RU2012133453A - Альфа-амилазы - Google Patents
Альфа-амилазы Download PDFInfo
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- RU2012133453A RU2012133453A RU2012133453/10A RU2012133453A RU2012133453A RU 2012133453 A RU2012133453 A RU 2012133453A RU 2012133453/10 A RU2012133453/10 A RU 2012133453/10A RU 2012133453 A RU2012133453 A RU 2012133453A RU 2012133453 A RU2012133453 A RU 2012133453A
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- 108090000637 alpha-Amylases Proteins 0.000 claims abstract 29
- 102000004139 alpha-Amylases Human genes 0.000 claims abstract 28
- 229940024171 alpha-amylase Drugs 0.000 claims abstract 28
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract 3
- 229910052791 calcium Inorganic materials 0.000 claims abstract 3
- 239000011575 calcium Substances 0.000 claims abstract 3
- 238000000034 method Methods 0.000 claims 5
- 229920002472 Starch Polymers 0.000 claims 4
- 238000000855 fermentation Methods 0.000 claims 4
- 230000004151 fermentation Effects 0.000 claims 4
- 235000019698 starch Nutrition 0.000 claims 4
- 239000008107 starch Substances 0.000 claims 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 3
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 claims 2
- 102100022624 Glucoamylase Human genes 0.000 claims 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims 2
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims 2
- 150000007523 nucleic acids Chemical group 0.000 claims 2
- 239000000126 substance Substances 0.000 claims 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 claims 2
- 235000000346 sugar Nutrition 0.000 claims 2
- 150000008163 sugars Chemical class 0.000 claims 2
- VBUYCZFBVCCYFD-JJYYJPOSSA-M 2-dehydro-D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)C([O-])=O VBUYCZFBVCCYFD-JJYYJPOSSA-M 0.000 claims 1
- VBUYCZFBVCCYFD-NUNKFHFFSA-N 2-dehydro-L-idonic acid Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)C(=O)C(O)=O VBUYCZFBVCCYFD-NUNKFHFFSA-N 0.000 claims 1
- 108010011619 6-Phytase Proteins 0.000 claims 1
- 108010059892 Cellulase Proteins 0.000 claims 1
- 108010008885 Cellulose 1,4-beta-Cellobiosidase Proteins 0.000 claims 1
- VBUYCZFBVCCYFD-UHFFFAOYSA-N D-arabino-2-Hexulosonic acid Natural products OCC(O)C(O)C(O)C(=O)C(O)=O VBUYCZFBVCCYFD-UHFFFAOYSA-N 0.000 claims 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 claims 1
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- 108010028688 Isoamylase Proteins 0.000 claims 1
- 102000004195 Isomerases Human genes 0.000 claims 1
- 108090000769 Isomerases Proteins 0.000 claims 1
- 102000004882 Lipase Human genes 0.000 claims 1
- 108090001060 Lipase Proteins 0.000 claims 1
- 239000004367 Lipase Substances 0.000 claims 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims 1
- 239000004472 Lysine Substances 0.000 claims 1
- 108091005804 Peptidases Proteins 0.000 claims 1
- 102000035195 Peptidases Human genes 0.000 claims 1
- 239000004365 Protease Substances 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 150000007513 acids Chemical class 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 claims 1
- 108010019077 beta-Amylase Proteins 0.000 claims 1
- 108010047754 beta-Glucosidase Proteins 0.000 claims 1
- 102000006995 beta-Glucosidase Human genes 0.000 claims 1
- 229940106157 cellulase Drugs 0.000 claims 1
- 239000003599 detergent Substances 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 claims 1
- 239000012634 fragment Substances 0.000 claims 1
- 229940050410 gluconate Drugs 0.000 claims 1
- 235000011187 glycerol Nutrition 0.000 claims 1
- 229940059442 hemicellulase Drugs 0.000 claims 1
- 108010002430 hemicellulase Proteins 0.000 claims 1
- 239000000543 intermediate Substances 0.000 claims 1
- 239000004310 lactic acid Substances 0.000 claims 1
- 235000014655 lactic acid Nutrition 0.000 claims 1
- 235000019421 lipase Nutrition 0.000 claims 1
- -1 lysine) Chemical class 0.000 claims 1
- 150000007524 organic acids Chemical class 0.000 claims 1
- 235000005985 organic acids Nutrition 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 claims 1
- 239000000758 substrate Substances 0.000 claims 1
- 239000001384 succinic acid Substances 0.000 claims 1
- 150000005846 sugar alcohols Chemical class 0.000 claims 1
- 239000004094 surface-active agent Substances 0.000 claims 1
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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)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
- C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
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- 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
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- 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
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- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
- C12N15/8257—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
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- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
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- C12P7/14—Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
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- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01001—Alpha-amylase (3.2.1.1)
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Abstract
1. Выделенная альфа-амилаза, содержащая A- и C-домены чувствительной к кальцию альфа-амилазы и B-домен нечувствительной к кальцию альфа-амилазы.2. Выделенная альфа-амилаза, содержащая A-домен с по меньшей мере 60% идентичностью последовательностей с A-доменом с любой из SEQ ID NO. 1-12, 29 и 30, B-домен с по меньшей мере 60% идентичностью последовательностей с B-доменом с любой из SEQ ID NO. 13-16 и 31 и C-домен с по меньшей мере 60% идентичностью последовательностей с C-доменом с любой из SEQ ID NO. 1-12, 29 и 30.3. Альфа-амилаза по п.2, где A-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с A-доменом SEQ ID NO. 4.4. Альфа-амилаза по любому из пп.1-3, где A1-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с последовательностью, которая начинается в положении в диапазоне положений 1-5 и заканчивается в положении в диапазоне положений 94-114, например, начинается в положении в диапазоне положений 1-3 и заканчивается в положении в диапазоне положений 99-104 или начинается в положении в д
Claims (18)
1. Выделенная альфа-амилаза, содержащая A- и C-домены чувствительной к кальцию альфа-амилазы и B-домен нечувствительной к кальцию альфа-амилазы.
