||||
| N76D | S103A | V104I | A232V | Q245R | 0.69 | 1.56 | ||||||
| Q12R | N76D | S103A | I104T | M222S | V244I | Q245R | 0.66 | 1.74 | ||||
| Q12R | N76D | S103A | V104I | M222S | P210T | Q245R | 0.52 | 1.56 | ||||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | 0.70 | 1.61 | ||||
| Q12R | N76D | S103A | I104T | S130T | A215V | M222S | Q245R | 0.79 | 1.85 | |||
| Q12R | N76D | S103A | I104T | S130T | M222S | V227A | Q245R | L262S | 0.78 | 1.56 | ||
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | N261D | 1.25 | 1.30 | |||
| N76D | S103A | I104T | S130T | M222S | Q245R | 1.29 | 1.30 | |||||
| Q12R | S57P | N76D | S103A | I104T | S130T | M222S | Q245R | K251Q | 1.44 | 0.16 | ||
| Q12R | N76D | S103A | I104T | S130T | R170S | N185D | M222S | N243D | Q245R | 2.01 | 0.04 |
| Q12R | N76D | S103A | I104T | S130T | M222S | Q245R | V268A | 0.77 | 1.60 | |||
| Q12R | N76D | S103A | I104T | S130T | M222S | P210S | Q245R | 0.73 | 1.66 | |||
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | 2.09 | 0.86 |
表6中列出检测的来自实施例1中的变体以及在四种不同去污剂中测试的结果。进行如实施例2中的相同操作考察了变体蛋白酶在下列去污剂中的洗涤性能。A栏；0.67g/l过滤的Ariel Ultra(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有3格令混合Ca2+/Mg2+硬度的溶液中，20℃下每孔使用0.3ppm的酶。B栏；3.38g/l过滤的Ariel Futur(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有15格令混合Ca2+/Mg2+硬度的溶液中，40℃下每孔使用0.3ppm的酶。C栏，3.5g/LHSP1去污剂(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有8格令混合Ca2+/Mg2+硬度的溶液中，20℃下每孔使用0.3ppm的酶D栏，1.5ml/l Tide KT去污剂(Procter&Gamble,Cincinnati,OH，美国)，溶于每加仑含有3格令混合Ca2+/Mg2+硬度的溶液中，20℃下每孔使用0.3ppm的酶。
表6
| A | B | C | D | |||||||||||||
| N76D | S103A | V104I | 1 | 1 | 1 | 1 | ||||||||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.44 | 1.41 | 1.39 | 1.26 | |||||
| V68A | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | 2.34 | 1.49 | 1.65 | 2.35 | |||
| V68A | S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.05 | 1.41 | 1.20 | 1.19 | |||
| G20R | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.81 | 1.72 | 1.66 | 1.31 | |||
| V68A | S103A | V104I | G159D | Y209F | A232V | Q236H | Q245R | N248D | N252K | 2.19 | 1.38 | 1.60 | 2.02 | |||
| V68A | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | 2.91 | 0.91 | 1.48 | 2.70 | |||
| V68A | S103A | V104I | G159D | P210R | A232V | Q236H | Q245R | N248D | N252K | 0.93 | 1.39 | 1.23 | 0.80 | |||
| V68A | S103A | V104I | G159D | N185D | P210L | A232V | Q236H | Q245R | N248D | N252K | 2.67 | 0.86 | 1.41 | 2.88 | ||
| V68A | S103A | V104I | G159D | P210L | A232V | Q236H | Q245R | N248D | N252K | 2.22 | 1.43 | 1.55 | 1.78 | |||
| V68A | S103A | V104I | G159D | S212C | A232V | Q236H | Q245R | N248D | N252K | 2.30 | 1.43 | 1.63 | 2.07 | |||
| V68A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 2.31 | 1.47 | 1.62 | 2.01 | |||
| V68A | S103A | V104I | G159D | S212E | A232V | Q236H | Q245R | N248D | N252K | 2.63 | 0.56 | 1.36 | 2.66 | |||
| V68A | S103A | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | 2.75 | 0.50 | 1.27 | 2.78 | |||
| V68A | S103A | V104I | T213S | A232V | Q236H | Q245R | N248D | N252K | 1.11 | 1.38 | 1.31 | 0.75 |
表6(续)
| V68A | A103V | V104I | G159D | T213E | A232V | Q236H | Q245R | N248D | N252K | 2.27 | 0.15 | 1.12 | 2.01 | |||
| V68A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 1.37 | 1.42 | 1.37 | 1.06 | |||
| V68A | S103A | V104I | G159D | A215V | A232V | Q236H | Q245R | N248D | N252K | 2.14 | 1.40 | 1.53 | 1.54 | |||
| V68A | S103A | V104I | G159D | A215R | A232V | Q236H | Q245R | N248D | N252K | 1.22 | 1.58 | 1.47 | 1.20 | |||
| V68A | S103A | V104I | G159D | S216T | A232V | Q236H | Q245R | N248D | N252K | 2.12 | 1.36 | 1.56 | 1.56 | |||
| V68A | S103A | V104I | G159D | S216V | A232V | Q236H | Q245R | N248D | N252K | 1.88 | 1.36 | 1.47 | 1.87 | |||
| V68A | S103A | V104I | G159D | S216C | A232V | Q236H | Q245R | N248D | N252K | 2.24 | 0.33 | 1.07 | 2.89 | |||
| V68A | S103A | V104I | G159D | N173D | A232V | Q236H | Q245R | N248D | N252K | 2.43 | 0.46 | 1.29 | 2.42 | |||
| V68A | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | 0.98 | 1.46 | 1.24 | 0.95 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F | 2.52 | 1.00 | 1.42 | 2.42 | ||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252L | 2.05 | 1.13 | 1.30 | 1.85 | ||||
| P55S | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252F | 2.61 | 0.91 | 1.43 | 3.22 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T255V | 2.18 | 1.36 | 1.58 | 1.72 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256N | 2.14 | 1.46 | 1.59 | 1.65 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256E | 2.46 | 0.77 | 1.33 | 2.58 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | S256R | 1.31 | 1.52 | 1.46 | 0.94 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | T260R | 1.21 | 1.41 | 1.31 | 1.05 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | L257R | 1.51 | 1.41 | 0.85 | 1.18 |
表6(续)
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | G258D | 2.56 | 0.59 | 1.30 | 2.64 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | 1.02 | 1.47 | 1.37 | 0.84 | |||
| V68A | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | 1.04 | 1.50 | 1.32 | 0.73 | |||||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245V | N248D | N252K | 2.60 | 0.93 | 1.41 | 2.67 | ||||
| V68A | S103A | V104I | G159D | A228V | A232V | Q236H | Q245R | N248D | N252K | 2.31 | 1.38 | 1.53 | 1.57 | |||
| G61E | V68A | S103A | V104I | S130A | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.83 | 0.25 | 1.33 | 2.44 | ||
| G61E | S103A | V104I | A133V | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.10 | 0.97 | 1.36 | 2.29 | |||
| V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248G | N252K | 1.37 | 1.54 | 0.89 | 1.27 | ||||
| V68A | S103A | V104I | G159D | N218S | A232V | Q236H | Q245R | N248D | N252K | 2.30 | 1.50 | 1.62 | 1.56 | |||
| G20R | V68A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.72 | 1.72 | 1.67 | 1.15 | |||
| V68A | N76D | E89D | S103A | V104I | G159D | P210L | T213R | A232V | Q236H | Q245R | T260A | 1.32 | 1.30 | 1.11 | 1.28 | |
| V68A | N76D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.50 | 0.83 | 1.43 | 2.25 | |||
| G61E | V68A | S103A | V104I | G159D | S160V | A232V | Q236H | Q245R | N248D | N252K | 4.20 | 0.07 | ND | 1.28 | ||
| S3L | G61E | V68A | N76D | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | 3.47 | 0.60 | ND | 1.45 | ||
| G61E | V68A | S103A | V104I | G159D | Y167F | A232V | Q236H | Q245R | N248D | N252K | 4.32 | 0.79 | ND | 1.55 | ||
| G97E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.14 | 0.41 | ND | 1.40 | ||||
| A98D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.71 | 0.68 | ND | 1.72 | ||||
| S99E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.97 | 0.68 | ND | 1.71 |
表6(续)
| S101E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.50 | 0.27 | ND | 1.90 | ||||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.24 | 1.80 | ND | 1.33 | ||||
| G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.35 | 1.33 | ND | 1.69 | ||||
| S103A | V104I | S106E | G159D | A232V | Q236H | Q245R | N248D | N252K | 4.88 | 0.55 | ND | 2.71 | ||||
| S103A | V104I | Q109E | G159D | A232V | Q236H | Q245R | N248D | N252K | 4.22 | 1.05 | ND | 2.40 | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | 5.45 | 2.19 | ND | 2.58 | ||||
| S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | 3.76 | 2.16 | ND | 1.82 | ||||
| N62D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 7.42 | 0.13 | ND | 2.46 | ||||
| S103A | V104I | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | 5.43 | 1.36 | ND | 2.84 | ||||
| S103A | V104I | G159D | S166D | A232V | Q236H | Q245R | N248D | N252K | 5.12 | 1.21 | ND | 3.97 | ||||
| S103A | V104I | G159D | L217E | A232V | Q236H | Q245R | N248D | N252K | 6.38 | 0.95 | ND | 3.09 | ||||
| G20R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 3.17 | 2.83 | ND | 2.60 | ||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 4.38 | 1.92 | ND | 2.54 | |||
| S103A | V104I | G159D | Q206R | L217E | A232V | Q236H | Q245R | N248D | N252K | 3.05 | 2.61 | ND | 1.10 | |||
| N62D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | 4.09 | 2.46 | ND | 2.55 | |||
| S103A | V104I | G159D | N184G | A232V | Q236H | Q245R | N248D | N252K | 2.32 | 2.08 | ND | 2.40 | ||||
| S103A | V104I | G159D | A232V | Q236H | V244T | Q245R | N248D | N252K | 2.34 | 2.04 | ND | 1.86 | ||||
| S103A | V104I | G159D | A232V | Q236H | V244A | Q245R | N248D | N252K | 2.24 | 2.11 | ND | 1.95 |
表6(续)
| K27N | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.81 | 1.56 | ND | 1.82 | ||||
| T38G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.30 | 2.09 | ND | 1.82 | ||||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | S256R | 2.63 | 2.66 | ND | 1.44 | ||
| Q12R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 2.01 | 2.78 | ND | 1.99 | ||
| N62D | S103A | V104I | G159D | N185D | Q206E | T213R | A232V | Q236H | Q245R | N248D | N252K | E271Q | 7.74 | 0.94 | ND | 5.39 |
| S101G | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | 5.14 | 1.41 | ND | 1.92 | |||
| S101G | S103A | V104I | G159D | Q206E | A232V | Q236H | Q245R | N248D | N252K | 4.97 | 0.57 | ND | 1.36 | |||
| S101G | S103A | V104I | G159D | T213Q | A232V | Q236H | Q245R | N248D | N252K | 2.41 | 1.86 | ND | 1.01 | |||
| A98L | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 4.42 | 0.50 | ND | 2.88 | |||
| S101G | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 5.86 | 1.20 | ND | 3.84 | |||
| G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 5.87 | 2.10 | ND | 3.19 | |||
| Q12R | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 2.98 | 2.67 | ND | 2.17 | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 4.02 | 0.41 | ND | 2.25 | ||
| S101G | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 6.63 | 2.07 | ND | 2.08 | ||
| G102A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 2.03 | 2.48 | ND | 2.25 | |||
| N62D | S103A | V104I | Q109R | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 2.96 | 2.76 | ND | 2.34 | ||
| S103A | V104I | G159D | A232V | Q245R | N248D | N252K | 2.74 | 2.10 | ND | 1.86 | ||||||
| S103A | V104I | G159D | A230V | Q245R | 2.11 | 2.35 | ND | 1.49 |
表6(续)
表6(续)
表6(续)
表6(续)
表6(续)
| N62D | S103A | V104I | S130G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 3.42 | 0.71 | ND | 2.58 | ||
| S101G | S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.59 | 1.32 | ND | 1.61 | |||
| S101G | S103A | V104I | S128G | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.30 | 1.23 | ND | 9.0 | |||
| S101G | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.94 | 0.71 | ND | 1.08 | |||
| N62D | S101G | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 3.17 | 0.83 | ND | 2.35 | ||
| N62D | S103A | V104I | S128G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 2.15 | 1.38 | ND | 1.77 | ||
| N62D | S103A | V104I | S128L | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 3.07 | 0.07 | ND | 1.45 | ||
| S101G | S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.26 | 1.16 | ND | 3.05 | |||
| A98V | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 1.82 | 1.34 | ND | 1.08 | |||
| S99G | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 2.16 | 1.47 | ND | 1.20 | |||
| S101G | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 1.79 | 1.38 | ND | 1.01 | |||
| S101G | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | 1.15 | 1.18 | ND | 8.7 | |||
| S101G | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | 1.47 | 1.23 | ND | 1.03 | |||
| S101G | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | N248D | N252K | 1.90 | 1.38 | ND | 1.05 | |||
| S101G | S103A | V104I | G159D | A230V | Q236H | Q245R | 1.55 | 1.51 | ND | 1.23 | ||||||
| S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | 1.96 | 1.30 | ND | 1.10 | |||
| N76D | S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | 2.49 | 0.80 | ND | 1.25 | ||
| G61E | V68A | S103A | V104I | G159D | S160V | A232V | Q236H | Q245R | N248D | N252K | 420 | 7 | 128 | ND |
| S3L | G61E | V68A | N76D | S103A | V104I | A232V | Q236H | Q245R | N248D | N252K | 347 | 60 | 145 | ND | ||
| G61E | V68A | S103A | V104I | G159D | Y167F | A232V | Q236H | Q245R | N248D | N252K | 432 | 79 | 155 | ND | ||
| G97E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 314 | 41 | 140 | ND | ||||
| A98D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 271 | 68 | 172 | ND | ||||
| S99E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 297 | 68 | 171 | ND | ||||
| S101E | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 350 | 27 | 190 | ND | ||||
| S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 224 | 180 | 133 | ND | ||||
| G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 335 | 133 | 169 | ND | ||||
| S103A | V104I | S106E | G159D | A232V | Q236H | Q245R | N248D | N252K | 488 | 55 | 271 | ND | ||||
| S103A | V104I | Q109E | G159D | A232V | Q236H | Q245R | N248D | N252K | 422 | 105 | 240 | ND | ||||
| S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | N261R | 545 | 219 | 258 | ND | ||||
| S103A | V104I | Q109R | G159D | A232V | Q236H | Q245R | N248D | N252K | 376 | 216 | 182 | ND | ||||
| N62D | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 742 | 13 | 246 | ND | ||||
| S103A | V104I | G159D | N184D | A232V | Q236H | Q245R | N248D | N252K | 543 | 136 | 284 | ND | ||||
| S103A | V104I | G159D | S166D | A232V | Q236H | Q245R | N248D | N252K | 512 | 121 | 397 | ND | ||||
| S103A | V104I | G159D | L217E | A232V | Q236H | Q245R | N248D | N252K | 638 | 95 | 309 | ND | ||||
| G20R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 317 | 283 | 260 | ND | ||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 438 | 192 | 254 | ND |
| S103A | V104I | G159D | Q206R | L217E | A232V | Q236H | Q245R | N248D | N252K | 305 | 261 | 110 | ND | |||
| N62D | S103A | V104I | G159D | Q206R | A232V | Q236H | Q245R | N248D | N252K | 409 | 246 | 255 | ND | |||
| S103A | V104I | G159D | N184G | A232V | Q236H | Q245R | N248D | N252K | 232 | 208 | 240 | ND | ||||
| S103A | V104I | G159D | A232V | Q236H | V244T | Q245R | N248D | N252K | 234 | 204 | 186 | ND | ||||
| S103A | V104I | G159D | A232V | Q236H | V244A | Q245R | N248D | N252K | 224 | 211 | 195 | ND | ||||
