DK2510093T3 - Proteasevarianter - Google Patents
Proteasevarianter Download PDFInfo
- Publication number
- DK2510093T3 DK2510093T3 DK10803307.7T DK10803307T DK2510093T3 DK 2510093 T3 DK2510093 T3 DK 2510093T3 DK 10803307 T DK10803307 T DK 10803307T DK 2510093 T3 DK2510093 T3 DK 2510093T3
- Authority
- DK
- Denmark
- Prior art keywords
- protease
- polypeptide
- variant
- ala
- protease activity
- Prior art date
Links
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)
- C12N9/58—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- 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
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
-
- 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)
- C12Y304/24—Metalloendopeptidases (3.4.24)
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Claims (14)
1. Fremgangsmåde ti! fremstilling af en proteasevariant med proteaseaktivitet og forbedret termostabilitet i sammenligning med sfamproteasen, hvilken fremgangsmåde omfatter dyrkning af en celle, hvori der er indført en ekspressionsvektor, som omfatter følgende operativt forbundne elementer: føj en transskriptionspromofor, fbj et poiynukleotidmoiekyie, der koderfor en proteasevariani, som udviser mindst SS% identitet medgroteasén Vist i aminosyrerne Ϊ til 177 ifølge SEQID NO: 2, og som omfatter mindst én modifikation i sammenligning med aminosyrerne 1 til 177 ifølge SEQ ID NO: 2 i mindst én position vaigt blandt de følgende: 27,79,82,87,112,142, 2, 5,6, 8,26,41,43,46, 49, 63, 64, 73, 88, 104,114, 115,118,126,152,157,158 og173, hvilket pøtynukleotidmotekyfe er fremstillet ved at indføre mindst én mutation i et DNA-molekyie, der koder for en protease, og (c) en fm nssiy ptiemstermi nater, hvorved cellen udtrykker proteasevarianten, der kodes for af polynukleoisdmolekylet; og indvinding af proteasevarianten.
2. Fremgangsmåde ifølge krav 1, hvor proteasevarianten, der kodes for af poHmuk eofidmolekylet, omfatter mindst én modifikation valgt fra gruppen bestående af A27K, A27G, A27V, Q53K, Q53R, T54R, D79K, D79L D79M, Y82F, S87P, S87G. A112P, D142L R2P, AS5, C6R, AGS, N26R, S41R, Y43F, T46R, S49R, A73C. P81R, N88R, D104R, D104P, T114P, S115R, T118V, T124L, T124V, A126V, M152R, S157K, Q158Wog M73V.
3. Fremgangsmåde ifølge et hvilket som helst af kravene 1 eller 2. hvor proteasevarianten, dér: kodes for af poiynuk|eotidmd!éky|et5 drnfader m én af følgende kombinationer af modifikationer: AS5/D7S^S8lRi Ii5/D79L/S37P/A112P/D142L. AS6/N26R/D79L/S87P/A112P/D142L, C6R/D79L/S87P, AG8/D79L./S87P. N26R/D79L/S87P, N28R/r46R/D79L/S87P/A1 12P/D142L, A27G/D79L/S87P/A112P/D142L, M7K/D7^ 12P/D142L, A27K/D73L/S87R/A112P/T124V7D142L, A27V/079IJG87P/A112P/D142U S41R/D79L/S87P, S41R/Q79L/S87P/A112P/D142L, S41R/ppUS87PfA112P/D142L/S157K, Y43F/D79L/S87P/A112P/D142L, T48R/D79L/S87P, Y48Rd379L/S87P/A112P/D142L, T46R/D79L/S87P/T116V/D142L, S49R/D79L/S87P, Q53l®79L/S87P/i 173Y, G53R/D79L/S87P, 1fS41^p79y867P, D79L/S87P/A112P, D79hP81Ra87P/A112P/pi42t, D79L/S87P, D79L/S87P/A112P, D79lp87PiAi:12P7D142L, D79L/S87P/A112P/D142L/S157K, ; D79L/S87P/A412P/T 1 24l/D1 42L, P79L/S87P/A1 1 2P/T124Y«28WD 142 L, mmsm>w 12P7T1 24\//D142L. D79L/S87P/D104R, D79L/S87P/D142L, rørø87R/(488Rs D79L/S87P/Q158W, P79t,/S87P/S11$R( D79US87P/S157K, TOi^S87P7ri 14P, P79ySl7P/T118V, D79L/Y82F/SS7P/A112P/T124V/D142L, D79L/Y82F/S87P/A112P/T124V/P142L, A27K/D79L/Y82F/S87G/D104P/A112P/A128¥/D142L, A27K/Y82F/S87G/D1Q4P/A112P/A126V/D142L. A27K/D79L/Y82F/D104P/A112P/A126V/D142L, A27K/Y82F/0! 04P/A112P/A126V/D142L.
