RU2016102045A - Варианты субтилаз и кодирующие их полинуклеотиды - Google Patents
Варианты субтилаз и кодирующие их полинуклеотиды Download PDFInfo
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- RU2016102045A RU2016102045A RU2016102045A RU2016102045A RU2016102045A RU 2016102045 A RU2016102045 A RU 2016102045A RU 2016102045 A RU2016102045 A RU 2016102045A RU 2016102045 A RU2016102045 A RU 2016102045A RU 2016102045 A RU2016102045 A RU 2016102045A
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- 101710135785 Subtilisin-like protease Proteins 0.000 title claims 8
- 108091033319 polynucleotide Proteins 0.000 title claims 5
- 102000040430 polynucleotide Human genes 0.000 title claims 5
- 239000002157 polynucleotide Substances 0.000 title claims 5
- 229920001184 polypeptide Polymers 0.000 claims 15
- 102000004196 processed proteins & peptides Human genes 0.000 claims 15
- 108090000765 processed proteins & peptides Proteins 0.000 claims 15
- 108091005804 Peptidases Proteins 0.000 claims 6
- 239000004365 Protease Substances 0.000 claims 6
- 239000003599 detergent Substances 0.000 claims 6
- 239000000203 mixture Substances 0.000 claims 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 5
- 108091026890 Coding region Proteins 0.000 claims 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims 3
- 238000004140 cleaning Methods 0.000 claims 2
- 230000000295 complement effect Effects 0.000 claims 2
- 238000000034 method Methods 0.000 claims 2
- 238000006467 substitution reaction Methods 0.000 claims 2
- 238000005406 washing Methods 0.000 claims 2
- 108010065511 Amylases Proteins 0.000 claims 1
- 102000013142 Amylases Human genes 0.000 claims 1
- 102100032487 Beta-mannosidase Human genes 0.000 claims 1
- 102000016938 Catalase Human genes 0.000 claims 1
- 108010053835 Catalase Proteins 0.000 claims 1
- 108010059892 Cellulase Proteins 0.000 claims 1
- 108010084185 Cellulases Proteins 0.000 claims 1
- 102000005575 Cellulases Human genes 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- 108090001060 Lipase Proteins 0.000 claims 1
- 102000004882 Lipase Human genes 0.000 claims 1
- 239000004367 Lipase Substances 0.000 claims 1
- 108010029182 Pectin lyase Proteins 0.000 claims 1
- 102000035195 Peptidases Human genes 0.000 claims 1
- 108700020962 Peroxidase Proteins 0.000 claims 1
- 102000003992 Peroxidases Human genes 0.000 claims 1
- 108010059820 Polygalacturonase Proteins 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 claims 1
- 235000019418 amylase Nutrition 0.000 claims 1
- 229940025131 amylases Drugs 0.000 claims 1
- 108010055059 beta-Mannosidase Proteins 0.000 claims 1
- 108010005400 cutinase Proteins 0.000 claims 1
- 238000012217 deletion Methods 0.000 claims 1
- 230000037430 deletion Effects 0.000 claims 1
- 238000004851 dishwashing Methods 0.000 claims 1
- 238000010573 double replacement reaction Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 claims 1
- 108010093305 exopolygalacturonase Proteins 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 239000000499 gel Substances 0.000 claims 1
- 239000008187 granular material Substances 0.000 claims 1
- 229910052736 halogen Inorganic materials 0.000 claims 1
- 150000002367 halogens Chemical class 0.000 claims 1
- 238000003780 insertion Methods 0.000 claims 1
- 230000037431 insertion Effects 0.000 claims 1
- 235000019421 lipase Nutrition 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 239000006072 paste Substances 0.000 claims 1
- 108010023506 peroxygenase Proteins 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- -1 xanthanases Proteins 0.000 claims 1
- 108010083879 xyloglucan endo(1-4)-beta-D-glucanase Proteins 0.000 claims 1
Classifications
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- 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
-
- 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
- C11D3/38681—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)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
-
- 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)
-
- 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)
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
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- Enzymes And Modification Thereof (AREA)
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Claims (116)
1. Вариант субтилазы, обладающий протеазной активностью и содержащий в себе двойную замену 194P+261D, и где положения соответствуют положениям в зрелом полипептиде с SEQ ID NO: 2.