| K27N | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 281 | 156 | 247 | ND | ||||
| T38G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 230 | 209 | 182 | ND | ||||
| N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | S256R | 263 | 266 | 144 | ND | ||
| Q12R | N62D | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 201 | 278 | 199 | |||
| N62D | S103A | V104I | G159D | N185D | Q206E | T213R | A232V | Q236H | Q245R | N248D | N252K | E271Q | 774 | 94 | 539 | ND |
| S101G | S103A | V104I | G159D | N185D | A232V | Q236H | Q245R | N248D | N252K | 514 | 141 | 192 | ND | |||
| S101G | S103A | V104I | G159D | Q206E | A232V | Q236H | Q245R | N248D | N252K | 497 | 57 | 136 | ND | |||
| S101G | S103A | V104I | G159D | T213Q | A232V | Q236H | Q245R | N248D | N252K | 241 | 186 | 101 | ND | |||
| A98L | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 442 | 50 | 288 | ND | |||
| S101G | G102A | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 586 | 120 | 384 | ND | |||
| G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 587 | 210 | 319 | ND | |||
| Q12R | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 298 | 267 | 217 | ND | ||
| A98L | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 402 | 41 | 225 | ND |
| S101G | G102A | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 663 | 207 | 208 | ND | ||
| G102A | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 203 | 248 | 225 | ND | |||
| N62D | S103A | V104I | Q109R | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 296 | 276 | 234 | ND | ||
| S103A | V104I | G159D | A232V | Q245R | N248D | N252K | 274 | 210 | 186 | ND | ||||||
| S103A | V104I | G159D | A230V | Q245R | 211 | 235 | 149 | ND | ||||||||
| N62D | S103A | V104I | S130G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 342 | 71 | 258 | ND | ||
| S101G | S103A | V104I | S130G | G159D | A232V | Q236H | Q245R | N248D | N252K | 259 | 132 | 161 | ND | |||
| S101G | S103A | V104I | S128G | G159D | A232V | Q236H | Q245R | N248D | N252K | 130 | 123 | 90 | ND | |||
| S101G | S103A | V104I | S128L | G159D | A232V | Q236H | Q245R | N248D | N252K | 294 | 71 | 108 | ND | |||
| N62D | S101G | S103A | V104I | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 317 | 83 | 235 | ND | ||
| N62D | S103A | V104I | S128G | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 215 | 138 | 177 | ND | ||
| N62D | S103A | V104I | S128L | G159D | T213R | A232V | Q236H | Q245R | N248D | N252K | 307 | 7 | 145 | ND | ||
| S101G | S103A | V104I | P131V | G159D | A232V | Q236H | Q245R | N248D | N252K | 226 | 116 | 305 | ND | |||
| A98V | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 182 | 134 | 108 | ND | |||
| S99G | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 216 | 147 | 120 | ND | |||
| S101G | S103A | V104I | G159D | S212G | A232V | Q236H | Q245R | N248D | N252K | 179 | 138 | 101 | ND | |||
| S101G | S103A | V104I | G159D | Y209W | A232V | Q236H | Q245R | N248D | N252K | 115 | 118 | 87 | ND | |||
| S101G | S103A | V104I | G159D | P210I | A232V | Q236H | Q245R | N248D | N252K | 147 | 123 | 103 | ND |
| S101G | S103A | V104I | G159D | V205I | A232V | Q236H | Q245R | N248D | N252K | 190 | 138 | 105 | ND | |||
| S101G | S103A | V104I | G159D | A230V | Q236H | Q245R | 155 | 151 | 123 | ND | ||||||
| S101G | S103A | V104I | G159D | A194P | A232V | Q236H | Q245R | N248D | N252K | 196 | 130 | 110 | ND | |||
| N76D | S101G | S103A | V104I | G159D | A232V | Q236H | Q245R | N248D | N252K | 249 | 80 | 125 | ND |
Claims (12)
1．一种蛋白酶变体，其包含在一个或多个残基位置处置换氨基酸，所述残基位置相应于选自解淀粉芽孢杆菌枯草杆菌蛋白酶62、212、230、232、252和257的残基位置。
2．根据权利要求1的蛋白酶变体，其来源于芽孢杆菌属的枯草杆菌蛋白酶。
3．根据权利要求2的蛋白酶变体，其来源于迟缓芽孢杆菌枯草杆菌蛋白酶。
4．一种编码权利要求1的蛋白酶变体的DNA。
5．一种编码权利要求4的DNA的表达载体。
6．一种用权利要求5的表达载体转化的宿主细胞。
7．一种含有权利要求1的蛋白酶变体的清洁用组合物。
8．一种含有权利要求1的蛋白酶变体的动物饲料。
9．一种含有权利要求1的蛋白酶变体的用于处理纺织品的组合物。
10．根据权利要求1的蛋白酶变体，其包含选自相应于表1中解淀粉芽孢杆菌枯草杆菌蛋白酶的位置的残基位置的置换。
11．根据权利要求10的蛋白酶变体，其包含选自相应于表2中解淀粉芽孢杆菌枯草杆菌蛋白酶的位置的残基位置的置换。
12．根据权利要求10的蛋白酶变体，其包含选自相应于表3中解淀粉芽孢杆菌枯草杆菌蛋白酶的位置的残基位置的置换。
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US95632397A | 1997-10-23 | 1997-10-23 | |
| US95632497A | 1997-10-23 | 1997-10-23 | |
| US95656497A | 1997-10-23 | 1997-10-23 | |
| US08/956,324 | 1997-10-23 | ||
| US08/956,323 | 1997-10-23 | ||
| US08/956,564 | 1997-10-23 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1306577A true CN1306577A (zh) | 2001-08-01 |
| CN100342016C CN100342016C (zh) | 2007-10-10 |
Family
ID=27420734
Family Applications (9)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB988126109A Expired - Lifetime CN1191347C (zh) | 1997-10-23 | 1998-10-23 | 含多取代蛋白酶变体的清洗组合物 |
| CNB988126370A Expired - Lifetime CN1198912C (zh) | 1997-10-23 | 1998-10-23 | 包含多取代的蛋白酶变体的漂白组合物 |
| CN98811435A Pending CN1279721A (zh) | 1997-10-23 | 1998-10-23 | 具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 |
| CN2010105436254A Pending CN102021159A (zh) | 1997-10-23 | 1998-10-23 | 具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 |
| CNB2004100399770A Expired - Lifetime CN1295313C (zh) | 1997-10-23 | 1998-10-23 | 含多取代蛋白酶变体的清洗组合物 |
| CNB988114364A Expired - Fee Related CN100342016C (zh) | 1997-10-23 | 1998-10-23 | 多点置换的蛋白酶变体 |
| CNB988113139A Expired - Fee Related CN1195857C (zh) | 1997-10-23 | 1998-10-23 | 多点置换的蛋白酶变体 |
| CNB988126400A Expired - Fee Related CN100558871C (zh) | 1997-10-23 | 1998-10-23 | 含有多取代的蛋白酶变体和淀粉酶变体的清洗组合物 |
| CNB2004100399766A Expired - Fee Related CN100506963C (zh) | 1997-10-23 | 1998-10-23 | 包含多取代的蛋白酶变体的漂白组合物 |
Family Applications Before (5)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB988126109A Expired - Lifetime CN1191347C (zh) | 1997-10-23 | 1998-10-23 | 含多取代蛋白酶变体的清洗组合物 |
| CNB988126370A Expired - Lifetime CN1198912C (zh) | 1997-10-23 | 1998-10-23 | 包含多取代的蛋白酶变体的漂白组合物 |
| CN98811435A Pending CN1279721A (zh) | 1997-10-23 | 1998-10-23 | 具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 |
| CN2010105436254A Pending CN102021159A (zh) | 1997-10-23 | 1998-10-23 | 具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 |
| CNB2004100399770A Expired - Lifetime CN1295313C (zh) | 1997-10-23 | 1998-10-23 | 含多取代蛋白酶变体的清洗组合物 |
Family Applications After (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB988113139A Expired - Fee Related CN1195857C (zh) | 1997-10-23 | 1998-10-23 | 多点置换的蛋白酶变体 |
| CNB988126400A Expired - Fee Related CN100558871C (zh) | 1997-10-23 | 1998-10-23 | 含有多取代的蛋白酶变体和淀粉酶变体的清洗组合物 |
| CNB2004100399766A Expired - Fee Related CN100506963C (zh) | 1997-10-23 | 1998-10-23 | 包含多取代的蛋白酶变体的漂白组合物 |
Country Status (26)
| Country | Link |
|---|---|
| US (5) | US6482628B1 (zh) |
| EP (10) | EP1025241B2 (zh) |
| JP (7) | JP2001520046A (zh) |
| KR (6) | KR20010024555A (zh) |
| CN (9) | CN1191347C (zh) |
| AR (10) | AR016969A1 (zh) |
| AT (9) | ATE513479T1 (zh) |
| AU (7) | AU1199699A (zh) |
| BR (6) | BR9813879A (zh) |
| CA (9) | CA2307638C (zh) |
| CZ (6) | CZ20001479A3 (zh) |
| DE (7) | DE69839677D1 (zh) |
| DK (9) | DK1025194T4 (zh) |
| EG (2) | EG21711A (zh) |
| ES (7) | ES2319470T5 (zh) |
| HK (1) | HK1039159B (zh) |
| HU (3) | HUP0103080A2 (zh) |
| ID (4) | ID28256A (zh) |
| MA (2) | MA24811A1 (zh) |
| NO (3) | NO327607B1 (zh) |
| NZ (3) | NZ520771A (zh) |
| PL (1) | PL191529B1 (zh) |
| PT (8) | PT1027418E (zh) |
| TR (8) | TR200001112T2 (zh) |
| TW (2) | TW585912B (zh) |
| WO (5) | WO1999020769A2 (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109312323A (zh) * | 2016-06-09 | 2019-02-05 | 花王株式会社 | 突变碱性蛋白酶 |
Families Citing this family (563)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MA23346A1 (fr) * | 1993-10-14 | 1995-04-01 | Genencor Int | Variantes de la subtilisine |
| EP1009815B1 (en) * | 1997-08-29 | 2008-01-30 | Novozymes A/S | Protease variants and compositions |
| ES2367505T3 (es) * | 1997-10-23 | 2011-11-04 | Danisco Us Inc. | Variantes de proteasa con sustituciones múltiples con carga neta alterada para usar en detergentes. |
| AR016969A1 (es) * | 1997-10-23 | 2001-08-01 | Procter & Gamble | VARIANTE DE PROTEASA, ADN, VECTOR DE EXPRESIoN, MICROORGANISMO HUESPED, COMPOSICIoN DE LIMPIEZA, ALIMENTO PARA ANIMALES Y COMPOSICIoN PARA TRATAR UN TEXTIL |
| DE69939538D1 (de) | 1998-07-02 | 2008-10-23 | Univ Toronto | Mit einer kohlenhydratgruppe chemisch modifizierte proteine |
| WO2000024924A2 (en) * | 1998-10-23 | 2000-05-04 | The Procter & Gamble Company | Methods for screening protease variants for use in detergent compositions |
| DE69940978D1 (de) | 1998-11-10 | 2009-07-23 | Univ Toronto | Chemisch modifizierte mutante serinhydrolasen |
| WO2000064485A2 (en) * | 1999-04-28 | 2000-11-02 | Genencor International, Inc. | Specifically targeted catalytic antagonists and uses thereof |
| US6627744B2 (en) | 1999-07-02 | 2003-09-30 | Genencor International, Inc. | Synthesis of glycodendrimer reagents |
| CA2379729A1 (en) * | 1999-07-22 | 2001-02-01 | The Procter & Gamble Company | Protease conjugates having sterically protected clip sites |
| AU6085800A (en) * | 1999-07-22 | 2001-02-13 | Procter & Gamble Company, The | Subtilisin protease variants having amino acid substitutions in defined epitope regions |
| BR9917481A (pt) * | 1999-09-09 | 2002-05-21 | Procter & Gamble | Composição detergente contendo uma protease |
| KR100387166B1 (ko) * | 1999-12-15 | 2003-06-12 | 닛뽄덴끼 가부시끼가이샤 | 유기 일렉트로 루미네선스 소자 |
| US7067049B1 (en) * | 2000-02-04 | 2006-06-27 | Exxonmobil Oil Corporation | Formulated lubricant oils containing high-performance base oils derived from highly paraffinic hydrocarbons |
| DE10007608A1 (de) * | 2000-02-18 | 2001-08-30 | Henkel Kgaa | Protease und Percarbonat enthaltende Wasch- und Reinigungsmittel |
| AU3724801A (en) * | 2000-03-03 | 2001-09-12 | Novozymes A/S | Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same |
| US6777218B1 (en) | 2000-03-14 | 2004-08-17 | Novozymes A/S | Subtilase enzymes having an improved wash performance on egg stains |
| US6902922B2 (en) * | 2000-04-03 | 2005-06-07 | Novozymes A/S | Subtilisin variants |
| EP2258852A1 (en) * | 2000-04-28 | 2010-12-08 | Novozymes A/S | Lipolytic enzyme variant |
| WO2002002725A1 (en) * | 2000-06-30 | 2002-01-10 | The Procter & Gamble Company | Detergent compositions comprising a cyclodextrin glucanotransferase enzyme |
| MXPA03000120A (es) * | 2000-06-30 | 2003-10-15 | Procter & Gamble | Composiciones detergentes que comprenden una enzima alfa-amilasa maltogenica. |
| US20020155574A1 (en) | 2000-08-01 | 2002-10-24 | Novozymes A/S | Alpha-amylase mutants with altered properties |
| US6893855B2 (en) * | 2000-10-13 | 2005-05-17 | Novozymes A/S | Subtilase variants |
| US6803222B2 (en) * | 2000-11-22 | 2004-10-12 | Kao Corporation | Alkaline proteases |
| US7303907B2 (en) * | 2001-01-08 | 2007-12-04 | Health Protection Agency | Degradation and detection of TSE infectivity |
| EP1241112A3 (en) | 2001-03-15 | 2003-02-26 | The Procter & Gamble Company | Flexible multiple compartment pouch |
| CN100489094C (zh) * | 2001-03-23 | 2009-05-20 | 金克克国际有限公司 | 具有改变的免疫原性反应的蛋白质及制备和使用该蛋白质的方法 |
| EP1423513B1 (en) | 2001-05-15 | 2009-11-25 | Novozymes A/S | Alpha-amylase variant with altered properties |
| DK200101090A (da) | 2001-07-12 | 2001-08-16 | Novozymes As | Subtilase variants |
| CA2472206C (en) * | 2001-12-31 | 2013-04-09 | Genencor International, Inc. | Proteases producing an altered immunogenic response and methods of making and using the same |
| WO2003062381A2 (en) * | 2002-01-16 | 2003-07-31 | Genencor International, Inc. | Multiply-substituted protease variants |
| CA2472723C (en) * | 2002-01-16 | 2013-12-17 | Genencor International, Inc. | Multiply-substituted protease variants |
| US7223386B2 (en) * | 2002-03-11 | 2007-05-29 | Dow Corning Corporation | Preparations for topical skin use and treatment |
| US20030180281A1 (en) * | 2002-03-11 | 2003-09-25 | Bott Richard R. | Preparations for topical skin use and treatment |
| EP3023501A1 (en) | 2002-03-29 | 2016-05-25 | Danisco US Inc. | Enhanced protein expression in bacillus |
| EP1545235B1 (en) | 2002-07-29 | 2013-11-06 | Aminotech AS | Method for production of peptides and amino acids from protein-comprising material of animal origin |
| US7082689B2 (en) * | 2002-10-11 | 2006-08-01 | Black & Decker Inc. | Keyless shoe lock for reciprocating saw |
| TWI319007B (en) * | 2002-11-06 | 2010-01-01 | Novozymes As | Subtilase variants |
| US7888093B2 (en) | 2002-11-06 | 2011-02-15 | Novozymes A/S | Subtilase variants |
| DE10260903A1 (de) * | 2002-12-20 | 2004-07-08 | Henkel Kgaa | Neue Perhydrolasen |
| US20050059567A1 (en) * | 2003-09-11 | 2005-03-17 | The Procter & Gamble Company | Methods of formulating enzyme cocktails, enzyme cocktails for the removal of egg-based and grass-based stains and/or soils, compositions and products comprising same |
| KR20130083461A (ko) | 2003-11-19 | 2013-07-22 | 더 프록터 앤드 갬블 캄파니 | 세린 프로테아제, 세린 효소들을 인코딩하는 핵산 및 이를 편입시킨 벡터 및 숙주 세포 |
| US7985569B2 (en) | 2003-11-19 | 2011-07-26 | Danisco Us Inc. | Cellulomonas 69B4 serine protease variants |
| CN1942584B (zh) | 2004-02-13 | 2011-07-27 | 诺维信公司 | 蛋白酶变体 |
| ATE454440T1 (de) | 2004-05-17 | 2010-01-15 | Procter & Gamble | Bleichmittel mit carbohydratoxidase |
| US20090060933A1 (en) * | 2004-06-14 | 2009-03-05 | Estell David A | Proteases producing an altered immunogenic response and methods of making and using the same |
| JP2006143853A (ja) * | 2004-11-18 | 2006-06-08 | Lion Corp | アミラーゼ含有漂白性組成物 |
| JP2006143855A (ja) * | 2004-11-18 | 2006-06-08 | Lion Corp | 飲料シミ汚れ除去効果を向上させたアミラーゼ含有漂白性組成物 |
| US8814861B2 (en) | 2005-05-12 | 2014-08-26 | Innovatech, Llc | Electrosurgical electrode and method of manufacturing same |
| US7147634B2 (en) | 2005-05-12 | 2006-12-12 | Orion Industries, Ltd. | Electrosurgical electrode and method of manufacturing same |
| EP2385112B1 (en) * | 2005-07-08 | 2016-11-30 | Novozymes A/S | Subtilase variants |
| US20070161531A1 (en) * | 2005-07-08 | 2007-07-12 | Novozymes A/S | Subtilase variants |
| GB0603775D0 (en) * | 2006-02-24 | 2006-04-05 | Health Prot Agency | Infectivity assay |
| CN101415482B (zh) | 2006-03-31 | 2015-04-15 | 金克克国际有限公司 | 用于化合物分离的切向流动过滤设备、系统和方法 |
| US7629158B2 (en) | 2006-06-16 | 2009-12-08 | The Procter & Gamble Company | Cleaning and/or treatment compositions |