4. Fremgangsmåde ifølge krav 1, hvor proteasevarianfen omfatter mindst Ih higdiffkétipn i mindst én position valgt fra gruppen bestående afpositionerne: 27, 79, 82, 87, 1Q4A112,126 og 142.
5. Fremgangsmåde ifølge krav 4, hvor proteasevarianten omfatter mindst én af modifikationerne valgt fra gruppen bestående af: A27K, D79L, Y82F, S87G, S87P, D1Q4P, A112P, A126V og D142L. 8. iPeiypeptid med proteaseaktivitet, som udviser mindst 85% kfehtitet med proteasen vist tiamihosyferne 1 til 177 ifølge SEQ ID NG: 2, og som omfatter mindst én modifikation i sammenligning med aminosyreme 1 til 177 ifølge SEQ ID NO. 2 i mindst én position våigt blandt de følgende: 27, 79, 82, 87,112, 142. 2, 5, 6, 8, 26, 41,43,46, 49, 53, 54, 73,88,104, 144, 415 116, 126, 152, 157, 158 og 173, hvor poiypeptidet har forbedret termostabilitet i i«^meRiigning:med'proteas!!^''Yiétéi8mthosyrerhe 1 til 177 ifølge SEQ ID NO: 2.
7. Pøiypeptid med proteaseaktivitet ifølge krav 6, som omfatter mindst én modifikation valgt blandt de følgende: A27K, A27G, A27V, Q53K, G53R, T54R, D79K. D79L, D79M, Y82F, S87P, S87G, A112P, D142L, R2P, Δ85, C6R, Δ08, N26R, S41R. Y43F, T46R. S49R, A73C, P81R, N88R, D104R, D104P, T114P, S115R, T116V, T124U, T124V, A126V, M152R, S157K, Q158W og I173V. 8: Pøiypeptid med proteaseaktivitet iføige krav 6, hlpr poiypeptidet omfattet rhindst én modifikation i mindst én position valgt fra gruppen bestående af posttfonerne: 27, 79, 82, 87, 104, 112, 126 og 142,
9. Poiypeptid med proteaseaktivitet iføige krav 8, hvor poiypeptidet omfatter mindst én af rhddifikatiohérhé bestående af: A27R, D79L, Y82F, S87G, S87P, D4G4P,
10, Pdiyp^gifcf ifhéd proteaseaktivitet ifølge ét hvilket som helst af kravene 6-9, hvilket poiypeptib omfatter et sæt af modifikationer valgt blandt de følgende; AS5/Q79US87P, AS5/D79L/S87P/A112P/D142L &S5/N2SR/D79L/S87P/A112P/D1421, C6R/P791/S87P, &G8/079yS87P, M26R/D791/S87P, N26R/T46R/079I/S87P/A112P/P142L, A27G/D7917S87P/A112P/0142LA27K/D79L/S87P/A112P/D142L, A27K/D79t,7S87p/Ai 12P/T124V/D142L, A27V/D79US87P/A112P/D142L, S41R/D79I/S87P, S41R/D79US87P/A112P/D1421, S41R/D79L/S87 P/A112P/D142L/S157K, Y43F/D7SL/S87FÉM 12P/D142L. T46R/D79US87P, T46R/D79L/S87P/A112P/D142L, T46R'D79L/S87P/T 