2. Вариант по п. 1, который дополнительно содержит одно или несколько изменений из группы, состоящей из 3{F, I, L, V, Y}, 9{A, G, M, T}, 18{A, D, E, G, M, S, T}, 40{D, E}, 43{D, E}, 45{D, E}, 58{F, Y}, 59{D, E}, 61{D, E}, 72{L, V}, 76{D, E}, 99{A, D, E, M, T}, 104{F, Y}, 105{D, E}, 120{D, E, I, L, V}, 130*, 133{D, E}, 141{F, Y}, 182{D, E}, 183{D, E}, 184{D, E}, 205{I, L}, 206{D, E}, 212{D, E}, 218{D, E}, 224{A, G, M, S}, 225{A, G, M, S, T}, 228{G, M, S, T}, 236{D, E}, 245{H, K}, 259{D, E} и 262{F, W, Y}.
3. Вариант по п. 1 или 2, где двойная замена представляет собой A194P+N261D.
4. Вариант по любому из пп. 1-3, где вариантная субтилаза представляет собой:
а) полипептид, последовательность которого по меньшей мере на 60%, например, по меньшей мере на 65%, по меньшей мере на 70%, по меньшей мере на 75%, по меньшей мере на 80%, по меньшей мере на 85%, по меньшей мере на 90%, по меньшей мере на 95%, по меньшей мере на 96%, по меньшей мере на 97%, по меньшей мере на 98%, но меньше чем на 100%, идентична последовательности зрелого полипептида исходной субтилазы;
b) полипептид, кодируемый полинуклеотидом, который гибридизуется в условиях низкой жесткости, в условиях средней жесткости, условиях умеренно высокой жесткости, условиях высокой жесткости или условиях очень высокой жесткости с
(i) кодирующей последовательностью зрелого полипептида исходной субтилазы или
(ii) полноразмерной последовательностью, комплементарной (i); или
с) полипептид, кодируемый полинуклеотидом, последовательность которого по меньшей мере на 60%, например, по меньшей мере на 65%, по меньшей мере на 70%, по меньшей мере на 75%, по меньшей мере на 80%, по меньшей мере на 85%, по меньшей мере на 90%, по меньшей мере на 95%, по меньшей мере на 96%, по меньшей мере на 97%, по меньшей мере на 98%, но меньше чем на 100%, идентична кодирующей последовательности зрелого полипептида исходной субтилазы.
5. Вариант по любому из пп. 1-4, где вариантная субтилаза представляет собой:
а) полипептид, последовательность которого по меньшей мере на 60%, например, по меньшей мере на 65%, по меньшей мере на 70%, по меньшей мере на 75%, по меньшей мере на 80%, по меньшей мере на 85%, по меньшей мере на 90%, по меньшей мере на 95%, по меньшей мере на 96%, по меньшей мере на 97%, по меньшей мере на 98%, но меньше чем на 100%, идентична последовательности зрелого полипептида с SEQ ID NO: 4;
b) полипептид, кодируемый полинуклеотидом, который гибридизуется в условиях низкой жесткости, в условиях средней жесткости, условиях умеренно высокой жесткости, условиях высокой жесткости или условиях очень высокой жесткости с
(i) кодирующей последовательностью зрелого полипептида с SEQ ID NO: 3 или
(ii) полноразмерной последовательностью, комплементарной (i); или
c) полипептид, кодируемый полинуклеотидом, последовательность которого по меньшей мере на 60%, например, по меньшей мере на 65%, по меньшей мере на 70%, по меньшей мере на 75%, по меньшей мере на 80%, по меньшей мере на 85%, по меньшей мере на 90%, по меньшей мере на 95%, по меньшей мере на 96%, по меньшей мере на 97%, по меньшей мере на 98%, но меньше чем на 100%, идентична кодирующей последовательности зрелого полипептида с SEQ ID NO: 3.
6. Вариант по любому из предшествующих пунктов, где общее число изменений составляет от 3 до 30, предпочтительно от 3 до 20, более предпочтительно от 3 до 15, и даже более предпочтительно от 3 до 10, особенно предпочтительно от 3 до 8 изменений.