| CN101473036B (zh) | 2006-06-23 | 2012-12-12 | 丹尼斯科美国公司 | 利用用于工程改造多种性质的位点评价文库对序列和活性关系进行系统评价 |
| KR20150115023A (ko) * | 2006-07-18 | 2015-10-13 | 다니스코 유에스 인크. | 광범위한 온도에 걸쳐 활성인 프로테아제 변이형 |
| CA2660645C (en) | 2006-08-11 | 2016-04-05 | Novozymes Biologicals, Inc. | Bacillus cultures for use in washing, cleaning, stain removal, or degrading waste materials |
| US20100105598A1 (en) * | 2006-10-16 | 2010-04-29 | Pieter Augustinus | Non-Phosphate Dish Detergents |
| AU2008226792B2 (en) | 2007-03-12 | 2013-06-06 | Danisco Us Inc. | Modified proteases |
| US7922970B2 (en) * | 2007-04-03 | 2011-04-12 | Kimberly-Clark Worldwide, Inc. | Use of sonication to eliminate prions |
| CN101679489B (zh) | 2007-05-10 | 2014-05-28 | 丹尼斯科美国公司 | 增加细菌内多肽表达的修饰的分泌系统 |
| CA2690032C (en) * | 2007-06-06 | 2018-05-15 | Danisco Us Inc. | Methods for improving protein properties |
| DK2641652T3 (en) * | 2007-09-12 | 2019-04-29 | Danisco Us Inc | FILTERING WITH INTERNAL POLLUTION CONTROL |
| US8021436B2 (en) | 2007-09-27 | 2011-09-20 | The Procter & Gamble Company | Cleaning and/or treatment compositions comprising a xyloglucan conjugate |
| US7618801B2 (en) | 2007-10-30 | 2009-11-17 | Danison US Inc. | Streptomyces protease |
| JP5520828B2 (ja) | 2007-11-05 | 2014-06-11 | ダニスコ・ユーエス・インク | 改変される特徴を有するバシルス種（Ｂａｃｉｌｌｕｓｓｐ．）ＴＳ‐２３アルファ‐アミラーゼ変異体 |
| CN101932703B (zh) | 2007-11-05 | 2015-08-05 | 丹尼斯科美国公司 | 具有改变的特性的α-淀粉酶变体 |
| AU2008348270A1 (en) | 2007-12-21 | 2009-07-30 | Danisco Us Inc. | Enhanced protein production in bacillus |
| JP2011510681A (ja) | 2008-02-04 | 2011-04-07 | ダニスコ・ユーエス・インク | 改変された特性をもつｔｓ２３アルファ‐アミラーゼ変異体 |
| US8066818B2 (en) | 2008-02-08 | 2011-11-29 | The Procter & Gamble Company | Water-soluble pouch |
| PL2088187T3 (pl) * | 2008-02-08 | 2013-01-31 | Procter & Gamble | Sposób otrzymywania rozpuszczalnej w wodzie saszetki |
| US20090209447A1 (en) * | 2008-02-15 | 2009-08-20 | Michelle Meek | Cleaning compositions |
| EP2100948A1 (en) * | 2008-03-14 | 2009-09-16 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| US20090233830A1 (en) * | 2008-03-14 | 2009-09-17 | Penny Sue Dirr | Automatic detergent dishwashing composition |
| EP2100947A1 (en) | 2008-03-14 | 2009-09-16 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| CN107090014B (zh) | 2008-03-26 | 2023-04-25 | 诺维信公司 | 稳定化的液体酶组合物 |
| CN101981193B (zh) | 2008-03-28 | 2016-05-11 | 丹尼斯科美国公司 | 在细菌细胞中扩增基因座的方法 |
| EP2297314B1 (en) | 2008-06-06 | 2015-04-15 | Danisco US Inc. | Compositions and methods comprising variant microbial proteases |
| CN105483099B (zh) | 2008-06-06 | 2020-06-23 | 丹尼斯科美国公司 | 具有改良特性的嗜热脂肪土芽孢杆菌α-淀粉酶(AMYS)变体 |
| EP2166092A1 (en) | 2008-09-18 | 2010-03-24 | The Procter and Gamble Company | Detergent composition |
| EP2166073A1 (en) | 2008-09-23 | 2010-03-24 | The Procter & Gamble Company | Cleaning composition |
| EP2166076A1 (en) * | 2008-09-23 | 2010-03-24 | The Procter & Gamble Company | Cleaning composition |
| EP2166075A1 (en) | 2008-09-23 | 2010-03-24 | The Procter and Gamble Company | Cleaning composition |
| BRPI0920891B1 (pt) | 2008-09-25 | 2023-01-10 | Danisco Us Inc | Mistura de alfa-amilase e método para produção de um açúcar fermentável |
| JP5727378B2 (ja) * | 2008-11-11 | 2015-06-03 | ダニスコ・ユーエス・インク | セリンプロテアーゼ変異体を含む組成物及び方法 |
| EP2362902B1 (en) * | 2008-11-11 | 2012-10-24 | Danisco US, Inc., Genencor Division | Compositions and methods comprising a subtilisin variant |
| MX2011004802A (es) | 2008-11-11 | 2011-06-16 | Danisco Inc | Bacilos de subtilisina que comprenden una o mas mutaciones combinables. |
| US8183024B2 (en) | 2008-11-11 | 2012-05-22 | Danisco Us Inc. | Compositions and methods comprising a subtilisin variant |
| US20100267304A1 (en) * | 2008-11-14 | 2010-10-21 | Gregory Fowler | Polyurethane foam pad and methods of making and using same |
| US20100125046A1 (en) | 2008-11-20 | 2010-05-20 | Denome Frank William | Cleaning products |
| EP2216391A1 (en) | 2009-02-02 | 2010-08-11 | The Procter & Gamble Company | Liquid hand dishwashing detergent composition |
| ES2488117T3 (es) | 2009-02-02 | 2014-08-26 | The Procter & Gamble Company | Composición detergente líquida para lavado de vajillas a mano |
| PL2213713T3 (pl) | 2009-02-02 | 2014-07-31 | Procter & Gamble | Płynna kompozycja środka czyszczącego do ręcznego mycia naczyń |
| EP2216392B1 (en) | 2009-02-02 | 2013-11-13 | The Procter and Gamble Company | Liquid hand dishwashing detergent composition |
| EP2216390B1 (en) | 2009-02-02 | 2013-11-27 | The Procter and Gamble Company | Hand dishwashing method |
| EP2213715A1 (en) * | 2009-02-02 | 2010-08-04 | The Procter & Gamble Company | Liquid hand dishwashing detergent composition |
| EP2216393B1 (en) | 2009-02-09 | 2024-04-24 | The Procter & Gamble Company | Detergent composition |
| US8153574B2 (en) | 2009-03-18 | 2012-04-10 | The Procter & Gamble Company | Structured fluid detergent compositions comprising dibenzylidene polyol acetal derivatives and detersive enzymes |
| US8293697B2 (en) | 2009-03-18 | 2012-10-23 | The Procter & Gamble Company | Structured fluid detergent compositions comprising dibenzylidene sorbitol acetal derivatives |
| US8852912B2 (en) | 2009-04-01 | 2014-10-07 | Danisco Us Inc. | Compositions and methods comprising alpha-amylase variants with altered properties |
| WO2010138347A1 (en) | 2009-05-26 | 2010-12-02 | The Procter & Gamble Company | Aqueous liquid composition for pre-treating soiled dishware |
| AR076941A1 (es) | 2009-06-11 | 2011-07-20 | Danisco Us Inc | Cepa de bacillus para una mayor produccion de proteina |
| WO2011005623A1 (en) | 2009-07-09 | 2011-01-13 | The Procter & Gamble Company | Laundry detergent composition comprising low level of bleach |
| EP2451922A1 (en) | 2009-07-09 | 2012-05-16 | The Procter & Gamble Company | Method of laundering fabric using a compacted liquid laundry detergent composition |
| US20110009307A1 (en) | 2009-07-09 | 2011-01-13 | Alan Thomas Brooker | Laundry Detergent Composition Comprising Low Level of Sulphate |
| BR112012000531A2 (pt) | 2009-07-09 | 2019-09-24 | Procter & Gamble | composição detergente para lavagem de roupas catalítica que compreende teores relativamente baixos de eletrólito solúvel em água |
| WO2011005730A1 (en) | 2009-07-09 | 2011-01-13 | The Procter & Gamble Company | A catalytic laundry detergent composition comprising relatively low levels of water-soluble electrolyte |
| EP2451923A1 (en) | 2009-07-09 | 2012-05-16 | The Procter & Gamble Company | Method of laundering fabric using a liquid laundry detergent composition |
| WO2011005804A1 (en) | 2009-07-09 | 2011-01-13 | The Procter & Gamble Company | Method of laundering fabric using a liquid laundry detergent composition |
| MX342487B (es) | 2009-07-09 | 2016-09-29 | The Procter & Gamble Company * | Composicion detergente solida para tratamiento de tela con bajo contenido de aditivo. ligeramente alcalina, que comprende acido ftalimido peroxicaproico. |
| US20130137799A1 (en) | 2009-07-31 | 2013-05-30 | Akzo Nobel N.V. | Graft copolymers |
| ES2581916T5 (es) | 2009-08-13 | 2022-11-07 | Procter & Gamble | Método para lavado de tejidos a baja temperatura |
| EP2480649A1 (en) * | 2009-09-25 | 2012-08-01 | Novozymes A/S | Detergent composition |
| MX339581B (es) * | 2009-09-25 | 2016-06-01 | Novozymes As | Variantes de subtilasa. |
| US8785171B2 (en) | 2009-12-09 | 2014-07-22 | The Procter & Gamble Company | Fabric and home care products comprising cold water proteases |
| JP5863666B2 (ja) | 2009-12-09 | 2016-02-17 | ダニスコ・ユーエス・インク | プロテアーゼ変異体を含む組成物及び方法 |
| PL2333042T3 (pl) | 2009-12-10 | 2015-12-31 | Procter & Gamble | Produkt do automatycznych zmywarek do mycia naczyń i jego wykorzystanie |
| EP2333041B1 (en) | 2009-12-10 | 2013-05-15 | The Procter & Gamble Company | Method and use of a dishwasher composition |
| ES2399311T5 (es) | 2009-12-10 | 2020-06-19 | Procter & Gamble | Composición detergente |
| ES2423580T5 (es) | 2009-12-10 | 2021-06-17 | Procter & Gamble | Método y uso de una composición para lavado de vajillas |
| US20120258900A1 (en) | 2009-12-21 | 2012-10-11 | Danisco Us Inc. | Detergent compositions containing bacillus subtilis lipase and methods of use thereof |
| US8741609B2 (en) | 2009-12-21 | 2014-06-03 | Danisco Us Inc. | Detergent compositions containing Geobacillus stearothermophilus lipase and methods of use thereof |
| EP2516610A1 (en) | 2009-12-21 | 2012-10-31 | Danisco US Inc. | Detergent compositions containing thermobifida fusca lipase and methods of use thereof |
| CA2785624A1 (en) * | 2010-01-22 | 2011-07-28 | Dupont Nutrition Biosciences Aps | Methods for producing amino-substituted glycolipid compounds |
| EP3404087A1 (en) | 2010-02-10 | 2018-11-21 | Novozymes A/S | Alpha-amylase variants with high stability in presence of a chelating agent |
| EP2357220A1 (en) | 2010-02-10 | 2011-08-17 | The Procter & Gamble Company | Cleaning composition comprising amylase variants with high stability in the presence of a chelating agent |
| EP2361964B1 (en) | 2010-02-25 | 2012-12-12 | The Procter & Gamble Company | Detergent composition |
| EP2365059A1 (en) | 2010-03-01 | 2011-09-14 | The Procter & Gamble Company | Solid laundry detergent composition comprising C.I. fluorescent brightener 260 in alpha-crystalline form |
| EP2377914B1 (en) | 2010-04-19 | 2016-11-09 | The Procter & Gamble Company | Mildly alkaline, low-built, solid fabric treatment detergent composition comprising perhydrolase |
| EP2363456A1 (en) | 2010-03-01 | 2011-09-07 | The Procter & Gamble Company | Solid laundry detergent composition comprising brightener in micronized particulate form |
| EP2365054A1 (en) | 2010-03-01 | 2011-09-14 | The Procter & Gamble Company | Solid laundry detergent composition comprising secondary alcohol-based detersive surfactant |
| EP2380957A1 (en) | 2010-04-19 | 2011-10-26 | The Procter & Gamble Company | Solid laundry detergent composition having a dynamic in-wash ph profile |
| EP2365058A1 (en) | 2010-03-01 | 2011-09-14 | The Procter & Gamble Company | Solid laundry detergent composition having an excellent anti-encrustation profile |
| AR080886A1 (es) | 2010-04-15 | 2012-05-16 | Danisco Us Inc | Composiciones y metodos que comprenden proteasas variantes |
| US20120067373A1 (en) | 2010-04-15 | 2012-03-22 | Philip Frank Souter | Automatic Dishwashing Detergent Composition |
| US20110257062A1 (en) | 2010-04-19 | 2011-10-20 | Robert Richard Dykstra | Liquid laundry detergent composition comprising a source of peracid and having a ph profile that is controlled with respect to the pka of the source of peracid |
| US8889612B2 (en) | 2010-04-19 | 2014-11-18 | The Procter & Gamble Company | Method of laundering fabric using a compacted liquid laundry detergent composition |
| US20110257060A1 (en) | 2010-04-19 | 2011-10-20 | Robert Richard Dykstra | Laundry detergent composition comprising bleach particles that are suspended within a continuous liquid phase |
| US20110257069A1 (en) | 2010-04-19 | 2011-10-20 | Stephen Joseph Hodson | Detergent composition |
| EP2380478A1 (en) | 2010-04-23 | 2011-10-26 | The Procter & Gamble Company | Automatic dishwashing product |
| EP2383329A1 (en) | 2010-04-23 | 2011-11-02 | The Procter & Gamble Company | Particle |
| PL2380963T3 (pl) | 2010-04-23 | 2016-07-29 | Procter & Gamble | Sposób perfumowania |
| EP2380481B1 (en) | 2010-04-23 | 2014-12-31 | The Procter and Gamble Company | Automatic dishwashing product |
| ES2579217T3 (es) | 2010-04-23 | 2016-08-08 | The Procter & Gamble Company | Partícula |
| TR201810936T4 (tr) | 2010-04-23 | 2018-08-27 | Industrial Chemicals Group Ltd | Deterjan bileşimi. |
| US20130260438A1 (en) * | 2010-05-06 | 2013-10-03 | Danisco Us Inc. | Compositions and methods comprising serine protease variants (as amended) |
| DE102010028951A1 (de) | 2010-05-12 | 2011-11-17 | Henkel Ag & Co. Kgaa | Lagerstabiles flüssiges Wasch- oder Reinigungsmittel enthaltend Protease und Lipase |
| WO2011150157A2 (en) | 2010-05-28 | 2011-12-01 | Danisco Us Inc. | Detergent compositions containing streptomyces griseus lipase and methods of use thereof |
| EP2395071A1 (en) | 2010-06-10 | 2011-12-14 | The Procter & Gamble Company | Solid detergent composition comprising lipase of bacterial origin |
| EP2395070A1 (en) | 2010-06-10 | 2011-12-14 | The Procter & Gamble Company | Liquid laundry detergent composition comprising lipase of bacterial origin |
| EP2585573A1 (en) | 2010-06-23 | 2013-05-01 | The Procter and Gamble Company | Product for pre-treatment and laundering of stained fabric |
| EP2412792A1 (en) | 2010-07-29 | 2012-02-01 | The Procter & Gamble Company | Liquid detergent composition |
| US8685171B2 (en) | 2010-07-29 | 2014-04-01 | The Procter & Gamble Company | Liquid detergent composition |
| CN101922108B (zh) * | 2010-09-14 | 2012-09-05 | 东华大学 | 使用1，4，7-三氮杂环壬烷配合物的活化漂白的方法 |
| WO2012057781A1 (en) | 2010-10-29 | 2012-05-03 | The Procter & Gamble Company | Cleaning and/or treatment compositions comprising a fungal serine protease |
| FI123942B (fi) | 2010-10-29 | 2013-12-31 | Ab Enzymes Oy | Sieniperäisen seriiniproteaasin variantteja |
| WO2011026154A2 (en) | 2010-10-29 | 2011-03-03 | The Procter & Gamble Company | Cleaning and/or treatment compositions |
| DE102010063458A1 (de) * | 2010-12-17 | 2012-06-21 | Henkel Ag & Co. Kgaa | Lagerstabiles flüssiges Wasch- oder Reinigungsmittel enthaltend Protease und Amylase |
| WO2012112718A1 (en) | 2011-02-15 | 2012-08-23 | Novozymes Biologicals, Inc. | Mitigation of odor in cleaning machines and cleaning processes |
| DE102011005354A1 (de) | 2011-03-10 | 2012-09-13 | Henkel Ag & Co. Kgaa | Leistungsverbesserte Proteasevarianten |
| US8802388B2 (en) | 2011-04-29 | 2014-08-12 | Danisco Us Inc. | Detergent compositions containing Bacillus agaradhaerens mannanase and methods of use thereof |
| AR086216A1 (es) | 2011-04-29 | 2013-11-27 | Danisco Us Inc | Composiciones detergentes que contienen mananasa de bacillus sp. y sus metodos de uso |
| BR112013027209A2 (pt) | 2011-04-29 | 2016-11-29 | Danisco Us Inc | "polipeptídeo recombinante, composição detergente contendo mananase de geobacillus tepidamans, método para limpeza de produto têxtil, vetor de expressão e célula hospedeira" |
