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DK2507372T3 - Polypeptider med glucoamylaseaktivitet og polynukleotider, der koder for dem - Google Patents

Polypeptider med glucoamylaseaktivitet og polynukleotider, der koder for dem Download PDF

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Publication number
DK2507372T3
DK2507372T3 DK10785298.0T DK10785298T DK2507372T3 DK 2507372 T3 DK2507372 T3 DK 2507372T3 DK 10785298 T DK10785298 T DK 10785298T DK 2507372 T3 DK2507372 T3 DK 2507372T3
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seq
polypeptide
ala
ser
thr
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DK10785298.0T
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Sara Landvik
Marc Dominique Morant
Keiichi Ayabe
Guillermo Coward-Kelly
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Novozymes North America Inc
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    • 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
    • C12P7/14—Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
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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.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
    • C12N9/242—Fungal source
    • 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
    • C12N9/2428—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
    • 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
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/02—Monosaccharides
    • 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
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01001—Alpha-amylase (3.2.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01003—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
    • 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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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (17)

1. Isoleret polypeptid med glucoamylaseaktivitet, der er udvalgt fra gruppen, som består af: (a) et polypeptid, som omfatter en aminosyresekvens, der har mindst 90%, mere fortrinsvis mindst 91%, mere fortrinsvis mindst 92%, endnu mere fortrinsvis mindst 93%, mest fortrinsvis mindst 94% og allermest fortrinsvis mindst 95%, såsom mindst 96%, mindst 97%, mindst 98%, mindst 99% eller endda 100% identitet med aminosyrerne 19 til 573 ifølge SEQ ID NO: 2, SEQ ID NO: 4 eller SEQ ID NO: 6; (b) et polypeptid, som omfatter en aminosyresekvens, der har mindst 90%, mere fortrinsvis mindst 91%, mere fortrinsvis mindst 92%, endnu mere fortrinsvis mindst 93%, mest fortrinsvis mindst 94% og allermest fortrinsvis mindst 95%, såsom mindst 96%, mindst 97%, mindst 98%, mindst 99% eller endda 100% identitet med det katalytiske domæne, som er vist som aminosyrerne 22 til 476 ifølge SEQ ID NO: 2, SEQ ID NO: 4 eller SEQ ID NO: 6; (c) et polypeptid, der kodes for af et polynukleotid omfattende en nukleotidsekvens, som har mindst 90%, mere fortrinsvis mindst 91%, mere fortrinsvis mindst 92%, endnu mere fortrinsvis mindst 93%, mest fortrinsvis mindst 94% og allermest fortrinsvis mindst 95%, såsom endda mindst 96%, 97%, 98%, 99% eller 100% identitet med den kodende sekvens ifølge SEQ ID NO: 1, SEQ ID NO: 3 eller SEQ ID NO: 5, der koder for aminosyrerne 19 til 573 ifølge SEQ ID NO: 2, SEQ ID NO: 4 eller SEQ ID NO: 6.
2. Polypeptid ifølge krav 1, som omfatter eller består af aminosyrerne 19 til 573 ifølge SEQ ID NO: 2, SEQ ID NO: 4 eller SEQ ID NO: 6; eller et fragment deraf med glucoamylaseaktivitet.
3. Polypeptid ifølge krav 1 eller 2, der kodes for af et polynukleotid, som omfatter eller består af nukleotidsekvensen ifølge SEQ ID NO: 1, SEQ ID NO: 3 eller SEQ ID NO: 5; eller en delsekvens deraf, der koder for et fragment med glucoamylaseaktivitet.
4. Polypeptid ifølge et hvilket som helst af kravene 1-3, der kodes for af polynukleotidet, som indgår i plasmid indeholdt i E. coli DSM 23221.
5. Isoleret polynukleotid omfattende en nukleotidsekvens, der koder for polypeptidet ifølge et hvilket som helst af kravene 1-4.
6. Nukleinsyrekonstruktion, som omfatter polynukleotidet ifølge krav 9 operativt forbundet til en eller flere (adskillige) kontrolsekvenser, der styrer fremstillingen af polypeptidet i en ekspressionsvært.
7. Rekombinant ekspressionsvektor, som omfatter nukleinsyrekonstruktionen ifølge krav 6.
8. Rekombinant værtscelle, som omfatter nukleinsyrekonstruktionen ifølge krav 6.
9. Fremgangsmåde til fremstilling af polypeptidet ifølge et hvilket som helst af kravene 1-4, hvilken fremgangsmåde omfatter: (a) dyrkning af en værtscelle, som omfatter en nukleinsyrekonstruktion, hvilken nukleinsyrekonstruktion omfatter en nukleotidsekvens, der koder for polypeptidet, under betingelser, som er befordrende for fremstilling af polypeptidet; og (b) indvinding af polypeptidet.
10. Fremgangsmåde til fremstilling af polypeptidet ifølge et hvilket som helst af kravene 1-4, hvilken fremgangsmåde omfatter: (a) dyrkning af en transgen plante eller en plantecelle, som omfatter et polynukleotid, hvilket polynukleotid koder for polypeptidet, under betingelser, som er befordrende for fremstilling af polypeptidet; og (b) indvinding af polypeptidet.
11. Transgen plante, plantedel eller plantecelle omfattende et polynukleotid, hvilket polynukleotid koder for polypeptidet ifølge et hvilket som helst af kravene 1-4.
12. Anvendelse af et polypeptid ifølge et hvilket som helst af kravene 1-4 til fremstilling af sirup og/eller et fermenteringsprodukt.
13. Anvendelse ifølge krav 12, hvor udgangsmaterialet er gelatineret eller ugelatineret stivelsesholdigt materiale.
14. Anvendelse af et polypeptid ifølge et hvilket som helst af kravene 1-4 til brygning.
15. Sammensætning, som omfatter en alfa-amylase og et polypeptid ifølge et hvilket som helst af kravene 1-4.
16. Fremgangsmåde til fremstilling af et fermenteringsprodukt fra stivelsesholdigt materiale, hvilken fremgangsmåde omfatter følgende trin: (a) at gøre stivelsesholdigt materiale flydende; (b) at forsukre materialet, som er gjort flydende; og (c) at fermentere med en fermenterende organisme; hvor trin (b) udføres ved anvendelse af mindst en glucoamylase ifølge et hvilket som helst af kravene 1-4.
17. Fremgangsmåde til fremstilling af et fermenteringsprodukt fra stivelsesholdigt materiale, hvilken fremgangsmåde omfatter følgende trin: (a) at forsukre stivelsesholdigt materiale ved en temperatur, som er lavere end den laveste gelatineringstemperatur for det stivelsesholdige materiale; og (b) at fermentere med en fermenterende organisme, hvor trin (a) udføres ved anvendelse af mindst en glucoamylase ifølge et hvilket som helst af kravene 1-4.
DK10785298.0T 2009-11-30 2010-11-30 Polypeptider med glucoamylaseaktivitet og polynukleotider, der koder for dem DK2507372T3 (da)

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US26497709P 2009-11-30 2009-11-30
PCT/US2010/058438 WO2011066576A1 (en) 2009-11-30 2010-11-30 Polypeptides having glucoamylase activity and polynucleotides encoding same

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US (4) US8916359B2 (da)
EP (1) EP2507372B1 (da)
CN (1) CN102869771B (da)
AU (1) AU2010276468B2 (da)
CA (1) CA2782154C (da)
DK (1) DK2507372T3 (da)
ES (1) ES2534098T3 (da)
MX (1) MX2012006096A (da)
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US10125380B2 (en) 2018-11-13
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