2. Выделенная альфа-амилаза, содержащая A-домен с по меньшей мере 60% идентичностью последовательностей с A-доменом с любой из SEQ ID NO. 1-12, 29 и 30, B-домен с по меньшей мере 60% идентичностью последовательностей с B-доменом с любой из SEQ ID NO. 13-16 и 31 и C-домен с по меньшей мере 60% идентичностью последовательностей с C-доменом с любой из SEQ ID NO. 1-12, 29 и 30.
3. Альфа-амилаза по п.2, где A-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с A-доменом SEQ ID NO. 4.
4. Альфа-амилаза по любому из пп.1-3, где A1-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с последовательностью, которая начинается в положении в диапазоне положений 1-5 и заканчивается в положении в диапазоне положений 94-114, например, начинается в положении в диапазоне положений 1-3 и заканчивается в положении в диапазоне положений 99-104 или начинается в положении в диапазоне положений 1-3 и заканчивается в положении в диапазоне положений 104-109, в частности, положения 1-104 SEQ ID NO. 4.
5. Альфа-амилаза по любому из пп.1-3, где B-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с последовательностью, которая начинается в положении в диапазоне положений 93-113 и заканчивается в положении в диапазоне положений 195-215, например, начинается в положении в диапазоне положений 97-109 и заканчивается в положении в диапазоне положений 199-211 или начинается в положении в диапазоне положений 100-106 и заканчивается в положении в диапазоне положений 202-208, в частности, положения 103-208 SEQ ID NO. 13.
6. Альфа-амилаза по любому из пп.1-3, где C-домен обладает по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с C-доменом SEQ ID NO. 4.
7. Альфа-амилаза по любому из пп.1-3, где A2- и C-домены обладают по меньшей мере 60% идентичностью последовательностей, например, по меньшей мере 65%, по меньшей мере 70%, по меньшей мере 75%, по меньшей мере 80%, по меньшей мере 85%, по меньшей мере 90%, по меньшей мере 91%, по меньшей мере 92%, по меньшей мере 93%, по меньшей мере 94%, по меньшей мере 95%, по меньшей мере 96%, по меньшей мере 97%, по меньшей мере 98%, по меньшей мере 99% или 100% идентичностью последовательностей с последовательностью, которая начинается в положении в диапазоне положений 201-221 и заканчивается в положении в диапазоне положений 478-483, например, начинается в положении в диапазоне положений 206-211 и заканчивается в положении в диапазоне положений 480-483 или начинается в положении в диапазоне положений 211-216 и заканчивается в положении в диапазоне положений 480-483, в частности, положения 211-483 SEQ ID NO. 4.
8. Альфа-амилаза по любому из пп.1-3, которая более термостабильна, чем альфа-амилаза с любой из SEQ ID NO. 1-12, 29 и 30.
9. Альфа-амилаза по любому из пп.1-3, которая обладает пониженной чувствительностью к кальцию по сравнению с альфа-амилазой с любой из SEQ ID NO. 1-12, 29 и 30.
10. Моющая композиция, содержащая альфа-амилазу по любому из пп.1-9 и поверхностно-активное вещество.
11. Композиция, содержащая альфа-амилазу по любому из пп.1-9 и один или более ферментов, выбранных из группы, состоящей из бета-амилазы, целлюлазы (бета-глюкозидазы, целлобиогидролазы и эндоглюказаназы) глюкоамилазы, гемицеллюлазы (например, ксиланазы), изоамилазы, изомеразы, липазы, фитазы, протеазы и пуллуланазы.
12. Способ получения разжиженного крахмала, включающий разжижение содержащего крахмал вещества с использованием альфа-амилазы по любому из пп.1-9.
13. Способ получения продукта ферментации, включающий
(a) разжижение содержащего крахмал вещества с использованием альфа-амилазы по любому из пп.1-9 для получения разжиженного затора;
(b) осахаривание разжиженного затора для получения сбраживаемых сахаров; и
(c) ферментацию сбраживаемых сахаров в присутствие ферментирующего организма.
14. Способ получения продукта ферментации, включающий приведение крахмального субстрата в контакт с альфа-амилазой по любому из пп.1-9, глюкоамилазой и ферментирующим организмом.
15. Способ по п.13 или 14, где продукт ферментации выбирают из группы, состоящей из спирта (например, этанол и бутанол), органических кислот (например, янтарная кислота и молочная кислота), сахароспиртов (например, глицерин), промежуточных соединений аскорбиновой кислоты (например, глюконат, 2-кето-D-глюконат, 2,5-дикето-D-глюконат и 2-кето-L-гулоновая кислота), аминокислот (например, лизин), белков (например, антитела и их фрагменты).
16. Последовательность нуклеиновой кислоты, кодирующая альфа-амилазу по любому из пп.1-9.
17. Клетка-хозяин, содержащая последовательность нуклеиновой кислоты по п.16.
18. Способ получения альфа-амилазы по любому из пп.1-9, включающий следующие стадии:
(a) выращивание клетки-хозяина по п.17 в условиях, приводящих к экспрессии гибридной альфа-амилазы; и
(b) извлечение гибридной альфа-амилазы.
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