| US9856466B2 (en) | 2011-05-05 | 2018-01-02 | Danisco Us Inc. | Compositions and methods comprising serine protease variants |
| RU2663114C2 (ru) | 2011-05-05 | 2018-08-01 | Дзе Проктер Энд Гэмбл Компани | Способы и композиции, содержащие варианты сериновой протеазы |
| US20140371435A9 (en) | 2011-06-03 | 2014-12-18 | Eduardo Torres | Laundry Care Compositions Containing Thiophene Azo Dyes |
| EP2537918A1 (en) | 2011-06-20 | 2012-12-26 | The Procter & Gamble Company | Consumer products with lipase comprising coated particles |
| US20120324655A1 (en) | 2011-06-23 | 2012-12-27 | Nalini Chawla | Product for pre-treatment and laundering of stained fabric |
| DK3421595T3 (da) | 2011-06-30 | 2020-10-26 | Novozymes As | Alfa-amylasevarianter |
| EP2540824A1 (en) | 2011-06-30 | 2013-01-02 | The Procter & Gamble Company | Cleaning compositions comprising amylase variants reference to a sequence listing |
| EP2551335A1 (en) | 2011-07-25 | 2013-01-30 | The Procter & Gamble Company | Enzyme stabilized liquid detergent composition |
| WO2013016030A1 (en) | 2011-07-27 | 2013-01-31 | The Procter & Gamble Company | Multiphase liquid detergent composition |
| US8841246B2 (en) | 2011-08-05 | 2014-09-23 | Ecolab Usa Inc. | Cleaning composition containing a polysaccharide hybrid polymer composition and methods of improving drainage |
| US8853144B2 (en) | 2011-08-05 | 2014-10-07 | Ecolab Usa Inc. | Cleaning composition containing a polysaccharide graft polymer composition and methods of improving drainage |
| US8636918B2 (en) | 2011-08-05 | 2014-01-28 | Ecolab Usa Inc. | Cleaning composition containing a polysaccharide hybrid polymer composition and methods of controlling hard water scale |
| US8679366B2 (en) | 2011-08-05 | 2014-03-25 | Ecolab Usa Inc. | Cleaning composition containing a polysaccharide graft polymer composition and methods of controlling hard water scale |
| BR112014004444A2 (pt) | 2011-08-31 | 2017-03-21 | Danisco Us Inc | composições e métodos que compreendem uma variante de enzima lipolítica |
| PL2584028T3 (pl) | 2011-10-19 | 2017-10-31 | Procter & Gamble | Cząstka |
| MX2014005094A (es) | 2011-11-04 | 2014-08-08 | Akzo Nobel Chemicals Int Bv | Copolimeros de dendrita hibridos, composiciones de los mismos y metodos para producirlos. |
| MX2014005089A (es) | 2011-11-04 | 2014-08-08 | Akzo Nobel Chemicals Int Bv | Copolimeros de dendrita de injerto, y metodos para producir los mismos. |
| JP6378089B2 (ja) | 2011-12-09 | 2018-08-22 | ダニスコ・ユーエス・インク | 微生物におけるタンパク質産生のための、Ｂ．ズブチリス（Ｂ．ｓｕｂｔｉｌｉｓ）からのリボソームプロモーター |
| BR112014012910B1 (pt) | 2011-12-13 | 2021-06-29 | Danisco Us Inc. | Método para produção de uma mistura de enzimas que catalisam um processo de conversão e método para preparação de um banco de células |
| WO2013092635A1 (en) * | 2011-12-20 | 2013-06-27 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| US20150017700A1 (en) | 2011-12-22 | 2015-01-15 | Danisco Us Inc. | Compositions and methods comprising a lipolytic enzyme variant |
| ES2644007T3 (es) | 2012-01-26 | 2017-11-27 | Novozymes A/S | Uso de polipéptidos con actividad de proteasa en piensos para animales y en detergentes |
| DK2623586T3 (da) | 2012-02-03 | 2017-11-13 | Procter & Gamble | Sammensætninger og fremgangsmåder til overfladebehandling med lipaser |
| TR201900214T4 (tr) | 2012-03-19 | 2019-02-21 | Milliken & Co | Karboksilat Boyalar |
| US9909109B2 (en) | 2012-04-02 | 2018-03-06 | Novozymes A/S | Lipase variants and polynucleotides encoding same |
| WO2013167581A1 (en) | 2012-05-07 | 2013-11-14 | Novozymes A/S | Polypeptides having xanthan degrading activity and polynucleotides encoding same |
| WO2013178670A2 (de) * | 2012-05-30 | 2013-12-05 | Clariant International Ltd. | Verwendung von n-methyl-n-acylglucaminen als kältestabilisatoren in tensidlösungen |
| EP2674475A1 (en) | 2012-06-11 | 2013-12-18 | The Procter & Gamble Company | Detergent composition |
| AU2013279440B2 (en) | 2012-06-20 | 2016-10-06 | Novozymes A/S | Use of polypeptides having protease activity in animal feed and detergents |
| WO2014009473A1 (en) | 2012-07-12 | 2014-01-16 | Novozymes A/S | Polypeptides having lipase activity and polynucleotides encoding same |
| US8945314B2 (en) | 2012-07-30 | 2015-02-03 | Ecolab Usa Inc. | Biodegradable stability binding agent for a solid detergent |
| US8871699B2 (en) | 2012-09-13 | 2014-10-28 | Ecolab Usa Inc. | Detergent composition comprising phosphinosuccinic acid adducts and methods of use |
| US20140308162A1 (en) | 2013-04-15 | 2014-10-16 | Ecolab Usa Inc. | Peroxycarboxylic acid based sanitizing rinse additives for use in ware washing |
| US9994799B2 (en) | 2012-09-13 | 2018-06-12 | Ecolab Usa Inc. | Hard surface cleaning compositions comprising phosphinosuccinic acid adducts and methods of use |
| US9752105B2 (en) | 2012-09-13 | 2017-09-05 | Ecolab Usa Inc. | Two step method of cleaning, sanitizing, and rinsing a surface |
| EP2712915A1 (en) | 2012-10-01 | 2014-04-02 | The Procter and Gamble Company | Methods of treating a surface and compositions for use therein |
| EP2904085B1 (en) | 2012-10-04 | 2018-11-07 | Ecolab USA Inc. | Pre-soak technology for laundry and other hard surface cleaning |
| MX361862B (es) | 2012-10-12 | 2018-12-18 | Danisco Us Inc | Composiciones y métodos que comprenden variante de enzima lipolítica. |
| BR112015010104A2 (pt) | 2012-11-05 | 2017-08-22 | Danisco Us Inc | Variante de enzima termolisina, composição e método de limpeza |
| US20150344858A1 (en) | 2012-12-19 | 2015-12-03 | Danisco Us Inc. | Novel mannanase, compositions and methods of use thereof |
| WO2014100100A1 (en) | 2012-12-20 | 2014-06-26 | The Procter & Gamble Company | Detergent composition with silicate coated bleach |
| DK2935575T3 (en) | 2012-12-21 | 2018-07-23 | Danisco Us Inc | ALPHA-amylase variants |
| WO2014099525A1 (en) | 2012-12-21 | 2014-06-26 | Danisco Us Inc. | Paenibacillus curdlanolyticus amylase, and methods of use, thereof |
| EP2767582A1 (en) | 2013-02-19 | 2014-08-20 | The Procter and Gamble Company | Method of laundering a fabric |
| ES2834373T3 (es) | 2013-02-19 | 2021-06-17 | Procter & Gamble | Método para lavado de un tejido |
| EP2767579B1 (en) | 2013-02-19 | 2018-07-18 | The Procter and Gamble Company | Method of laundering a fabric |
| ES2652301T3 (es) | 2013-03-05 | 2018-02-01 | The Procter & Gamble Company | Composiciones tensioactivas mixtas de amida de base azucarada |
| ES2882517T3 (es) | 2013-03-11 | 2021-12-02 | Danisco Us Inc | Variantes combinatorias de alfa-amilasa |
| BR112015023790B1 (pt) | 2013-03-15 | 2021-09-14 | Lubrizol Advanced Materials, Inc | Polímeros de ácido itacônico |
| US9631164B2 (en) | 2013-03-21 | 2017-04-25 | Novozymes A/S | Polypeptides with lipase activity and polynucleotides encoding same |
| CN105073966B (zh) | 2013-03-28 | 2018-03-23 | 宝洁公司 | 包含聚醚胺的清洁组合物 |
| CN103215151B (zh) * | 2013-04-25 | 2015-01-07 | 上海巴方精细化工有限公司 | 矢车菊低刺激护肤香皂 |
| US20160083703A1 (en) | 2013-05-17 | 2016-03-24 | Novozymes A/S | Polypeptides having alpha amylase activity |
| JP6188199B2 (ja) * | 2013-05-20 | 2017-08-30 | ライオン株式会社 | 食器洗い機用洗浄剤 |
| CN103361198B (zh) * | 2013-05-22 | 2015-01-07 | 上海巴方精细化工有限公司 | 川芎消炎复合香皂 |
| MX2015016438A (es) | 2013-05-28 | 2016-03-01 | Procter & Gamble | Composiciones que comprenden tintes fotocromicos para el tratamiento de superficies. |
| US20160108388A1 (en) | 2013-05-29 | 2016-04-21 | Danisco Us Inc. | Novel metalloproteases |
| WO2014194117A2 (en) | 2013-05-29 | 2014-12-04 | Danisco Us Inc. | Novel metalloproteases |
| CN105492603B (zh) | 2013-05-29 | 2022-06-03 | 丹尼斯科美国公司 | 新型金属蛋白酶 |
| EP3004314B1 (en) | 2013-05-29 | 2018-06-20 | Danisco US Inc. | Novel metalloproteases |
| US10378001B2 (en) | 2013-06-27 | 2019-08-13 | Novozymes A/S | Subtilase variants and compositions comprising same |
| JP2016523098A (ja) | 2013-07-04 | 2016-08-08 | ノボザイムス アクティーゼルスカブ | 再付着防止効果を有するポリペプチドおよびそれをコードするポリヌクレオチド |
| MX371497B (es) | 2013-07-19 | 2020-01-31 | Danisco Us Inc | Composiciones y metodos que comprenden una variante de enzima lipolitica. |
| CN117165561A (zh) * | 2013-07-29 | 2023-12-05 | 诺维信公司 | 蛋白酶变体以及对其进行编码的多核苷酸 |
| JP2016533740A (ja) * | 2013-07-29 | 2016-11-04 | ダニスコ・ユーエス・インク | 酵素変異体 |
| CN105593365B (zh) | 2013-09-12 | 2021-08-27 | 丹尼斯科美国公司 | 包含lg12-进化枝蛋白酶变体的组合物和方法 |
| US9834682B2 (en) | 2013-09-18 | 2017-12-05 | Milliken & Company | Laundry care composition comprising carboxylate dye |
| CA2921433A1 (en) | 2013-09-18 | 2015-03-26 | The Procter & Gamble Company | Laundry care composition comprising carboxylate dye |
| CN105555934A (zh) | 2013-09-18 | 2016-05-04 | 宝洁公司 | 包含噻吩偶氮羧化物染料的衣物洗涤护理组合物 |
| WO2015042013A1 (en) | 2013-09-18 | 2015-03-26 | Lubrizol Advanced Materials, Inc. | Stable linear polymers |
| WO2015042086A1 (en) | 2013-09-18 | 2015-03-26 | The Procter & Gamble Company | Laundry care composition comprising carboxylate dye |
| DE102013219467A1 (de) * | 2013-09-26 | 2015-03-26 | Henkel Ag & Co. Kgaa | Stabilisierte Inhibitor-Proteasevarianten |
| EP2857487A1 (en) | 2013-10-07 | 2015-04-08 | WeylChem Switzerland AG | Multi-compartment pouch comprising cleaning compositions, washing process and use for washing and cleaning of textiles and dishes |
| EP2857485A1 (en) | 2013-10-07 | 2015-04-08 | WeylChem Switzerland AG | Multi-compartment pouch comprising alkanolamine-free cleaning compositions, washing process and use for washing and cleaning of textiles and dishes |
| EP2857486A1 (en) | 2013-10-07 | 2015-04-08 | WeylChem Switzerland AG | Multi-compartment pouch comprising cleaning compositions, washing process and use for washing and cleaning of textiles and dishes |
| WO2015057619A1 (en) | 2013-10-15 | 2015-04-23 | Danisco Us Inc. | Clay granule |
| DK3068879T3 (da) | 2013-11-14 | 2020-03-16 | Danisco Us Inc | Stabile enzymer opnået ved glykeringsreduktion |
| US10533165B2 (en) | 2013-12-13 | 2020-01-14 | Danisco Us Inc | Serine proteases of the bacillus gibsonii-clade |
| DK3514230T3 (da) | 2013-12-13 | 2021-11-15 | Danisco Us Inc | Serinproteaser af bacillus-arter |
| AU2014366222B2 (en) | 2013-12-16 | 2018-07-19 | Nutrition & Biosciences USA 4, Inc. | Use of poly alpha-1,3-glucan ethers as viscosity modifiers |
| JP6559139B2 (ja) | 2013-12-18 | 2019-08-14 | イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニーＥ．Ｉ．Ｄｕ Ｐｏｎｔ Ｄｅ Ｎｅｍｏｕｒｓ Ａｎｄ Ｃｏｍｐａｎｙ | カチオン性ポリα−１，３−グルカンエーテル |
| JP6585602B2 (ja) | 2013-12-31 | 2019-10-02 | ダニスコ・ユーエス・インク | タンパク質発現の増大 |
| WO2015112339A1 (en) | 2014-01-22 | 2015-07-30 | The Procter & Gamble Company | Fabric treatment composition |
| EP3097172A1 (en) | 2014-01-22 | 2016-11-30 | The Procter & Gamble Company | Method of treating textile fabrics |
| WO2015112341A1 (en) | 2014-01-22 | 2015-07-30 | The Procter & Gamble Company | Fabric treatment composition |
| WO2015112340A1 (en) | 2014-01-22 | 2015-07-30 | The Procter & Gamble Company | Method of treating textile fabrics |
| CA2841024C (en) | 2014-01-30 | 2017-03-07 | The Procter & Gamble Company | Unit dose article |
| WO2015123323A1 (en) | 2014-02-14 | 2015-08-20 | E. I. Du Pont De Nemours And Company | Poly-alpha-1,3-1,6-glucans for viscosity modification |
| WO2015130669A1 (en) | 2014-02-25 | 2015-09-03 | The Procter & Gamble Company | A process for making renewable surfactant intermediates and surfactants from fats and oils and products thereof |
| WO2015130653A1 (en) | 2014-02-25 | 2015-09-03 | The Procter & Gamble Company | A process for making renewable surfactant intermediates and surfactants from fats and oils and products thereof |
| CN106232791B (zh) | 2014-02-26 | 2019-12-10 | 宝洁公司 | 包含烷氧基化的聚亚烷基亚胺、有机改性的硅氧烷和基于硅氧烷的稀释剂的清洁组合物 |
| EP2915873A1 (en) * | 2014-03-06 | 2015-09-09 | The Procter and Gamble Company | Dishwashing composition |
| ES2734726T3 (es) | 2014-03-11 | 2019-12-11 | Du Pont | Poli alfa-1,2-glucano oxidado como reforzante de detergente |
| KR102350475B1 (ko) | 2014-03-14 | 2022-01-11 | 루브리졸 어드밴스드 머티어리얼스, 인코포레이티드 | 이타콘산 폴리머 및 코폴리머 |
| DK3119884T3 (da) | 2014-03-21 | 2019-10-14 | Danisco Us Inc | Serinproteaser af bacillus-arter |
| CA2941253A1 (en) | 2014-03-27 | 2015-10-01 | Frank Hulskotter | Cleaning compositions containing a polyetheramine |
| EP3122850A1 (en) | 2014-03-27 | 2017-02-01 | The Procter & Gamble Company | Cleaning compositions containing a polyetheramine |
| EP2924105A1 (en) | 2014-03-28 | 2015-09-30 | The Procter and Gamble Company | Water soluble unit dose article |
| EP2924106A1 (en) | 2014-03-28 | 2015-09-30 | The Procter and Gamble Company | Water soluble unit dose article |
| US20170015950A1 (en) | 2014-04-01 | 2017-01-19 | Novozymes A/S | Polypeptides having alpha amylase activity |
| MX2016014383A (es) | 2014-05-06 | 2017-01-20 | Milliken & Co | Composiciones para el cuidado de la ropa. |
| US9365805B2 (en) | 2014-05-15 | 2016-06-14 | Ecolab Usa Inc. | Bio-based pot and pan pre-soak |
| WO2015187757A1 (en) | 2014-06-06 | 2015-12-10 | The Procter & Gamble Company | Detergent composition comprising polyalkyleneimine polymers |
| 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 |
| US9771548B2 (en) | 2014-06-19 | 2017-09-26 | E I Du Pont De Nemours And Company | Compositions containing one or more poly alpha-1,3-glucan ether compounds |
| US9714403B2 (en) | 2014-06-19 | 2017-07-25 | E I Du Pont De Nemours And Company | Compositions containing one or more poly alpha-1,3-glucan ether compounds |
| US10626388B2 (en) | 2014-07-04 | 2020-04-21 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| EP3140399B1 (en) | 2014-07-04 | 2018-03-28 | Novozymes A/S | Subtilase variants and polynucleotides encoding same |
| JP6879902B2 (ja) | 2014-08-01 | 2021-06-02 | エコラボ ユーエスエー インコーポレイティド | 洗浄用布地を用いた手動表面洗浄方法、及び上記洗浄布地を洗う方法 |
| EP2987849A1 (en) | 2014-08-19 | 2016-02-24 | The Procter and Gamble Company | Method of Laundering a Fabric |
| EP2987848A1 (en) | 2014-08-19 | 2016-02-24 | The Procter & Gamble Company | Method of laundering a fabric |
| CN106687572A (zh) | 2014-09-10 | 2017-05-17 | 巴斯夫欧洲公司 | 包封清洁组合物 |
| US9617502B2 (en) | 2014-09-15 | 2017-04-11 | The Procter & Gamble Company | Detergent compositions containing salts of polyetheramines and polymeric acid |
| EP3197988B1 (en) | 2014-09-25 | 2018-08-01 | The Procter & Gamble Company | Cleaning compositions containing a polyetheramine |