7. Вариант по любому из предшествующих пунктов, где общее число изменений составляет 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 или 30 изменений.
8. Вариант по любому из предшествующих пунктов, где вариант состоит из 150-350, например, 175-330, 200-310, 220-300, 240-290, 260-280 или 269-275 аминокислот.
9. Вариант по любому из предшествующих пунктов, где вариант содержит одно или несколько изменений, выбранных из группы, состоящей из:
3F+194P+261D;
3I+194P+261D;
3L+194P+261D;
3V+194P+261D;
3Y+194P+261D;
9A+194P+261D;
9G+194P+261D;
9M+194P+261D;
9T+194P+261D;
9+194P+261D;
18A+194P+261D;
18D+194P+261D;
18E+194P+261D;
18G+194P+261D;
18M+194P+261D;
18S+194P+261D;
18T+194P+261D;
40D+194P+261D;
40E+194P+261D;
43D+194P+261D;
43E+194P+261D;
45D+194P+261D;
45E+194P+261D;
58F+194P+261D;
58Y+194P+261D;
59D+194P+261D;
59E+194P+261D;
61D+194P+261D;
61E+194P+261D;
72L+194P+261D;
72V+194P+261D;
76D+194P+261D;
76E+194P+261D;
99A+194P+261D;
99D+194P+261D;
99E+194P+261D;
99M+194P+261D;
99T+194P+261D;
104F+194P+261D;
104Y+194P+261D;
105D+194P+261D;
105E+194P+261D;
120D+194P+261D;
120E+194P+261D;
120I+194P+261D;
120L+194P+261D;
120V+194P+261D;
130*+194P+261D;
133D+194P+261D;
133E+194P+261D;
141F+194P+261D;
141Y+194P+261D;
182D+194P+261D;
182E+194P+261D;
183D+194P+261D;
183E+194P+261D;
184D+194P+261D;
184E+194P+261D;
194P+205I+261D;
194P+205L+261D;
194P+206D+261D;
194P+206E+261D;
194P+212D+261D;
194P+212E+261D;
194P+218D+261D;
194P+218E+261D;
194P+224A+261D;
194P+224G+261D;
194P+224M+261D;
194P+224S+261D;
194P+225A+261D;
194P+225G+261D;
194P+225M+261D;
194P+225S+261D;
194P+225T+261D;
194P+228G+261D;
194P+228M+261D;
194P+228S+261D;
194P+228T+261D;
194P+236D+261D;
194P+236E+261D;
194P+245H+261D;
194P+245K+261D;
194P+259D+261D;
194P+259E+261D;
194P+261D+262F;
194P+261D+262W; и
194P+261D+262Y.
10. Вариант по любому из пп. 1-9, содержащий замену, делецию и/или вставку в одном или нескольких положениях (например, нескольких), выбранных из группы, состоящей из положений: 4, 9, 12, 14, 15, 40, 63, 68, 79, 84, 86, 88, 92, 98, 101, 132, 146, 188, 217, 245, 255 и 270, где каждое положение соответствует положению в зрелом полипептиде с SEQ ID NO: 2.
11. Вариант по любому из пп. 1-10, который обладает улучшенной стабильностью, предпочтительно - улучшенной стабильностью при мытье, по сравнению с исходной протеазой или по сравнению с эталонной протеазой.
12. Вариант по п. 11, который обладает улучшенной моющей способностью по сравнению с исходной протеазой или по сравнению с эталонной протеазой.
13. Композиция моющего средства, содержащая вариант согласно любому из пп. 1-12.
14. Композиция моющего средства по п. 13, дополнительно содержащая один или несколько компонентов моющего средства.
15. Композиция моющего средства согласно любому из пп. 13-14, дополнительно содержащая один или несколько дополнительных ферментов, выбранных из группы, состоящей из протеаз, амилаз, липаз, кутиназ, целлюлаз, эндоглюканаз, ксилоглюканаз, пектиназ, пектин-лиаз, ксантаназ, пероксидаз, галогенпероксигеназ, каталаз и маннаназ или любой их смеси.
16. Композиция моющего средства согласно любому из пп. 13-15 в форме бруска, гомогенной таблетки, таблетки с двумя или более слоями, пакетика с одной или несколькими камерами, обычного или уплотненного порошка, гранулы, пасты, геля или обычной, плотной или концентрированной жидкости.