116V/D142L, S49R/D79L/S87 P, Q53K/D79L/S 87P/1173V. G53R/D79L/S87P, T54R/D79L/S87P, D79U S87P/A112P, D79UP81R/S87P/A112P/D142L, Q7917S87F, D79k/i87P/Ai 12P, D79i../S87FfA112P/D142L, 079L/S87P/A112PVQ142kmS7K, D79L/S87P/A112P/T124L/D142L, D79US87P/A 112P/T124V/A126V/D142L, D79L/S87P/A112P/T124V/D142L. D79US87P/D104R, D79L/S87P/D142L D79L/S87P/N88R, D79L/S87P/G158W. D79L/S87P/S115R D79L/S87P/S157K, D79L/S87P/T114P, D79L/S87P/T116V/, P79L/^82R/S87P/A112P/T124V/0102L 079t/Y82Fi^ 24W0142L A27K/D79UY82F/S87G/D104P/A112P/A126V/D1421, A27K/Y82P/S87G/D104P/A112P/A126V/D142L, A2 7 K7 D7 9L/Y82F/D104 P/A112P/A126V/D142L, A27K/Y82F/D104P/A112P/A126V/D142L. 11 løøifø^r^NkiSid^resekvenSi; som; omfatter en nukleinsyresekvens, Jef koder for poiypeptidet sned protea.seaktivitet ifsige et hvilket som helst af kravene 6-10.
12, Nukieinsyrekonstruktionr som omfatter nuklemsyresekvensen følge krav 11. der er operativt forbundet tji en eller flere kontrolsekvenser, som styret frem^ltlllngéh af proteasen i en egnet ekspressionsvært.
13. Rekombinant ekspressionsvektor, som omfatter nukleinsyrekonstruktionen ifølge krav 12,
14. Rékomøihsht værtscelle, som omfatter nukleinsyrekonstruktionen ifølge krav 12 og/e!!er ekspressiGnsvektoren tfølge krav 13.
15, Fremgangsmåde til fremstilling af poiypeptidet røéd proteaseaktivitet ifølge et hvilket som helst af kravene 6-10, hvilken fremgangsmåde omfatter: (a) dyrkning af vasøseSIføb ifølge kfp 14: for at fremstille en supernatant, som omfatter polypeptidet med proteaseaktivitet; og (b) indvinding af polypeptidet med proteaseaktivitet. li, Tfipogen plante ellerplantedel, som kan udtrykkes! poiypeptld med proteaseaktivitet ifølge oi P^iikét sorr? helst af kravene 6Ί0. 17. fremgangsmidé fffremstilling åf et ferroenteringsprodukt, hvilken fremgangsmåde omfatter (a) fermentering ved anvendelse af en fermenterende mikroorganisme og et inærværelse af et polypeptid med proteaseaktivitet ifølge et hvilket som helst af kravene 6-10 og {fe} fremstilling af fermenteringsproduktet ud fra det fermenterede carbohydratholdige materiale.
18. Fremgangsmåde ifølge krav l 1, hvor fermenteringsproduktet er ethanol og/eilertørret bærm e idlstlSlers dri ed gralns - O DG).