| US20160090552A1 (en) | 2014-09-25 | 2016-03-31 | The Procter & Gamble Company | Detergent compositions containing a polyetheramine and an anionic soil release polymer |
| US9388368B2 (en) | 2014-09-26 | 2016-07-12 | The Procter & Gamble Company | Cleaning compositions containing a polyetheramine |
| DK3207129T3 (da) | 2014-10-17 | 2020-02-24 | Danisco Us Inc | Serinproteaser af bacillus-arten |
| EP3172329A1 (en) | 2014-10-24 | 2017-05-31 | Danisco US Inc. | Method for producing alcohol by use of a tripeptidyl peptidase |
| EP3550017B1 (en) | 2014-10-27 | 2021-07-14 | Danisco US Inc. | Serine proteases |
| DK3212662T3 (da) | 2014-10-27 | 2020-07-20 | Danisco Us Inc | Serinproteaser |
| US20180010074A1 (en) | 2014-10-27 | 2018-01-11 | Danisco Us Inc. | Serine proteases of bacillus species |
| DK3212781T3 (da) | 2014-10-27 | 2019-12-16 | Danisco Us Inc | Serinproteaser |
| EP4403631A3 (en) | 2014-10-27 | 2024-10-30 | Danisco US Inc. | Serine proteases |
| EP3256563A1 (en) | 2014-11-17 | 2017-12-20 | The Procter and Gamble Company | Benefit agent delivery compositions |
| US10287562B2 (en) * | 2014-11-20 | 2019-05-14 | Novoszymes A/S | Alicyclobacillus variants and polynucleotides encoding same |
| KR102500121B1 (ko) | 2014-12-16 | 2023-02-20 | 다니스코 유에스 인크. | 향상된 단백질 발현 |
| KR20170093247A (ko) | 2014-12-19 | 2017-08-14 | 다니스코 유에스 인크. | 향상된 단백질 발현 |
| JP6770519B2 (ja) | 2014-12-23 | 2020-10-14 | イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニーＥ．Ｉ．Ｄｕ Ｐｏｎｔ Ｄｅ Ｎｅｍｏｕｒｓ Ａｎｄ Ｃｏｍｐａｎｙ | 酵素により製造されるセルロース |
| WO2016134213A2 (en) | 2015-02-19 | 2016-08-25 | Danisco Us Inc | Enhanced protein expression |
| US20180216090A1 (en) | 2015-03-12 | 2018-08-02 | Danisco Us Inc. | Compositions and methods comprising lg12-clade protease variants |
| WO2016176240A1 (en) | 2015-04-29 | 2016-11-03 | The Procter & Gamble Company | Method of treating a fabric |
| US10513671B2 (en) | 2015-04-29 | 2019-12-24 | The Procter & Gamble Company | Method of treating a fabric |
| PL3088503T3 (pl) | 2015-04-29 | 2018-10-31 | The Procter & Gamble Company | Sposób prania tkanin |
| ES2682178T3 (es) | 2015-04-29 | 2018-09-19 | The Procter & Gamble Company | Composición detergente |
| US20160319227A1 (en) | 2015-04-29 | 2016-11-03 | The Procter & Gamble Company | Method of treating a fabric |
| CN111718806B (zh) | 2015-05-04 | 2022-01-04 | 美利肯公司 | 在洗衣护理组合物中作为上蓝剂的隐色三苯甲烷着色剂 |
| US20180142224A1 (en) | 2015-05-08 | 2018-05-24 | Novozymes A/S | Alpha-amylase variants having improved performance and stability |
| AR105416A1 (es) | 2015-05-08 | 2017-10-04 | Novozymes As | VARIANTES DE a-AMILASA Y POLINUCLEÓTIDOS QUE LAS CODIFICAN |
| AU2016259703B2 (en) | 2015-05-08 | 2021-12-23 | Novozymes A/S | Alpha-amylase variants and polynucleotides encoding same |
| US20180112204A1 (en) | 2015-05-13 | 2018-04-26 | Danisco Us Inc. | AprL-CLADE PROTEASE VARIANTS AND USES THEREOF |
| EP3303536B1 (en) | 2015-06-02 | 2019-04-17 | Unilever PLC | Laundry detergent composition |
| US20180134997A1 (en) | 2015-06-09 | 2018-05-17 | Danisco Us Inc. | Osmotic burst encapsulates |
| BR112017026234B1 (pt) | 2015-06-11 | 2022-10-04 | Unilever Ip Holdings B.V. | Composição de detergente para lavagem de roupas e método de tratamento doméstico de um tecido |
| US11499146B2 (en) | 2015-06-17 | 2022-11-15 | Danisco Us Inc. | Bacillus gibsonii-clade serine proteases |
| EP3313966B1 (en) | 2015-06-26 | 2020-07-29 | Unilever PLC | Laundry detergent composition |
| WO2017046260A1 (en) | 2015-09-17 | 2017-03-23 | Novozymes A/S | Polypeptides having xanthan degrading activity and polynucleotides encoding same |
| AU2016323412B2 (en) | 2015-09-17 | 2021-04-01 | Henkel Ag & Co. Kgaa | Detergent compositions comprising polypeptides having xanthan degrading activity |
| EP3359634A1 (en) * | 2015-10-09 | 2018-08-15 | Novozymes A/S | Laundry method, use of polypeptide and detergent composition |
| US20180320158A1 (en) | 2015-11-05 | 2018-11-08 | Danisco Us Inc. | Paenibacillus and bacillus spp. mannanases |
| JP7364331B2 (ja) | 2015-11-05 | 2023-10-18 | ダニスコ・ユーエス・インク | パエニバチルス（Ｐａｅｎｉｂａｃｉｌｌｕｓ）属種マンナナーゼ |
| WO2017083229A1 (en) | 2015-11-13 | 2017-05-18 | E. I. Du Pont De Nemours And Company | Glucan fiber compositions for use in laundry care and fabric care |
| US10876074B2 (en) | 2015-11-13 | 2020-12-29 | Dupont Industrial Biosciences Usa, Llc | Glucan fiber compositions for use in laundry care and fabric care |
| WO2017083228A1 (en) | 2015-11-13 | 2017-05-18 | E. I. Du Pont De Nemours And Company | Glucan fiber compositions for use in laundry care and fabric care |
| WO2017089093A1 (en) * | 2015-11-25 | 2017-06-01 | Unilever N.V. | A liquid detergent composition |
| DK3387124T3 (da) | 2015-12-09 | 2021-08-23 | William Cuevas | Kombinatoriske alfa-amylasevarianter |
| JP6591278B2 (ja) * | 2015-12-15 | 2019-10-16 | 花王株式会社 | 食器用固体洗浄剤組成物 |
| US20180362946A1 (en) | 2015-12-18 | 2018-12-20 | Danisco Us Inc. | Polypeptides with endoglucanase activity and uses thereof |
| EP4219702A3 (en) | 2015-12-30 | 2023-11-01 | Novozymes A/S | Enzyme variants and polynucleotides encoding the same |
| EP3205393A1 (en) | 2016-02-12 | 2017-08-16 | Basf Se | Process for preparation of microcapsules |
| EP3205392A1 (en) | 2016-02-12 | 2017-08-16 | Basf Se | Microcapsules and process for preparation of microcapsules |
| EP3417039B1 (en) | 2016-02-17 | 2019-07-10 | Unilever PLC | Whitening composition |
| BR112018016674B1 (pt) | 2016-02-17 | 2022-06-07 | Unilever Ip Holdings B.V. | Composição de detergente para lavagem de roupas e método doméstico de tratamento de um tecido |
| EP3419991B1 (en) | 2016-03-04 | 2022-10-26 | Danisco US Inc. | Engineered ribosomal promoters for protein production in microorganisms |
| US20190085273A1 (en) | 2016-03-09 | 2019-03-21 | Basf Se | Encapsulated laundry cleaning composition |
| WO2017162378A1 (en) | 2016-03-21 | 2017-09-28 | Unilever Plc | Laundry detergent composition |
| CN108779416B (zh) | 2016-04-08 | 2021-01-05 | 荷兰联合利华有限公司 | 洗衣洗涤剂组合物 |
| WO2017182295A1 (en) | 2016-04-18 | 2017-10-26 | Basf Se | Liquid cleaning compositions |
| BR112018072530A2 (pt) | 2016-05-03 | 2019-03-26 | Danisco Us Inc | variantes de protease e usos das mesmas |
| BR112018072586A2 (pt) | 2016-05-05 | 2019-02-19 | Danisco Us Inc | variantes de protease e usos das mesmas |
| JP6067168B1 (ja) * | 2016-05-30 | 2017-01-25 | 株式会社ニイタカ | 自動洗浄機用洗浄剤組成物 |
| US11661567B2 (en) | 2016-05-31 | 2023-05-30 | Danisco Us Inc. | Protease variants and uses thereof |
| CN110129298B (zh) * | 2016-06-02 | 2021-08-03 | 天津科技大学 | 一种磷脂酶d突变体及其应用 |
| WO2017219011A1 (en) | 2016-06-17 | 2017-12-21 | Danisco Us Inc | Protease variants and uses thereof |
| EP3275989A1 (en) | 2016-07-26 | 2018-01-31 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| EP3275988B1 (en) | 2016-07-26 | 2020-07-08 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| EP3275985A1 (en) | 2016-07-26 | 2018-01-31 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| EP3275986B1 (en) | 2016-07-26 | 2020-07-08 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| EP3275987A1 (en) | 2016-07-26 | 2018-01-31 | The Procter and Gamble Company | Automatic dishwashing detergent composition |
| AU2017317564B2 (en) | 2016-08-24 | 2021-09-30 | Henkel Ag & Co. Kgaa | Detergent composition comprising GH9 endoglucanase variants I |
| WO2018037061A1 (en) | 2016-08-24 | 2018-03-01 | Novozymes A/S | Xanthan lyase variants and polynucleotides encoding same |
| EP3504330A1 (en) | 2016-08-24 | 2019-07-03 | Novozymes A/S | Gh9 endoglucanase variants and polynucleotides encoding same |
| KR102483218B1 (ko) | 2016-08-24 | 2023-01-02 | 헨켈 아게 운트 코. 카게아아 | 크산탄 리아제 변이체 i을 포함하는 세제 조성물 |
| EP3535319A1 (en) | 2016-11-01 | 2019-09-11 | Milliken & Company | Leuco colorants as bluing agents in laundry care compositions |
| BR112019008706A2 (pt) | 2016-11-01 | 2019-07-16 | Procter & Gamble | colorantes leucos como agentes de azulamento em composições para cuidado na lavagem de roupas |
| CN109890949B (zh) | 2016-11-01 | 2021-10-01 | 宝洁公司 | 衣物洗涤护理组合物中作为上蓝剂的隐色着色剂、其包装、试剂盒和方法 |
| US20180119056A1 (en) | 2016-11-03 | 2018-05-03 | Milliken & Company | Leuco Triphenylmethane Colorants As Bluing Agents in Laundry Care Compositions |
| US20190264138A1 (en) | 2016-11-07 | 2019-08-29 | Danisco Us Inc. | Laundry detergent composition |
| US10577571B2 (en) | 2016-11-08 | 2020-03-03 | Ecolab Usa Inc. | Non-aqueous cleaner for vegetable oil soils |
| CN110023476B (zh) | 2016-12-02 | 2021-07-06 | 宝洁公司 | 包含酶的清洁组合物 |
| EP4001389A1 (en) | 2016-12-02 | 2022-05-25 | The Procter & Gamble Company | Cleaning compositions including enzymes |
| US10550443B2 (en) | 2016-12-02 | 2020-02-04 | The Procter & Gamble Company | Cleaning compositions including enzymes |
| BR112019011999B1 (pt) | 2016-12-15 | 2022-11-08 | Unilever Ip Holdings B.V | Composição de detergente líquida aquosa para lavagem de roupas e método doméstico de tratamento de um tecido |
| WO2018118950A1 (en) | 2016-12-21 | 2018-06-28 | Danisco Us Inc. | Bacillus gibsonii-clade serine proteases |
| CN110312795B (zh) * | 2016-12-21 | 2024-07-23 | 丹尼斯科美国公司 | 蛋白酶变体及其用途 |
| EP3339423A1 (en) | 2016-12-22 | 2018-06-27 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| ES2965826T3 (es) | 2017-02-01 | 2024-04-17 | Procter & Gamble | Composiciones limpiadoras que comprenden variantes de amilasa |
| DK3585910T3 (da) | 2017-02-24 | 2024-06-17 | Danisco Us Inc | Sammensætninger og fremgangsmåder til forøget proteinproduktion i bacillus licheniformis |
| WO2018169750A1 (en) | 2017-03-15 | 2018-09-20 | Danisco Us Inc | Trypsin-like serine proteases and uses thereof |
| US20210095268A1 (en) | 2017-03-31 | 2021-04-01 | Danisco Us Inc | Alpha-amylase combinatorial variants |
| EP3601515A1 (en) | 2017-03-31 | 2020-02-05 | Danisco US Inc. | Delayed release enzyme formulations for bleach-containing detergents |
| 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 |
| EP3645696A1 (en) | 2017-06-30 | 2020-05-06 | Danisco US Inc. | Low-agglomeration, enzyme-containing particles |
| WO2019008035A1 (en) | 2017-07-07 | 2019-01-10 | Unilever Plc | LAUNDRY COMPOSITION FOR LAUNDRY |
| EP3649222B1 (en) | 2017-07-07 | 2024-03-13 | Unilever IP Holdings B.V. | Whitening composition |
| RU2020111041A (ru) | 2017-08-18 | 2021-09-20 | ДАНИСКО ЮЭс ИНК | Варианты альфа-амилаз |
| WO2019040412A1 (en) | 2017-08-23 | 2019-02-28 | Danisco Us Inc | METHODS AND COMPOSITIONS FOR EFFICIENT GENETIC MODIFICATION OF BACILLUS LICHENIFORMIS STRAINS |
| US20210130744A1 (en) | 2017-08-24 | 2021-05-06 | Henkel Ag & Co. Kgaa | Detergent composition comprising xanthan lyase variants ii |
| EP3673056A1 (en) | 2017-08-24 | 2020-07-01 | Henkel AG & Co. KGaA | Detergent compositions comprising gh9 endoglucanase variants ii |
| WO2019038058A1 (en) | 2017-08-24 | 2019-02-28 | Novozymes A/S | GH9 ENDOGLUCANASE VARIANTS AND POLYNUCLEOTIDES ENCODING SUCH VARIANTS |
| EP3673057A1 (en) | 2017-08-24 | 2020-07-01 | Novozymes A/S | Xanthan lyase variants and polynucleotides encoding same |
| JP7218985B2 (ja) | 2017-09-13 | 2023-02-07 | ダニスコ・ユーエス・インク | バチルス属（Ｂａｃｉｌｌｕｓ）におけるタンパク質産生の増加のための改変５’－非翻訳領域（ＵＴＲ）配列 |
| TWI715878B (zh) | 2017-10-12 | 2021-01-11 | 美商美力肯及公司 | 隱色著色劑及組成物 |
| CA3074938A1 (en) | 2017-10-12 | 2019-04-18 | The Procter & Gamble Company | Leuco colorants as bluing agents in laundry care composition |
| WO2019075148A1 (en) | 2017-10-12 | 2019-04-18 | The Procter & Gamble Company | LEUCO-COLORANTS AS AZURING AGENTS IN LAUNDRY CARE COMPOSITIONS |
| CN111194348B (zh) | 2017-10-12 | 2021-12-03 | 宝洁公司 | 衣物洗涤护理组合物中的与作为上蓝剂的第二增白剂组合的隐色着色剂 |
| CN111511842B (zh) | 2017-11-01 | 2022-01-11 | 美利肯公司 | 隐色化合物、着色剂化合物及含有其的组合物 |
| WO2019089898A1 (en) | 2017-11-02 | 2019-05-09 | Danisco Us Inc | Freezing point depressed solid matrix compositions for melt granulation of enzymes |
| EP3717643A1 (en) | 2017-11-29 | 2020-10-07 | Danisco US Inc. | Subtilisin variants having improved stability |
| WO2019105675A1 (en) | 2017-11-30 | 2019-06-06 | Unilever Plc | Detergent composition comprising protease |
| EP3502246A1 (en) | 2017-12-19 | 2019-06-26 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| EP3502244A1 (en) | 2017-12-19 | 2019-06-26 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| EP3502245A1 (en) | 2017-12-19 | 2019-06-26 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| EP3502227B1 (en) | 2017-12-19 | 2024-09-04 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| DE102017223281A1 (de) * | 2017-12-19 | 2019-06-19 | Henkel Ag & Co. Kgaa | Reinigugnsmittel enthaltend durch Betain stabilisierte Amylase |
| US12134791B2 (en) | 2017-12-21 | 2024-11-05 | Danisco Us Inc. | Enzyme-containing, hot-melt granules comprising a thermotolerant desiccant |
| JP6967447B2 (ja) * | 2017-12-27 | 2021-11-17 | 小林製薬株式会社 | 錠剤状義歯洗浄剤 |
| US11414643B2 (en) | 2018-01-03 | 2022-08-16 | Danisco Us Inc. | Mutant and genetically modified Bacillus cells and methods thereof for increased protein production |
| KR20200086739A (ko) | 2018-01-26 | 2020-07-17 | 더 프록터 앤드 갬블 캄파니 | 효소를 포함하는 수용성 단위 용량 물품 |
| WO2019156670A1 (en) | 2018-02-08 | 2019-08-15 | Danisco Us Inc. | Thermally-resistant wax matrix particles for enzyme encapsulation |
| EP3755793A1 (en) | 2018-02-23 | 2020-12-30 | Henkel AG & Co. KGaA | Detergent composition comprising xanthan lyase and endoglucanase variants |
| US20190264139A1 (en) | 2018-02-28 | 2019-08-29 | The Procter & Gamble Company | Cleaning compositions |
| US20190264140A1 (en) | 2018-02-28 | 2019-08-29 | The Procter & Gamble Company | Methods of cleaning |
| WO2019180111A1 (en) * | 2018-03-23 | 2019-09-26 | Novozymes A/S | Subtilase variants and compositions comprising same |
| CN112119147B (zh) | 2018-05-17 | 2023-09-29 | 联合利华知识产权控股有限公司 | 清洁组合物 |
| US10913919B2 (en) | 2018-06-19 | 2021-02-09 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| EP3810769A1 (en) | 2018-06-19 | 2021-04-28 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| US20210214703A1 (en) | 2018-06-19 | 2021-07-15 | Danisco Us Inc | Subtilisin variants |
| US20210363470A1 (en) | 2018-06-19 | 2021-11-25 | Danisco Us Inc | Subtilisin variants |
