DK2588604T3 - Polypeptider med beta-glucosidaseaktivitet og polynukleotider, der koder for dem - Google Patents

Polypeptider med beta-glucosidaseaktivitet og polynukleotider, der koder for dem Download PDF

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Publication number
DK2588604T3
DK2588604T3 DK11736227.7T DK11736227T DK2588604T3 DK 2588604 T3 DK2588604 T3 DK 2588604T3 DK 11736227 T DK11736227 T DK 11736227T DK 2588604 T3 DK2588604 T3 DK 2588604T3
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polypeptide
beta
cellulosic material
cell
acid
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DK11736227.7T
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English (en)
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Marc Morant
Shamkant Patkar
Hanshu Ding
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Novozymes Inc
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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
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • 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
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2445—Beta-glucosidase (3.2.1.21)
    • 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)
    • C12Y302/01021—Beta-glucosidase (3.2.1.21)
    • 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
    • Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (18)

1. Isoleret polypeptid med beta-glucosidaseaktivitet, som er valgt fra gruppen bestående af: (a) et polypeptid, som udviser mindst 95%, f.eks. mindst 96%, mindst 97%, mindst 98%, mindst 99% og 100% sekvensidentitet med det modne polypeptid ifølge SEQ ID NO: 2; og (b) en variant af det modne polypeptid ifølge SEQ ID NO: 2, som omfatter en substitution, deletion og/eller insertion i en eller flere positioner, hvor varianten udviser mindst 85%, f.eks. mindst 86%, mindst 87%, mindst 88%, mindst 89%, mindst 90%, mindst 91%, mindst 92%, mindst 93%, mindst 94%, mindst 95%, mindst 96%, mindst 97%, mindst 98%, mindst 99% sekvensidentitet med det modne polypeptid ifølge SEQ ID NO: 2, og hvor varianten har forbedret termisk stabilitet i forhold til det modne polypeptid ifølge SEQ ID NO: 2.
2. Polypeptid ifølge krav 1, hvilket polypeptid omfatter eller består af SEQ ID NO: 2 eller det modne polypeptid deraf.
3. Polypeptid ifølge krav 1 eller 2, hvilket polypeptid kodes for af polynukleotidet indeholdt i plasmid pUC19 D55EX, hvilket plasmid er indeholdt i E. coli NRRL B-50395.
4. Sammensætning, som omfatter polypeptidet ifølge et hvilket som helst af kravene 1 -3.
5. Isoleret polynukleotid, som koder for polypeptidet ifølge et hvilket som helst af kravene 1-3.
6. Fremgangsmåde til fremstilling af polypeptidet ifølge et hvilket som helst af kravene 1-3, hvilken fremgangsmåde omfatter: (a) dyrkning af en celle, som i sin vildtypeform fremstiller polypeptidet, under betingelser, der er befordrende for fremstilling af polypeptidet; og (b) indvinding af polypeptidet.
7. Fremgangsmåde til fremstilling af et polypeptid med beta-glucosidaseaktivitet, hvilken fremgangsmåde omfatter: (a) dyrkning af en værtscelle, som omfatter polynukleotidet ifølge krav 5, under betingelser, der er befordrende for fremstilling af polypeptidet; og (b) indvinding af polypeptidet.
8. Transgen plante, plantedel eller plantecelle, som er transformeret med et polynukleotid, hvilket polynukleotid koder for polypeptidet ifølge et hvilket som helst af kravene 1-3.
9. Fremgangsmåde til fremstilling af polypeptidet med beta-glucosidaseaktivitet, hvilken fremgangsmåde omfatter: (a) dyrkning af den transgene plante eller plantecelle ifølge krav 8 under betingelser, som er befordrende for fremstilling af polypeptidet; og (b) indvinding af polypeptidet.
10. Fremgangsmåde til fremstilling af en mutant af en stamcelle, hvilken fremgangsmåde omfatter inaktivering af et polynukleotid, der koder for polypeptidet ifølge et hvilket som helst af kravene 1-3, hvilket resulterer i, at mutanten fremstiller mindre af polypeptidet end stamcellen.
11. Dobbeltstrenget hæmmende RNA (dsRNA)-molekyle, som omfatter en undersekvens af polynukleotidet ifølge krav 5, hvor dsRNA'et eventuelt er et siRNA- eller miRNA-molekyle.
12. Fremgangsmåde til hæmning af ekspressionen af et polypeptid med beta-glucosidaseaktivitet i en celle, hvilken fremgangsmåde omfatter administration til cellen eller ekspression i cellen af det dobbeltstrengede RNA (dsRNA)-molekyle ifølge krav 11.
13. Fremgangsmåde til nedbrydning eller omdannelse af et celluloseholdigt materiale, hvilken fremgangsmåde omfatter: behandling af det celluloseholdige materiale med en enzymsammensætning i nærværelse af polypeptidet med beta-glucosidaseaktivitet ifølge et hvilket som helst af kravene 1-3.
14. Fremgangsmåde ifølge krav 13, hvilken fremgangsmåde endvidere omfatter indvinding af det nedbrudte celluloseholdige materiale.
15. Fremgangsmåde til fremstilling af et fermenteringsprodukt, hvilken fremgangsmåde omfatter: (a) forsukring af et celluloseholdigt materiale med en enzymsammensætning i nærværelse af polypeptidet med beta-glucosidaseaktivitet ifølge et hvilket som helst af kravene 1-3; (b) fermentering af det forsukrede celluloseholdige materiale med en eller flere fermenterende mikroorganismer for at fremstille fermenteringsproduktet; og (c) indvinding af fermenteringsproduktet fra fermenteringen.
16. Fremgangsmåde til fermentering af et celluloseholdigt materiale, hvilken fremgangsmåde omfatter: fermentering af det celluloseholdige materiale med en eller flere fermenterende mikroorganismer, hvor det celluloseholdige materiale er forsukret med en enzymsammensætning i nærværelse af et polypeptid med beta-glucosidaseaktivitet ifølge et hvilket som helst af kravene 1-3.
17. Fremgangsmåde ifølge krav 16, hvor fermenteringen af det celluloseholdige materiale frembringer et fermenteringsprodukt.
18. Fremgangsmåde ifølge krav 17, hvilken fremgangsmåde endvidere omfatter indvinding af fermenteringsproduktet fra fermenteringen.
DK11736227.7T 2010-06-30 2011-06-30 Polypeptider med beta-glucosidaseaktivitet og polynukleotider, der koder for dem DK2588604T3 (da)

Applications Claiming Priority (2)

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US36035110P 2010-06-30 2010-06-30
PCT/US2011/042682 WO2012003379A1 (en) 2010-06-30 2011-06-30 Polypeptides having beta-glucosidase activity and polynucleotides encoding same

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US (1) US8581042B2 (da)
EP (1) EP2588604B1 (da)
CN (1) CN103108951A (da)
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WO (1) WO2012003379A1 (da)

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