19. Anvendelse af mindst ét polypeptid med proteaseaktivitetifølge et hvilket som helst af kravene 6-10 ved en fremgangsmåde til fremstilling af et fermenieringsprodukt fortrinsvis ethano!.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US28560109P | 2009-12-11 | 2009-12-11 | |
PCT/US2010/059814 WO2011072191A2 (en) | 2009-12-11 | 2010-12-10 | Protease variants |
Publications (1)
Publication Number | Publication Date |
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DK2510093T3 true DK2510093T3 (da) | 2018-06-06 |
Family
ID=44146191
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DK10803307.7T DK2510093T3 (da) | 2009-12-11 | 2010-12-10 | Proteasevarianter |
Country Status (9)
Country | Link |
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US (2) | US9040280B2 (da) |
EP (1) | EP2510093B1 (da) |
CN (2) | CN102753680B (da) |
AU (1) | AU2010328033B2 (da) |
CA (1) | CA2783820C (da) |
DK (1) | DK2510093T3 (da) |
ES (1) | ES2668202T3 (da) |
MX (2) | MX2012006548A (da) |
WO (1) | WO2011072191A2 (da) |
Families Citing this family (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012088303A2 (en) | 2010-12-22 | 2012-06-28 | Novozymes North America, Inc. | Processes for producing fermentation products |
EA201490568A1 (ru) | 2011-09-06 | 2014-06-30 | Новозимс А/С | Варианты глюкоамилазы и кодирующие их полинуклеотиды |
IN2014CN03467A (da) | 2011-10-11 | 2015-10-16 | Novozymes North America Inc | |
ES2638669T3 (es) | 2011-10-11 | 2017-10-23 | Novozymes A/S | Variantes de glucoamilasa y polinucleótidos que las codifican |
EP2785847B1 (en) * | 2011-12-02 | 2017-07-26 | Novozymes A/S | Liquefaction process with selected alpha-amylases and proteases |
US9175316B2 (en) * | 2012-12-12 | 2015-11-03 | Ebio, Llc | Efficient production of biofuels from cells carrying a metabolic-bypass gene cassette |
ES2935920T3 (es) | 2012-03-30 | 2023-03-13 | Novozymes North America Inc | Procesos de elaboración de productos de fermentación |
EP4209595A1 (en) | 2012-03-30 | 2023-07-12 | Novozymes North America, Inc. | A method of dewatering whole stillage |
PL3013967T3 (pl) | 2013-06-24 | 2022-03-21 | Novozymes A/S | Sposoby odzyskiwania oleju z procesów wytwarzania produktów fermentacji i sposoby wytwarzania produktów fermentacji |
US11939552B2 (en) | 2013-06-24 | 2024-03-26 | Novozymes A/S | Process of recovering oil |
HUE040545T2 (hu) | 2013-07-17 | 2019-03-28 | Novozymes As | Pullulanáz kimérák és az azokat kódoló polinukleotidok |
EP3063282B1 (en) * | 2013-09-11 | 2020-03-25 | Novozymes A/S | Processes for producing fermentation products |
CA2932239C (en) | 2014-01-22 | 2023-03-14 | Novozymes A/S | Pullulanase variants and polynucleotides encoding same |
AR102419A1 (es) | 2014-10-23 | 2017-03-01 | Novozymes As | Variantes de glucoamilasa y polinucleótidos que las codifican |
EP3487995B1 (en) | 2016-07-21 | 2021-05-26 | Novozymes A/S | Serine protease variants and polynucleotides encoding same |
US10889836B2 (en) | 2016-11-23 | 2021-01-12 | Novozymes A/S | Yeast for ethanol production |
EP3609631A1 (en) * | 2017-04-14 | 2020-02-19 | Novozymes A/S | Processes for solubilizing municipal solid waste with enzyme compositions comprising protease and enzyme compositions thereof |
CN107384900B (zh) * | 2017-08-01 | 2019-08-27 | 中国农业科学院饲料研究所 | 一种真菌来源的酸性蛋白酶6749及其基因和应用 |
WO2019161227A1 (en) | 2018-02-15 | 2019-08-22 | Novozymes A/S | Improved yeast for ethanol production |
BR112021017085A2 (pt) | 2019-04-02 | 2022-02-08 | Novozymes As | Processo de produção de um produto de fermentação |
CN113544226A (zh) * | 2019-04-08 | 2021-10-22 | 诺维信公司 | 提取明胶的方法 |
CN112048518B (zh) * | 2020-08-22 | 2022-06-24 | 江西农业大学 | 一种玉米醇溶蛋白降解酶的制备方法及其应用 |