| US20200032178A1 (en) | 2018-07-27 | 2020-01-30 | The Procter & Gamble Company | Water-soluble unit dose articles comprising water-soluble fibrous structures and particles |
| CA3108284A1 (en) | 2018-07-31 | 2020-02-06 | Danisco Us Inc | Variant alpha-amylases having amino acid substitutions that lower the pka of the general acid |
| US20240279627A1 (en) | 2018-08-30 | 2024-08-22 | Danisco Us Inc | Compositions comprising a lipolytic enzyme variant and methods of use thereof |
| WO2020047215A1 (en) | 2018-08-30 | 2020-03-05 | Danisco Us Inc | Enzyme-containing granules |
| EP3853330B1 (en) | 2018-09-17 | 2023-06-07 | Unilever Global Ip Limited | Detergent composition |
| JP2022503923A (ja) | 2018-09-27 | 2022-01-12 | ダニスコ・ユーエス・インク | 医療用器具を洗浄するための組成物 |
| CN113166745A (zh) | 2018-10-12 | 2021-07-23 | 丹尼斯科美国公司 | 在螯合剂存在下具有可增强稳定性的突变的α-淀粉酶 |
| WO2020104159A1 (en) | 2018-11-20 | 2020-05-28 | Unilever Plc | Detergent composition |
| CN113056548B (zh) | 2018-11-20 | 2023-05-02 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| CN113056550B (zh) | 2018-11-20 | 2022-10-28 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| CN113056549B (zh) | 2018-11-20 | 2023-03-10 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| WO2020104156A1 (en) | 2018-11-20 | 2020-05-28 | Unilever Plc | Detergent composition |
| US20230028935A1 (en) | 2018-11-28 | 2023-01-26 | Danisco Us Inc | Subtilisin variants having improved stability |
| CN113330101A (zh) | 2018-12-21 | 2021-08-31 | 诺维信公司 | 包含金属蛋白酶的洗涤剂袋 |
| EP3702452A1 (en) * | 2019-03-01 | 2020-09-02 | Novozymes A/S | Detergent compositions comprising two proteases |
| CN113439116B (zh) | 2019-03-14 | 2023-11-28 | 宝洁公司 | 包含酶的清洁组合物 |
| EP3938502A1 (en) | 2019-03-14 | 2022-01-19 | The Procter & Gamble Company | Cleaning compositions comprising enzymes |
| WO2020186052A1 (en) | 2019-03-14 | 2020-09-17 | The Procter & Gamble Company | Method for treating cotton |
| BR112021018731A2 (pt) | 2019-03-21 | 2021-12-21 | Novozymes As | Variantes de alfa-amilase e polinucleotídeos codificando as mesmas |
| US20220195337A1 (en) | 2019-05-16 | 2022-06-23 | Conopco, Inc., D/B/A Unilever | Laundry composition |
| US11261404B2 (en) | 2019-05-20 | 2022-03-01 | Ecolab Usa Inc. | Surfactant package for high foaming detergents with low level of medium to long chain linear alcohols |
| EP3741283A1 (en) | 2019-05-22 | 2020-11-25 | The Procter & Gamble Company | Automatic dishwashing method |
| WO2020243738A1 (en) | 2019-05-24 | 2020-12-03 | The Procter & Gamble Company | Automatic dishwashing detergent composition |
| EP3976776A1 (en) | 2019-05-24 | 2022-04-06 | Danisco US Inc. | Subtilisin variants and methods of use |
| EP3980517A1 (en) | 2019-06-06 | 2022-04-13 | Danisco US Inc. | Methods and compositions for cleaning |
| CN118834855A (zh) | 2019-06-24 | 2024-10-25 | 宝洁公司 | 包含淀粉酶变体的清洁组合物 |
| WO2020260006A1 (en) | 2019-06-28 | 2020-12-30 | Unilever Plc | Detergent compositions |
| US20220372397A1 (en) | 2019-06-28 | 2022-11-24 | Conopco, Inc., D/B/A Unilever | Detergent composition |
| WO2020259949A1 (en) | 2019-06-28 | 2020-12-30 | Unilever Plc | Detergent composition |
| US20220364022A1 (en) | 2019-06-28 | 2022-11-17 | Conopco, Inc., D/B/A Unilever | Detergent composition |
| CN113891930A (zh) | 2019-06-28 | 2022-01-04 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| BR112021025399A2 (pt) | 2019-06-28 | 2022-04-26 | Unilever Ip Holdings B V | Composição detergente e método doméstico para tratar um tecido |
| EP3770241A1 (de) * | 2019-07-22 | 2021-01-27 | Henkel AG & Co. KGaA | Reinigungsmittel mit protease zur automatischen dosierung |
| US11873465B2 (en) | 2019-08-14 | 2024-01-16 | Ecolab Usa Inc. | Methods of cleaning and soil release of highly oil absorbing substrates employing optimized extended chain nonionic surfactants |
| CN114364776A (zh) | 2019-09-02 | 2022-04-15 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| US20220372399A1 (en) | 2019-09-19 | 2022-11-24 | Conopco, Inc., D/B/A Unilever | Detergent compositions |
| AR120142A1 (es) | 2019-10-07 | 2022-02-02 | Unilever Nv | Composición detergente |
| BR112022007697A2 (pt) | 2019-10-24 | 2022-07-12 | Danisco Us Inc | Alfa-amilase variante que forma maltopentaose/maltohexaose |
| US11492571B2 (en) | 2019-10-24 | 2022-11-08 | The Procter & Gamble Company | Automatic dishwashing detergent composition comprising a protease |
| US20210122998A1 (en) | 2019-10-24 | 2021-04-29 | The Procter & Gamble Company | Automatic dishwashing detergent composition comprising an amylase |
| KR102316445B1 (ko) * | 2019-11-29 | 2021-10-26 | 씨제이제일제당 주식회사 | 신규 세린 프로테아제 변이체 |
| EP3835396A1 (en) | 2019-12-09 | 2021-06-16 | The Procter & Gamble Company | A detergent composition comprising a polymer |
| EP4090727A1 (en) | 2020-01-13 | 2022-11-23 | Danisco US Inc. | Compositions comprising a lipolytic enzyme variant and methods of use thereof |
| KR20220127844A (ko) | 2020-01-15 | 2022-09-20 | 다니스코 유에스 인크. | 바실러스 리체니포르미스에서 단백질 생산을 향상시키기 위한 조성물 및 방법 |
| EP4097206B1 (en) | 2020-01-29 | 2023-08-09 | Unilever IP Holdings B.V. | Laundry detergent product |
| EP3862412A1 (en) | 2020-02-04 | 2021-08-11 | The Procter & Gamble Company | Detergent composition |
| CA3173147A1 (en) | 2020-06-05 | 2021-12-09 | Phillip Kyle Vinson | Detergent compositions containing a branched surfactant |
| CN115698246A (zh) | 2020-06-08 | 2023-02-03 | 联合利华知识产权控股有限公司 | 提高蛋白酶活性的方法 |
| WO2022010893A1 (en) | 2020-07-06 | 2022-01-13 | Ecolab Usa Inc. | Foaming mixed alcohol/water compositions comprising a combination of alkyl siloxane and a hydrotrope/solubilizer |
| WO2022010911A1 (en) | 2020-07-06 | 2022-01-13 | Ecolab Usa Inc. | Foaming mixed alcohol/water compositions comprising a structured alkoxylated siloxane |
| US20220002636A1 (en) | 2020-07-06 | 2022-01-06 | Ecolab Usa Inc. | Peg-modified castor oil based compositions for microemulsifying and removing multiple oily soils |
| WO2022031310A1 (en) | 2020-08-04 | 2022-02-10 | The Procter & Gamble Company | Automatic dishwashing method |
| CA3187735A1 (en) | 2020-08-04 | 2022-02-10 | Nina Elizabeth GRAY | Automatic dishwashing method and pack |
| WO2022031309A1 (en) | 2020-08-04 | 2022-02-10 | The Procter & Gamble Company | Automatic dishwashing method |
| WO2022031311A1 (en) | 2020-08-04 | 2022-02-10 | The Procter & Gamble Company | Automatic dishwashing method |
| CN116323935A (zh) | 2020-08-27 | 2023-06-23 | 丹尼斯科美国公司 | 用于清洁的酶和酶组合物 |
| US20230287302A1 (en) | 2020-08-28 | 2023-09-14 | Conopco, Inc., D/B/A Unilever | Detergent composition |
| BR112023001773A2 (pt) | 2020-08-28 | 2023-03-28 | Unilever Ip Holdings B V | Composição detergente e método |
| CN116157496A (zh) | 2020-08-28 | 2023-05-23 | 联合利华知识产权控股有限公司 | 表面活性剂和洗涤剂组合物 |
| EP4204531B1 (en) | 2020-08-28 | 2024-06-26 | Unilever IP Holdings B.V. | Detergent composition |
| WO2022043042A1 (en) | 2020-08-28 | 2022-03-03 | Unilever Ip Holdings B.V. | Detergent composition |
| WO2022074037A2 (en) | 2020-10-07 | 2022-04-14 | Novozymes A/S | Alpha-amylase variants |
| WO2022093189A1 (en) | 2020-10-27 | 2022-05-05 | Milliken & Company | Compositions comprising leuco compounds and colorants |
| MX2023004260A (es) | 2020-10-29 | 2023-04-25 | Procter & Gamble | Composiciones de limpieza que contienen enzimas alginato liasas. |
| EP4001388A1 (en) | 2020-11-17 | 2022-05-25 | The Procter & Gamble Company | Automatic dishwashing method with amphiphilic graft polymer in the rinse |
| WO2022108766A1 (en) | 2020-11-17 | 2022-05-27 | The Procter & Gamble Company | Automatic dishwashing composition comprising amphiphilic graft polymer |
| CA3197110A1 (en) | 2020-11-17 | 2022-05-27 | Patrick Firmin August Delplancke | Automatic dishwashing method with alkaline rinse |
| EP4006131A1 (en) | 2020-11-30 | 2022-06-01 | The Procter & Gamble Company | Method of laundering fabric |
| US20240010950A1 (en) | 2020-12-07 | 2024-01-11 | Conopco Inc., D/B/A Unilever | Detergent compositions |
| AU2021394636A1 (en) | 2020-12-07 | 2023-06-08 | Unilever Global Ip Limited | Detergent 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 |
| MX2023006625A (es) | 2020-12-23 | 2023-07-04 | Procter & Gamble | Poliaminas alcoxiladas anfifilicas y sus usos. |
| CN112662652A (zh) * | 2021-01-20 | 2021-04-16 | 天津科技大学 | 一种胶原降解活力降低的碱性蛋白酶突变体 |
| CN116997642A (zh) | 2021-01-29 | 2023-11-03 | 丹尼斯科美国公司 | 清洁组合物及其相关的方法 |
| EP4039806A1 (en) | 2021-02-04 | 2022-08-10 | Henkel AG & Co. KGaA | Detergent composition comprising xanthan lyase and endoglucanase variants with im-proved stability |
| CN116829709A (zh) | 2021-02-12 | 2023-09-29 | 诺维信公司 | α-淀粉酶变体 |
| CN116615536A (zh) | 2021-03-15 | 2023-08-18 | 宝洁公司 | 含有多肽变体的清洁组合物 |
| CN117157382A (zh) | 2021-05-05 | 2023-12-01 | 宝洁公司 | 制备清洁组合物及检测污垢的方法 |
| EP4086330A1 (en) | 2021-05-06 | 2022-11-09 | The Procter & Gamble Company | Surface treatment |
| CN117677689A (zh) | 2021-05-18 | 2024-03-08 | 诺力昂化学品国际有限公司 | 在清洁应用中的聚酯聚季铵盐 |
| WO2022243533A1 (en) | 2021-05-20 | 2022-11-24 | Nouryon Chemicals International B.V. | Manufactured polymers having altered oligosaccharide or polysaccharide functionality or narrowed oligosaccharide distribution, processes for preparing them, compositions containing them, and methods of using them |
| EP4108767A1 (en) | 2021-06-22 | 2022-12-28 | The Procter & Gamble Company | Cleaning or treatment compositions containing nuclease enzymes |
| WO2022268885A1 (en) | 2021-06-23 | 2022-12-29 | Novozymes A/S | Alpha-amylase polypeptides |
| CN117940546A (zh) | 2021-06-30 | 2024-04-26 | 诺力昂化学品国际有限公司 | 螯合物-两性表面活性剂液体浓缩物及其在清洁应用中的用途 |
| US20240294888A1 (en) | 2021-06-30 | 2024-09-05 | Danisco Us Inc. | Variant enzymes and uses thereof |
| EP4123007A1 (en) | 2021-07-19 | 2023-01-25 | The Procter & Gamble Company | Fabric treatment using bacterial spores |
| EP4123006A1 (en) | 2021-07-19 | 2023-01-25 | The Procter & Gamble Company | Composition comprising spores and pro-perfume materials |
| WO2023017794A1 (ja) | 2021-08-10 | 2023-02-16 | 株式会社日本触媒 | ポリアルキレンオキシド含有化合物 |
| EP4388077A1 (en) | 2021-08-20 | 2024-06-26 | Danisco US Inc. | Polynucleotides encoding novel nucleases, compositions thereof and methods thereof for eliminating dna from protein preparations |
| CN117916354A (zh) | 2021-09-03 | 2024-04-19 | 丹尼斯科美国公司 | 用于清洁的衣物洗涤组合物 |
| WO2023039270A2 (en) | 2021-09-13 | 2023-03-16 | Danisco Us Inc. | Bioactive-containing granules |
| CN117957300A (zh) | 2021-09-20 | 2024-04-30 | 联合利华知识产权控股有限公司 | 洗涤剂组合物 |
| WO2023057367A1 (en) | 2021-10-08 | 2023-04-13 | Unilever Ip Holdings B.V. | Laundry composition |
| EP4416255A1 (en) | 2021-10-14 | 2024-08-21 | The Procter & Gamble Company | A fabric and home care product comprising cationic soil release polymer and lipase enzyme |
| EP4194536A1 (en) | 2021-12-08 | 2023-06-14 | The Procter & Gamble Company | Laundry treatment cartridge |
| EP4194537A1 (en) | 2021-12-08 | 2023-06-14 | The Procter & Gamble Company | Laundry treatment cartridge |
| WO2023114939A2 (en) | 2021-12-16 | 2023-06-22 | Danisco Us Inc. | Subtilisin variants and methods of use |
| WO2023114932A2 (en) | 2021-12-16 | 2023-06-22 | Danisco Us Inc. | Subtilisin variants and methods of use |
| CA3240638A1 (en) | 2021-12-16 | 2023-06-22 | Michelle Jackson | Fabric and home care composition comprising a protease |
| CN118369413A (zh) | 2021-12-16 | 2024-07-19 | 宝洁公司 | 包含淀粉酶的家庭护理组合物 |
| WO2023114793A1 (en) | 2021-12-16 | 2023-06-22 | The Procter & Gamble Company | Home care composition |
| CN118679251A (zh) | 2021-12-16 | 2024-09-20 | 丹尼斯科美国公司 | 枯草杆菌蛋白酶变体和使用方法 |
| CN118647716A (zh) | 2021-12-16 | 2024-09-13 | 丹尼斯科美国公司 | 成麦芽五糖/麦芽六糖变体α-淀粉酶 |
| WO2023114795A1 (en) | 2021-12-16 | 2023-06-22 | The Procter & Gamble Company | Automatic dishwashing composition comprising a protease |
| WO2023168234A1 (en) | 2022-03-01 | 2023-09-07 | Danisco Us Inc. | Enzymes and enzyme compositions for cleaning |
| EP4273209A1 (en) | 2022-05-04 | 2023-11-08 | The Procter & Gamble Company | Machine-cleaning compositions containing enzymes |
| EP4273210A1 (en) | 2022-05-04 | 2023-11-08 | The Procter & Gamble Company | Detergent compositions containing enzymes |
| EP4279571A1 (en) | 2022-05-19 | 2023-11-22 | The Procter & Gamble Company | Laundry composition comprising spores |
| WO2023227335A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Liquid composition comprising linear alkyl benzene sulphonate, methyl ester ethoxylate and alkoxylated zwitterionic polyamine polymer |
| WO2023227421A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Laundry liquid composition comprising a surfactant, an alkoxylated zwitterionic polyamine polymer, and a fragrance |
| WO2023227375A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Laundry liquid composition comprising a surfactant, an aminocarboxylate, an organic acid and a fragrance |
| WO2023227331A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Composition comprising a specific methyl ester ethoxylate surfactant and a lipase |
| WO2023227356A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Composition containing enzyme |
| WO2023227332A1 (en) | 2022-05-27 | 2023-11-30 | Unilever Ip Holdings B.V. | Laundry liquid composition comprising a surfactant, an alkoxylated zwitterionic polyamine polymer and a protease |
| WO2023233026A1 (en) | 2022-06-03 | 2023-12-07 | Unilever Ip Holdings B.V. | Laundry detergent product |
| WO2023250301A1 (en) | 2022-06-21 | 2023-12-28 | Danisco Us Inc. | Methods and compositions for cleaning comprising a polypeptide having thermolysin activity |
| US20240026248A1 (en) | 2022-07-20 | 2024-01-25 | Ecolab Usa Inc. | Novel nonionic extended surfactants, compositions and methods of use thereof |
| EP4321604A1 (en) | 2022-08-08 | 2024-02-14 | The Procter & Gamble Company | A fabric and home care composition comprising surfactant and a polyester |
| WO2024050343A1 (en) | 2022-09-02 | 2024-03-07 | Danisco Us Inc. | Subtilisin variants and methods related thereto |
| WO2024050339A1 (en) | 2022-09-02 | 2024-03-07 | Danisco Us Inc. | Mannanase variants and methods of use |
| WO2024050346A1 (en) | 2022-09-02 | 2024-03-07 | Danisco Us Inc. | Detergent compositions and methods related thereto |
| WO2024056333A1 (en) | 2022-09-13 | 2024-03-21 | Unilever Ip Holdings B.V. | Washing machine and washing method |