17. Применение композиции моющего средства согласно любому из пп. 13-16 в способе очистки, таком как стирка или очистка твердой поверхности, такая как мытье посуды.
18. Способ получения варианта субтилазы, включающий в себя (a) ведение в исходную субтилазу двойной замены 194P+261D и одного или нескольких изменений из группы, состоящей из 3{F, I, L, V, Y}, 9{A, G, M, T}, 18{A, D, E, G, M, S, T}, 40{D, E}, 43{D, E}, 45{D, E}, 58{F, Y}, 59{D, E}, 61{D, E}, 72{L, V}, 76{D, E}, 99{A, D, E, M, T}, 104{F, Y}, 105{D, E}, 120{D, E, I, L, V}, 130*, 133{D, E}, 141{F, Y}, 182{D, E}, 183{D, E}, 184{D, E}, 205{I, L}, 206{D, E}, 212{D, E}, 218{D, E}, 224{A, G, M, S}, 225{A, G, M, S, T}, 228{G, M, S, T}, 236{D, E}, 245{H, K}, 259{D, E} и 262{F, W, Y}, где каждое положение соответствует положению в зрелом полипептиде с SEQ ID NO: 2 и (b) извлечение варианта.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP13174065.6 | 2013-06-27 | ||
| EP13174065 | 2013-06-27 | ||
| PCT/EP2014/063749 WO2014207227A1 (en) | 2013-06-27 | 2014-06-27 | Subtilase variants and polynucleotides encoding same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| RU2016102045A true RU2016102045A (ru) | 2017-08-01 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| RU2016102045A RU2016102045A (ru) | 2013-06-27 | 2014-06-27 | Варианты субтилаз и кодирующие их полинуклеотиды |
Country Status (7)
| Country | Link |
|---|---|
| US (2) | US10378001B2 (ru) |
| EP (1) | EP3013956B1 (ru) |
| CN (1) | CN105874067A (ru) |
| BR (1) | BR112015032524A2 (ru) |
| FI (1) | FI3013956T3 (ru) |
| RU (1) | RU2016102045A (ru) |
| WO (1) | WO2014207227A1 (ru) |
Families Citing this family (43)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017089093A1 (en) * | 2015-11-25 | 2017-06-01 | Unilever N.V. | A liquid detergent composition |
| DE102017125560A1 (de) | 2017-11-01 | 2019-05-02 | Henkel Ag & Co. Kgaa | Reinigungszusammensetzungen, die dispersine iii enthalten |
| US12012573B2 (en) | 2018-07-02 | 2024-06-18 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020008024A1 (en) | 2018-07-06 | 2020-01-09 | Novozymes A/S | Cleaning compositions and uses thereof |
| WO2020070249A1 (en) | 2018-10-03 | 2020-04-09 | Novozymes A/S | Cleaning compositions |
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-
2014
- 2014-06-27 US US14/899,583 patent/US10378001B2/en active Active
- 2014-06-27 CN CN201480040367.4A patent/CN105874067A/zh active Pending
- 2014-06-27 BR BR112015032524A patent/BR112015032524A2/pt not_active Application Discontinuation
- 2014-06-27 EP EP14733216.7A patent/EP3013956B1/en active Active
- 2014-06-27 RU RU2016102045A patent/RU2016102045A/ru not_active Application Discontinuation
- 2014-06-27 FI FIEP14733216.7T patent/FI3013956T3/fi active
- 2014-06-27 WO PCT/EP2014/063749 patent/WO2014207227A1/en not_active Ceased
-
2019
- 2019-06-27 US US16/454,226 patent/US20190338266A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| CN105874067A (zh) | 2016-08-17 |
| EP3013956B1 (en) | 2023-03-01 |
| US20190338266A1 (en) | 2019-11-07 |
| BR112015032524A2 (pt) | 2017-08-29 |
| US10378001B2 (en) | 2019-08-13 |
| US20160137998A1 (en) | 2016-05-19 |
| WO2014207227A1 (en) | 2014-12-31 |
| FI3013956T3 (fi) | 2023-05-23 |
| EP3013956A1 (en) | 2016-05-04 |
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