CN114574468A (zh) * | 2022-02-17 | 2022-06-03 | 皖西学院 | 一种高活性高热稳定性的蛋白酶突变体及其制备、应用 |
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DK122686D0 (da) | 1986-03-17 | 1986-03-17 | Novo Industri As | Fremstilling af proteiner |
PT97110B (pt) | 1990-03-23 | 1998-11-30 | Gist Brocades Nv | Processo para catalisar reaccoes acelaraveis por enzimas, mediante adicao ao meio reaccional de sementes de plantas transgenicas e para obtencao das referidas sementes |
US5231017A (en) | 1991-05-17 | 1993-07-27 | Solvay Enzymes, Inc. | Process for producing ethanol |
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US5689054A (en) | 1994-03-17 | 1997-11-18 | The United States Of America As Represented By The Secretary Of Agriculture | Low phytic acid mutants and selection thereof |
DE4422198C2 (de) | 1994-06-24 | 1997-08-28 | Audi Ag | Verfahren zum Steuern der elektrischen Beheizung eines Katalysators |
AU2705895A (en) | 1994-06-30 | 1996-01-25 | Novo Nordisk Biotech, Inc. | Non-toxic, non-toxigenic, non-pathogenic fusarium expression system and promoters and terminators for use therein |
NZ330940A (en) | 1997-07-24 | 2000-02-28 | F | Production of consensus phytases from fungal origin using computer programmes |
JP4988124B2 (ja) | 2000-06-02 | 2012-08-01 | ノボザイムス アクティーゼルスカブ | クチナーゼ変異体 |
BR0214653B1 (pt) * | 2001-12-07 | 2015-01-06 | Novozymes As | Construção de ácido nucléico, vetor de expressão recombinante, célula de microorganismo hospedeiro recombinante, método para produzir um polipeptídeo, uso de pelo menos uma protease, método para melhorar o valor nutricional de uma ração animal, aditivo para ração animal, composição de ração animal, e, método para o tratamento de proteínas egetais. |
CN101087809B (zh) * | 2004-12-22 | 2015-01-21 | 诺维信公司 | 杂合酶 |
DK2390321T3 (da) * | 2005-10-12 | 2015-02-23 | Procter & Gamble | Anvendelse og fremstilling af en opbevaringsstabil neutral metalloprotease |
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2010
- 2010-12-10 CN CN201080063729.3A patent/CN102753680B/zh active Active
- 2010-12-10 US US13/510,076 patent/US9040280B2/en active Active
- 2010-12-10 DK DK10803307.7T patent/DK2510093T3/da active
- 2010-12-10 MX MX2012006548A patent/MX2012006548A/es active IP Right Grant
- 2010-12-10 ES ES10803307.7T patent/ES2668202T3/es active Active
- 2010-12-10 MX MX2015008451A patent/MX355362B/es unknown
- 2010-12-10 EP EP10803307.7A patent/EP2510093B1/en active Active
- 2010-12-10 CA CA2783820A patent/CA2783820C/en active Active
- 2010-12-10 AU AU2010328033A patent/AU2010328033B2/en active Active
- 2010-12-10 CN CN201510178227.XA patent/CN104774823B/zh active Active
- 2010-12-10 WO PCT/US2010/059814 patent/WO2011072191A2/en active Application Filing
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2015
- 2015-04-20 US US14/690,937 patent/US10138473B2/en active Active
Also Published As
Publication number | Publication date |
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WO2011072191A3 (en) | 2012-04-19 |
US20150218545A1 (en) | 2015-08-06 |
CN102753680B (zh) | 2015-05-13 |
AU2010328033B2 (en) | 2014-12-04 |
CN104774823A (zh) | 2015-07-15 |
EP2510093B1 (en) | 2018-02-21 |
CA2783820A1 (en) | 2011-06-16 |
EP2510093A2 (en) | 2012-10-17 |
US10138473B2 (en) | 2018-11-27 |
CN104774823B (zh) | 2020-04-10 |
CN102753680A (zh) | 2012-10-24 |
US20120309067A1 (en) | 2012-12-06 |
CA2783820C (en) | 2018-10-23 |
US9040280B2 (en) | 2015-05-26 |
AU2010328033A1 (en) | 2012-06-07 |
WO2011072191A2 (en) | 2011-06-16 |
ES2668202T3 (es) | 2018-05-17 |
MX355362B (es) | 2018-04-17 |
MX2012006548A (es) | 2012-07-10 |
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