| WO2024056334A1 (en) | 2022-09-13 | 2024-03-21 | Unilever Ip Holdings B.V. | Washing machine and washing method |
| WO2024056278A1 (en) | 2022-09-13 | 2024-03-21 | Unilever Ip Holdings B.V. | Washing machine and washing method |
| WO2024056331A1 (en) | 2022-09-13 | 2024-03-21 | Unilever Ip Holdings B.V. | Washing machine and washing method |
| EP4349948A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4349944A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4349942A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4349947A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4349943A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4349946A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Unit dose fabric treatment product |
| EP4349945A1 (en) | 2022-10-05 | 2024-04-10 | Unilever IP Holdings B.V. | Laundry liquid composition |
| EP4361239A1 (en) | 2022-10-25 | 2024-05-01 | Unilever IP Holdings B.V. | Laundry liquid composition |
| WO2024088706A1 (en) | 2022-10-25 | 2024-05-02 | Unilever Ip Holdings B.V. | Composition |
| WO2024088716A1 (en) | 2022-10-25 | 2024-05-02 | Unilever Ip Holdings B.V. | Composition |
| WO2024094778A1 (en) | 2022-11-04 | 2024-05-10 | Clariant International Ltd | Polyesters |
| WO2024094803A1 (en) | 2022-11-04 | 2024-05-10 | The Procter & Gamble Company | Fabric and home care composition |
| WO2024094800A1 (en) | 2022-11-04 | 2024-05-10 | The Procter & Gamble Company | Fabric and home care composition |
| WO2024102698A1 (en) | 2022-11-09 | 2024-05-16 | Danisco Us Inc. | Subtilisin variants and methods of use |
| WO2024115106A1 (en) | 2022-11-29 | 2024-06-06 | Unilever Ip Holdings B.V. | Composition |
| WO2024119298A1 (en) | 2022-12-05 | 2024-06-13 | The Procter & Gamble Company | Fabric and home care composition comprising a polyalkylenecarbonate compound |
| WO2024129520A1 (en) | 2022-12-12 | 2024-06-20 | The Procter & Gamble Company | Fabric and home care composition |
| EP4386074A1 (en) | 2022-12-16 | 2024-06-19 | The Procter & Gamble Company | Fabric and home care composition |
| EP4388967A1 (en) | 2022-12-19 | 2024-06-26 | The Procter & Gamble Company | Dishwashing method |
| WO2024131880A2 (en) | 2022-12-23 | 2024-06-27 | Novozymes A/S | Detergent composition comprising catalase and amylase |
| US20240263162A1 (en) | 2023-02-01 | 2024-08-08 | The Procter & Gamble Company | Detergent compositions containing enzymes |
| WO2024163584A1 (en) | 2023-02-01 | 2024-08-08 | Danisco Us Inc. | Subtilisin variants and methods of use |
| WO2024186819A1 (en) | 2023-03-06 | 2024-09-12 | Danisco Us Inc. | Subtilisin variants and methods of use |
| WO2024191711A1 (en) | 2023-03-16 | 2024-09-19 | Nutrition & Biosciences USA 4, Inc. | Brevibacillus fermentate extracts for cleaning and malodor control and use thereof |
| WO2024194190A1 (en) | 2023-03-17 | 2024-09-26 | Unilever Ip Holdings B.V. | Composition |
| WO2024193937A1 (en) | 2023-03-17 | 2024-09-26 | Unilever Ip Holdings B.V. | Machine dishwash filter cleaner |
| WO2024194098A1 (en) | 2023-03-21 | 2024-09-26 | Unilever Ip Holdings B.V. | Detergent unit dose |
| WO2024213430A1 (en) | 2023-04-11 | 2024-10-17 | Unilever Ip Holdings B.V. | Composition |
| WO2024213443A1 (en) | 2023-04-11 | 2024-10-17 | Unilever Ip Holdings B.V. | Composition |
| WO2024213438A1 (en) | 2023-04-11 | 2024-10-17 | Unilever Ip Holdings B.V. | Composition |
| WO2024213376A1 (en) | 2023-04-11 | 2024-10-17 | Unilever Ip Holdings B.V. | Composition |
| WO2024213428A1 (en) | 2023-04-11 | 2024-10-17 | Unilever Ip Holdings B.V. | Composition |
| WO2024223218A1 (en) | 2023-04-25 | 2024-10-31 | Unilever Ip Holdings B.V. | Composition |
| EP4458932A1 (en) | 2023-05-04 | 2024-11-06 | The Procter & Gamble Company | A fabric and home care composition |
| EP4458933A1 (en) | 2023-05-05 | 2024-11-06 | The Procter & Gamble Company | A fabric and home care composition comprising a propoxylated polyol |
Family Cites Families (94)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1611703B1 (de) * | 1967-10-11 | 1972-03-09 | Windmoeller & Hoelscher | Vorrichtung zum Abteilen einer bestimmten Stueckzahl innerhalb einer am Ende einer Sack- oder Beutelmaschine gebildeten kontinuierlichen Folge von Saecken oder Beuteln |
| US4028263A (en) * | 1973-08-24 | 1977-06-07 | Colgate-Palmolive Company | Bleaching and brightening detergent composition |
| NL7904044A (nl) | 1978-05-31 | 1979-12-04 | Unilever Nv | Werkwijze ter bereiding van homogene, vloeibare compo- sities. |
| US4261868A (en) | 1979-08-08 | 1981-04-14 | Lever Brothers Company | Stabilized enzymatic liquid detergent composition containing a polyalkanolamine and a boron compound |
| US4404128A (en) | 1981-05-29 | 1983-09-13 | The Procter & Gamble Company | Enzyme detergent composition |
| GR76237B (zh) | 1981-08-08 | 1984-08-04 | Procter & Gamble | |
| JPS58144105A (ja) | 1982-02-12 | 1983-08-27 | Kurabo Ind Ltd | スケ−ル除去獣毛繊維の製法 |
| GB8310080D0 (en) | 1983-04-14 | 1983-05-18 | Interox Chemicals Ltd | Bleach composition |
| US4760025A (en) | 1984-05-29 | 1988-07-26 | Genencor, Inc. | Modified enzymes and methods for making same |
| US5264366A (en) | 1984-05-29 | 1993-11-23 | Genencor, Inc. | Protease deficient bacillus |
| US5182204A (en) | 1984-05-29 | 1993-01-26 | Genencor International, Inc. | Non-human carbonyl hydrolase mutants, vectors encoding same and hosts transformed with said vectors |
| US5204015A (en) | 1984-05-29 | 1993-04-20 | Genencor International, Inc. | Subtilisin mutants |
| JPS6141481U (ja) * | 1984-08-10 | 1986-03-17 | 三菱電機株式会社 | シ−ム溶接装置 |
| US4797362A (en) * | 1985-06-06 | 1989-01-10 | Lion Corporation | Alkaline proteases and microorganisms producing same |
| US4908773A (en) * | 1987-04-06 | 1990-03-13 | Genex Corporation | Computer designed stabilized proteins and method for producing same |
| US4728455A (en) | 1986-03-07 | 1988-03-01 | Lever Brothers Company | Detergent bleach compositions, bleaching agents and bleach activators |
| ES2068181T3 (es) | 1986-04-30 | 1995-04-16 | Genencor Int | Mutantes de la carbonil hidrolasa no humana, secuencias de adn y vectores que codifican las mismas y huespedes transformados con dichos vectores. |
| GB8629837D0 (en) | 1986-12-13 | 1987-01-21 | Interox Chemicals Ltd | Bleach activation |
| US4853871A (en) * | 1987-04-06 | 1989-08-01 | Genex Corporation | Computer-based method for designing stablized proteins |
| US5314692A (en) | 1987-08-24 | 1994-05-24 | Cultor Ltd. | Enzyme premix for feed and method |
| WO1989004552A1 (en) | 1987-10-30 | 1989-05-18 | Lsi Logic Corporation | Method and means of fabricating a semiconductor device package |
| DK6488D0 (da) * | 1988-01-07 | 1988-01-07 | Novo Industri As | Enzymer |
| CN1056187C (zh) * | 1988-02-11 | 2000-09-06 | 金克克国际有限公司 | 新的蛋白水解酶及其在洗涤剂中的应用 |
| US5543302A (en) * | 1988-05-27 | 1996-08-06 | Solvay Enzymes, Inc. | Proteases of altered stability to autolytic degradation |
| DE3841152A1 (de) | 1988-12-07 | 1990-06-13 | Hoechst Ag | Verwendung von bakterienlysierendem enzymprodukt als zusatzstoff zur verbesserung der futterverwertung in der tierproduktion |
| GB8908416D0 (en) | 1989-04-13 | 1989-06-01 | Unilever Plc | Bleach activation |
| WO1990014420A1 (en) * | 1989-05-17 | 1990-11-29 | Amgen Inc. | Multiply mutated subtilisins |
| US5665587A (en) * | 1989-06-26 | 1997-09-09 | Novo Nordisk A/S | Modified subtilisins and detergent compositions containing same |
| DK316989D0 (da) * | 1989-06-26 | 1989-06-26 | Novo Nordisk As | Enzymer |
| CA2034486A1 (en) * | 1989-06-26 | 1990-12-27 | Eric Casteleijn | Enzymatic detergent compositions |
| GB9003741D0 (en) | 1990-02-19 | 1990-04-18 | Unilever Plc | Bleach activation |
| ES2100924T3 (es) | 1990-05-21 | 1997-07-01 | Unilever Nv | Activacion de blanqueador. |
| ES2110488T3 (es) * | 1990-12-21 | 1998-02-16 | Novo Nordisk As | Mutantes enzimaticos con un bajo grado de variacion de la carga molecular en un intervalo de ph. |
| US5769630A (en) * | 1991-02-25 | 1998-06-23 | Louisiana State University, | Subperiosteal bone anchor |
| GB9108136D0 (en) | 1991-04-17 | 1991-06-05 | Unilever Plc | Concentrated detergent powder compositions |
| US5340735A (en) * | 1991-05-29 | 1994-08-23 | Cognis, Inc. | Bacillus lentus alkaline protease variants with increased stability |
| US5274147A (en) | 1991-07-11 | 1993-12-28 | Lever Brothers Company, Division Of Conopco, Inc. | Process for preparing manganese complexes |
| EP0995801A1 (de) * | 1991-07-27 | 2000-04-26 | Genencor International GmbH | Verfahren zur Verbesserung des Stabilität von Enzymen und stabilisierte Enzyme |
| GB9118242D0 (en) | 1991-08-23 | 1991-10-09 | Unilever Plc | Machine dishwashing composition |
| GB9124581D0 (en) | 1991-11-20 | 1992-01-08 | Unilever Plc | Bleach catalyst composition,manufacture and use thereof in detergent and/or bleach compositions |
| US5153161A (en) | 1991-11-26 | 1992-10-06 | Lever Brothers Company, Division Of Conopco, Inc. | Synthesis of manganese oxidation catalyst |
| US5194416A (en) | 1991-11-26 | 1993-03-16 | Lever Brothers Company, Division Of Conopco, Inc. | Manganese catalyst for activating hydrogen peroxide bleaching |
| EP0544490A1 (en) | 1991-11-26 | 1993-06-02 | Unilever Plc | Detergent bleach compositions |
| GB9127060D0 (en) | 1991-12-20 | 1992-02-19 | Unilever Plc | Bleach activation |
| CA2085642A1 (en) | 1991-12-20 | 1993-06-21 | Ronald Hage | Bleach activation |
| US5316935A (en) * | 1992-04-06 | 1994-05-31 | California Institute Of Technology | Subtilisin variants suitable for hydrolysis and synthesis in organic media |
| CA2097000A1 (en) * | 1992-05-27 | 1993-11-28 | Norio Moriya | Alkaline lipase, method for producing the same, microorganism producing the same and detergent composition containing alkaline lipase |
| DE4218448A1 (de) | 1992-06-04 | 1993-12-09 | Solvay Enzymes Gmbh & Co Kg | Alkalische Proteasen aus Bacillus pumilus |
| US5256779A (en) | 1992-06-18 | 1993-10-26 | Lever Brothers Company, Division Of Conopco, Inc. | Synthesis of manganese oxidation catalyst |
| US5284944A (en) | 1992-06-30 | 1994-02-08 | Lever Brothers Company, Division Of Conopco, Inc. | Improved synthesis of 1,4,7-triazacyclononane |
| AU682314B2 (en) * | 1992-07-17 | 1997-10-02 | Genencor International, Inc. | High alkaline serine proteases |
| US5280117A (en) | 1992-09-09 | 1994-01-18 | Lever Brothers Company, A Division Of Conopco, Inc. | Process for the preparation of manganese bleach catalyst |
| US5567601A (en) * | 1993-06-01 | 1996-10-22 | University Of Maryland | Subtilisin mutants lacking a primary calcium binding site |
| AU7524994A (en) * | 1993-08-12 | 1995-03-14 | University Of Maryland | Thermostable alkaline metalloprotease produced by a hyphomonas, and preparation thereof |
| US6436690B1 (en) | 1993-09-15 | 2002-08-20 | The Procter & Gamble Company | BPN′ variants having decreased adsorption and increased hydrolysis wherein one or more loop regions are substituted |
| MA23346A1 (fr) * | 1993-10-14 | 1995-04-01 | Genencor Int | Variantes de la subtilisine |
| CA2173106C (en) * | 1993-10-14 | 2000-02-15 | Michael Eugene Burns | Bleaching compositions comprising protease enzymes |
| CA2173105C (en) * | 1993-10-14 | 2003-05-27 | Andre Baeck | Protease-containing cleaning compositions |
| EP0724391B1 (en) * | 1993-10-23 | 1999-05-06 | Imperial Tobacco Limited | Improvements in or relating to smoking articles |
| ES2364776T3 (es) * | 1994-02-24 | 2011-09-14 | HENKEL AG & CO. KGAA | Enzimas mejoradas y detergentes que las contienen. |
| ATE390487T1 (de) * | 1994-02-24 | 2008-04-15 | Henkel Kgaa | Verbesserte enzyme und diese enthaltene detergentien |
| US5691295A (en) * | 1995-01-17 | 1997-11-25 | Cognis Gesellschaft Fuer Biotechnologie Mbh | Detergent compositions |
| US5824531A (en) * | 1994-03-29 | 1998-10-20 | Novid Nordisk | Alkaline bacilus amylase |
| DE69504489T2 (de) * | 1994-04-07 | 1999-05-20 | The Procter & Gamble Co., Cincinnati, Ohio | Bleichmittel enthaltend metall haltige bleichkatalysatoren und antioxidantien |
| US6599730B1 (en) * | 1994-05-02 | 2003-07-29 | Procter & Gamble Company | Subtilisin 309 variants having decreased adsorption and increased hydrolysis |
| GB9409336D0 (en) | 1994-05-10 | 1994-06-29 | Finnfeeds Int Ltd | Use of an enzyme for manufacturing an agent for the treatment and/or prophylaxis of coccidiosis |
| GB9416841D0 (en) * | 1994-08-19 | 1994-10-12 | Finnfeeds Int Ltd | An enzyme feed additive and animal feed including it |
| CA2206992C (en) | 1994-12-09 | 2001-04-10 | The Procter & Gamble Company | Automatic dishwashing composition containing particles of diacyl peroxides |
| US5534302A (en) | 1995-01-05 | 1996-07-09 | National Science Council | Method of preparing a fiber reinforced modified phenolic resin composite |
| US5534179A (en) | 1995-02-03 | 1996-07-09 | Procter & Gamble | Detergent compositions comprising multiperacid-forming bleach activators |
| AR000862A1 (es) * | 1995-02-03 | 1997-08-06 | Novozymes As | Variantes de una ó-amilasa madre, un metodo para producir la misma, una estructura de adn y un vector de expresion, una celula transformada por dichaestructura de adn y vector, un aditivo para detergente, composicion detergente, una composicion para lavado de ropa y una composicion para la eliminacion del |
| US5780285A (en) * | 1995-03-03 | 1998-07-14 | Genentech, Inc. | Subtilisin variants capable of cleaving substrates containing dibasic residues |
| US5837516A (en) * | 1995-03-03 | 1998-11-17 | Genentech, Inc. | Subtilisin variants capable of cleaving substrates containing basic residues |
| US6455295B1 (en) * | 1995-03-08 | 2002-09-24 | The Procter & Gamble Company | Subtilisin Carlsberg variants having decreased adsorption and increased hydrolysis |
| IL117350A0 (en) * | 1995-03-09 | 1996-07-23 | Procter & Gamble | Proteinase k variants having decreased adsorption and increased hydrolysis |
| US6475765B1 (en) * | 1995-03-09 | 2002-11-05 | Procter & Gamble Company | Subtilisin DY variants having decreased adsorption and increased hydrolysis |
| IL117352A0 (en) * | 1995-03-09 | 1996-07-23 | Procter & Gamble | Thermitase variants having decreased adsorption and increased hydrolysis |
| US5523434A (en) | 1995-03-15 | 1996-06-04 | The Procter & Gamble Company | Synthesis of bleach activators |
| ATE195001T1 (de) * | 1995-04-17 | 2000-08-15 | Procter & Gamble | Herstellung und verwendung von diacylperoxid enthaltende kompositteilchen |
| US5837517A (en) * | 1995-05-05 | 1998-11-17 | Novo Nordisk A/S | Protease variants and compositions |
| AU5796296A (en) | 1995-06-16 | 1997-01-15 | Procter & Gamble Company, The | Bleach compositions comprising cobalt catalysts |
| US5597936A (en) | 1995-06-16 | 1997-01-28 | The Procter & Gamble Company | Method for manufacturing cobalt catalysts |
| DE69613842T2 (de) | 1995-06-16 | 2002-04-04 | The Procter & Gamble Company, Cincinnati | Maschinengeschirrspülmittel, die kobaltkatalysatoren enthalten |
| US5576282A (en) | 1995-09-11 | 1996-11-19 | The Procter & Gamble Company | Color-safe bleach boosters, compositions and laundry methods employing same |
| US5703034A (en) * | 1995-10-30 | 1997-12-30 | The Procter & Gamble Company | Bleach catalyst particles |
| US5762647A (en) * | 1995-11-21 | 1998-06-09 | The Procter & Gamble Company | Method of laundering with a low sudsing granular detergent composition containing optimally selected levels of a foam control agent bleach activator/peroxygen bleaching agent system and enzyme |
| WO1997022680A1 (en) * | 1995-12-20 | 1997-06-26 | The Procter & Gamble Company | Bleach catalyst plus enzyme particles |
| US5883065A (en) * | 1996-01-22 | 1999-03-16 | The Procter & Gamble Company | Phase separated detergent composition |
| DE69731282T2 (de) * | 1996-03-07 | 2006-03-02 | The Procter & Gamble Company, Cincinnati | Waschemittelzusammensetzungen mit proteasen und verbesserten amylasen |
| US6069122A (en) * | 1997-06-16 | 2000-05-30 | The Procter & Gamble Company | Dishwashing detergent compositions containing organic diamines for improved grease cleaning, sudsing, low temperature stability and dissolution |
| US6177392B1 (en) * | 1997-01-13 | 2001-01-23 | Ecolab Inc. | Stable solid block detergent composition |
| US6369011B1 (en) * | 1997-06-04 | 2002-04-09 | The Procter & Gamble Company | Protease enzymes for tough cleaning and/or spot and film reduction and compositions incorporating same |
| US6057171A (en) * | 1997-09-25 | 2000-05-02 | Frequency Technology, Inc. | Methods for determining on-chip interconnect process parameters |
| AR016969A1 (es) * | 1997-10-23 | 2001-08-01 | Procter & Gamble | VARIANTE DE PROTEASA, ADN, VECTOR DE EXPRESIoN, MICROORGANISMO HUESPED, COMPOSICIoN DE LIMPIEZA, ALIMENTO PARA ANIMALES Y COMPOSICIoN PARA TRATAR UN TEXTIL |
-
1998
- 1998-10-22 AR ARP980105280A patent/AR016969A1/es active IP Right Grant
- 1998-10-22 AR ARP980105279A patent/AR015977A1/es unknown
- 1998-10-22 MA MA25312A patent/MA24811A1/fr unknown
- 1998-10-22 MA MA25310A patent/MA25044A1/fr unknown
- 1998-10-23 CN CNB988126109A patent/CN1191347C/zh not_active Expired - Lifetime
- 1998-10-23 CA CA2307638A patent/CA2307638C/en not_active Expired - Fee Related
- 1998-10-23 BR BR9813879-0A patent/BR9813879A/pt not_active IP Right Cessation
- 1998-10-23 ID IDW20000966A patent/ID28256A/id unknown
- 1998-10-23 CN CNB988126370A patent/CN1198912C/zh not_active Expired - Lifetime
- 1998-10-23 AT AT05006461T patent/ATE513479T1/de active
- 1998-10-23 TR TR2000/01112T patent/TR200001112T2/xx unknown
- 1998-10-23 AU AU11996/99A patent/AU1199699A/en not_active Abandoned
- 1998-10-23 CZ CZ20001479A patent/CZ20001479A3/cs unknown
- 1998-10-23 AT AT98955109T patent/ATE399469T1/de active
- 1998-10-23 CN CN98811435A patent/CN1279721A/zh active Pending
- 1998-10-23 DE DE69839677T patent/DE69839677D1/de not_active Expired - Lifetime
- 1998-10-23 TR TR2001/01199T patent/TR200101199T2/xx unknown
- 1998-10-23 JP JP2000517091A patent/JP2001520046A/ja active Pending
- 1998-10-23 DE DE69839057T patent/DE69839057T2/de not_active Expired - Lifetime
- 1998-10-23 EP EP98956181A patent/EP1025241B2/en not_active Expired - Lifetime
- 1998-10-23 CN CN2010105436254A patent/CN102021159A/zh active Pending
- 1998-10-23 CA CA002306894A patent/CA2306894C/en not_active Expired - Fee Related
- 1998-10-23 PT PT98955109T patent/PT1027418E/pt unknown
- 1998-10-23 CZ CZ20001493A patent/CZ300347B6/cs not_active IP Right Cessation
- 1998-10-23 EP EP98955111A patent/EP1025240B1/en not_active Revoked
- 1998-10-23 ID IDW20000751D patent/ID25637A/id unknown
- 1998-10-23 KR KR1020007004391A patent/KR20010024555A/ko not_active Application Discontinuation
- 1998-10-23 CZ CZ20001503A patent/CZ301469B6/cs not_active IP Right Cessation
- 1998-10-23 ES ES98955079T patent/ES2319470T5/es not_active Expired - Lifetime
- 1998-10-23 PL PL340600A patent/PL191529B1/pl unknown
- 1998-10-23 CA CA2618389A patent/CA2618389C/en not_active Expired - Fee Related
- 1998-10-23 CA CA002306794A patent/CA2306794C/en not_active Expired - Fee Related
- 1998-10-23 WO PCT/US1998/022500 patent/WO1999020769A2/en active IP Right Grant
- 1998-10-23 PT PT05015947T patent/PT1624050E/pt unknown
- 1998-10-23 DK DK98955079.3T patent/DK1025194T4/da active
- 1998-10-23 CN CNB2004100399770A patent/CN1295313C/zh not_active Expired - Lifetime
- 1998-10-23 KR KR1020007004389A patent/KR20010024554A/ko not_active Application Discontinuation
- 1998-10-23 CA CA002672492A patent/CA2672492A1/en not_active Abandoned
- 1998-10-23 PT PT98956181T patent/PT1025241E/pt unknown
- 1998-10-23 TR TR2000/02013T patent/TR200002013T2/xx unknown
- 1998-10-23 AU AU11971/99A patent/AU742632B2/en not_active Ceased
- 1998-10-23 BR BR9813260-1A patent/BR9813260A/pt not_active IP Right Cessation
- 1998-10-23 NZ NZ520771A patent/NZ520771A/en not_active IP Right Cessation
- 1998-10-23 EP EP05018139A patent/EP1612271B1/en not_active Revoked
- 1998-10-23 DK DK98956181.6T patent/DK1025241T4/da active
- 1998-10-23 TR TR2000/01113T patent/TR200001113T2/xx unknown
- 1998-10-23 US US09/178,173 patent/US6482628B1/en not_active Expired - Lifetime
- 1998-10-23 ID IDW20000749A patent/ID25897A/id unknown
- 1998-10-23 CA CA002308206A patent/CA2308206C/en not_active Expired - Fee Related
- 1998-10-23 CA CA2308113A patent/CA2308113C/en not_active Expired - Fee Related
- 1998-10-23 US US09/178,155 patent/US6312936B1/en not_active Expired - Lifetime
- 1998-10-23 CA CA002672500A patent/CA2672500A1/en not_active Abandoned
- 1998-10-23 JP JP2000517049A patent/JP2001520307A/ja active Pending
- 1998-10-23 ID IDW20000967A patent/ID26501A/id unknown
- 1998-10-23 TR TR2001/01861T patent/TR200101861T2/xx unknown
- 1998-10-23 JP JP2000517050A patent/JP2001520308A/ja active Pending
- 1998-10-23 ES ES98955109T patent/ES2310014T5/es not_active Expired - Lifetime
- 1998-10-23 WO PCT/US1998/022590 patent/WO1999020771A2/en active IP Right Grant
- 1998-10-23 AU AU11969/99A patent/AU742573B2/en not_active Ceased
- 1998-10-23 AT AT98955079T patent/ATE417099T1/de active
- 1998-10-23 DE DE69840335T patent/DE69840335D1/de not_active Expired - Lifetime
- 1998-10-23 EP EP05006461A patent/EP1571199B1/en not_active Expired - Lifetime
- 1998-10-23 DK DK98953936T patent/DK1025239T3/da active
- 1998-10-23 DK DK05018139.5T patent/DK1612271T3/da active
- 1998-10-23 EP EP98955109A patent/EP1027418B2/en not_active Expired - Lifetime
- 1998-10-23 PT PT05006461T patent/PT1571199E/pt unknown
- 1998-10-23 NZ NZ520770A patent/NZ520770A/en not_active IP Right Cessation
- 1998-10-23 BR BRPI9813266A patent/BRPI9813266B1/pt not_active IP Right Cessation
- 1998-10-23 AU AU11994/99A patent/AU742694B2/en not_active Ceased
- 1998-10-23 WO PCT/US1998/022486 patent/WO1999020723A2/en not_active Application Discontinuation
- 1998-10-23 CZ CZ20001478A patent/CZ20001478A3/cs unknown
- 1998-10-23 KR KR1020007004385A patent/KR100616143B1/ko not_active IP Right Cessation
- 1998-10-23 PT PT05018139T patent/PT1612271E/pt unknown
- 1998-10-23 WO PCT/US1998/022572 patent/WO1999020770A2/en not_active Application Discontinuation
- 1998-10-23 DK DK98955109.8T patent/DK1027418T4/da active
- 1998-10-23 AT AT98956181T patent/ATE292179T1/de active
- 1998-10-23 PT PT03024798T patent/PT1398367E/pt unknown
- 1998-10-23 ES ES98953936T patent/ES2301214T3/es not_active Expired - Lifetime
- 1998-10-23 JP JP2000517090A patent/JP4511025B2/ja not_active Expired - Fee Related
- 1998-10-23 DK DK05015947.4T patent/DK1624050T3/da active
- 1998-10-23 AU AU11181/99A patent/AU758371B2/en not_active Ceased
- 1998-10-23 PT PT98953936T patent/PT1025239E/pt unknown
- 1998-10-23 NZ NZ519204A patent/NZ519204A/en not_active IP Right Cessation
- 1998-10-23 CN CNB988114364A patent/CN100342016C/zh not_active Expired - Fee Related
- 1998-10-23 BR BRPI9813115-0A patent/BRPI9813115B1/pt not_active IP Right Cessation
- 1998-10-23 KR KR1020007004382A patent/KR100612981B1/ko not_active IP Right Cessation
- 1998-10-23 TR TR2000/02014T patent/TR200002014T2/xx unknown
- 1998-10-23 US US09/177,353 patent/US6673590B1/en not_active Expired - Lifetime
- 1998-10-23 AT AT05015947T patent/ATE445002T1/de active
- 1998-10-23 HU HU0103080A patent/HUP0103080A2/hu unknown
- 1998-10-23 AT AT05018139T patent/ATE511540T1/de active
- 1998-10-23 AR ARP980105303A patent/AR013718A1/es unknown
- 1998-10-23 TR TR2000/01111T patent/TR200001111T2/xx unknown
- 1998-10-23 CN CNB988113139A patent/CN1195857C/zh not_active Expired - Fee Related
- 1998-10-23 JP JP2000517046A patent/JP2001520305A/ja active Pending
- 1998-10-23 EP EP98955081A patent/EP1082404A2/en not_active Ceased
- 1998-10-23 DK DK05006461.7T patent/DK1571199T3/da active
- 1998-10-23 EP EP98955079A patent/EP1025194B2/en not_active Expired - Lifetime
- 1998-10-23 CN CNB988126400A patent/CN100558871C/zh not_active Expired - Fee Related
- 1998-10-23 DE DE69841226T patent/DE69841226D1/de not_active Expired - Lifetime
- 1998-10-23 DK DK98955111T patent/DK1025240T3/da active
- 1998-10-23 CN CNB2004100399766A patent/CN100506963C/zh not_active Expired - Fee Related
- 1998-10-23 CZ CZ20001477A patent/CZ20001477A3/cs unknown
- 1998-10-23 AR ARP980105305A patent/AR013719A1/es not_active Application Discontinuation
- 1998-10-23 AT AT98955111T patent/ATE314478T1/de active
- 1998-10-23 TR TR2000/02057T patent/TR200002057T2/xx unknown
- 1998-10-23 HU HU0100859A patent/HUP0100859A2/hu unknown
- 1998-10-23 EP EP05015947A patent/EP1624050B9/en not_active Expired - Lifetime
- 1998-10-23 AT AT98953936T patent/ATE384799T1/de active
- 1998-10-23 DE DE69829577T patent/DE69829577T3/de not_active Expired - Lifetime
- 1998-10-23 PT PT98955079T patent/PT1025194E/pt unknown
- 1998-10-23 ES ES98955111T patent/ES2255189T3/es not_active Expired - Lifetime
- 1998-10-23 AT AT03024798T patent/ATE416624T1/de active
- 1998-10-23 DE DE69840346T patent/DE69840346D1/de not_active Expired - Lifetime
- 1998-10-23 KR KR1020007004380A patent/KR100564039B1/ko not_active IP Right Cessation
- 1998-10-23 EP EP03024798A patent/EP1398367B1/en not_active Expired - Lifetime
- 1998-10-23 KR KR1020007004388A patent/KR20010031380A/ko active IP Right Grant
- 1998-10-23 HU HU0104539A patent/HUP0104539A3/hu unknown
- 1998-10-23 BR BR9815230-0A patent/BR9815230A/pt not_active Application Discontinuation
- 1998-10-23 JP JP2000517089A patent/JP4346237B2/ja not_active Expired - Fee Related
- 1998-10-23 CA CA2307640A patent/CA2307640C/en not_active Expired - Fee Related
- 1998-10-23 CZ CZ20001504A patent/CZ302287B6/cs not_active IP Right Cessation
- 1998-10-23 DE DE69833015T patent/DE69833015T2/de not_active Expired - Lifetime
- 1998-10-23 ES ES05015947T patent/ES2332915T3/es not_active Expired - Lifetime
- 1998-10-23 DK DK03024798T patent/DK1398367T3/da active
- 1998-10-23 ES ES03024798T patent/ES2319401T3/es not_active Expired - Lifetime
- 1998-10-23 AU AU12762/99A patent/AU1276299A/en not_active Abandoned
- 1998-10-23 ES ES98956181T patent/ES2241180T5/es not_active Expired - Lifetime
- 1998-10-23 BR BR9814097-3A patent/BR9814097A/pt not_active Application Discontinuation
- 1998-10-23 WO PCT/US1998/022482 patent/WO1999020726A1/en not_active Application Discontinuation
- 1998-10-23 EP EP98953936A patent/EP1025239B1/en not_active Expired - Lifetime
- 1998-10-24 EG EG129398A patent/EG21711A/xx active
- 1998-10-24 EG EG129498A patent/EG22075A/xx active
-
1999
- 1999-01-14 TW TW087117614A patent/TW585912B/zh not_active IP Right Cessation
- 1999-01-18 TW TW087117606A patent/TW562859B/zh not_active IP Right Cessation
-
2000
- 2000-04-12 NO NO20001899A patent/NO327607B1/no not_active IP Right Cessation
- 2000-04-12 NO NO20001901A patent/NO20001901L/no not_active Application Discontinuation
- 2000-04-12 NO NO20001900A patent/NO326992B1/no not_active IP Right Cessation
-
2001
- 2001-11-02 AU AU87337/01A patent/AU8733701A/en not_active Abandoned
- 2001-11-02 US US10/033,325 patent/US6815193B2/en not_active Expired - Lifetime
-
2002
- 2002-02-01 HK HK02100781.1A patent/HK1039159B/zh not_active IP Right Cessation
- 2002-08-27 US US10/228,572 patent/US6927055B2/en not_active Expired - Fee Related
-
2006
- 2006-01-11 AR ARP060100112A patent/AR052355A2/es not_active Application Discontinuation
- 2006-01-11 AR ARP060100108A patent/AR052353A2/es active IP Right Grant
- 2006-01-11 AR ARP060100109A patent/AR052282A2/es not_active Application Discontinuation
- 2006-01-11 AR ARP060100107A patent/AR052281A2/es not_active Application Discontinuation
- 2006-01-11 AR ARP060100111A patent/AR052354A2/es not_active Application Discontinuation
- 2006-01-11 AR ARP060100110A patent/AR052453A2/es unknown
-
2012
- 2012-06-20 JP JP2012138496A patent/JP5612026B2/ja not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109312323A (zh) * | 2016-06-09 | 2019-02-05 | 花王株式会社 | 突变碱性蛋白酶 |
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100342016C (zh) | 多点置换的蛋白酶变体 | |
| CN1056649C (zh) | 枯草杆菌蛋白酶变异体 | |
| EP2385112B1 (en) | Subtilase variants | |
| JP6792524B2 (ja) | 複数置換プロテアーゼ変異体 | |
| RU2252254C2 (ru) | Вариант субтилизина bacillus (варианты), кодирующая его днк, экспрессирующий вектор и очищающая композиция | |
| CN1711354A (zh) | 枯草杆菌酶变体 |
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 | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1039159 Country of ref document: HK |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20071010 Termination date: 20161023 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |