DK2841567T3 - GH61-polypeptidvarianter og polynukleotider, som koder for dem - Google Patents

GH61-polypeptidvarianter og polynukleotider, som koder for dem Download PDF

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DK2841567T3
DK2841567T3 DK13720735.3T DK13720735T DK2841567T3 DK 2841567 T3 DK2841567 T3 DK 2841567T3 DK 13720735 T DK13720735 T DK 13720735T DK 2841567 T3 DK2841567 T3 DK 2841567T3
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variant
polypeptide
another aspect
parent
thermostability
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Janine Lin
Doreen Bohan
Michelle Maranta
Matt Sweeney
Mark Wogulis
Elizabeth Znameroski
Frank Winther Rasmussen
Leslie Beresford
Michael Lamsa
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Novozymes Inc
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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
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • 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
    • C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
    • 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
    • C11D3/38654—Preparations containing enzymes, e.g. protease or amylase containing oxidase or reductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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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
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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    • 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
    • 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
    • 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)
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
    • D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
    • D21C5/005—Treatment of cellulose-containing material with microorganisms or enzymes
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21H—PULP COMPOSITIONS; PREPARATION THEREOF NOT COVERED BY SUBCLASSES D21C OR D21D; IMPREGNATING OR COATING OF PAPER; TREATMENT OF FINISHED PAPER NOT COVERED BY CLASS B31 OR SUBCLASS D21G; PAPER NOT OTHERWISE PROVIDED FOR
    • D21H17/00—Non-fibrous material added to the pulp, characterised by its constitution; Paper-impregnating material characterised by its constitution
    • D21H17/005—Microorganisms or enzymes
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    • C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
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    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • 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)
    • C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

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  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
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  • Oil, Petroleum & Natural Gas (AREA)
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  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Claims (16)

1. GH61-polypeptidvariant, som omfatter en Q138C,G,K,L- eller M-substitution i en position svarende til position 138 i stampolypeptidet ifølge SEQ ID NO: 30, hvor varianten har cellulolytisk forbedrende aktivitet; mindst 80%, mindst 81%, mindst 82%, mindst 83%, mindst 84%, mindst 85%, mindst 86%, mindst 87%, mindst 88%, mindst 89%, mindst 90%, mindst 95%, mindst 96%, mindst 97%, mindst 98% eller mindst 99%, men mindre end 100%, sekvensidentitet med aminosyresekvensen for stam-GH61-polypeptidet ifølge SEQ ID NO: 30; og - forøget termostabilitet i sammenligning med stampolypeptidet ifølge SEQ ID NO: 30.
2. GH61-polypeptidvariant, som omfatter en Q138E-substitution i en position svarende til position 138 i stampolypeptidet ifølge SEQ ID NO: 30, hvor varianten har: - cellulolytisk forbedrende aktivitet; mindst 97%, mindst 98%, mindst 99%, men mindre end 100%, sekvensidentitet med aminosyresekvensen for stam-GH61-polypeptidet ifølge SEQ ID NO: 30; og forøget termostabilitet i sammenligning med stampolypeptidet ifølge SEQ ID NO: 30.
3. Variant ifølge krav 1 eller 2, som endvidere omfatter en eller flere substitutioner valgt fra gruppen bestående af S26I; G32E,S; Y34F; V40A; N41T; Q42I,E,V; S47E,L,R; S56C,E,T; S72Q,T; T102KP; A123R; V149I; D152S; T163E,F,V; V164C,L; I166L; S169R.C; S186F,K,T,Y; F200I,V; G207P; S213E; S219E,M,Q,C; K222R; S234G.K; A246P; N249Q,R,C og A250C.
4. Variant ifølge et hvilket som helst af kravene 1-3, som endvidere omfatter en substitution i en eller flere positioner svarende til positionerne 111, 152, 155, 162, 96, 98, 200, 202, 204, 105, 154,188, 189, 216 og 229 i det modne polypeptid ifølge SEQ ID NO: 30, hvor varianten har cellulolytisk forbedrende aktivitet.
5. Variant ifølge krav 4, som omfatter en eller flere substitutioner valgt fra gruppen bestående af L111V, D152S, M155L, A162W, I96V, F98L, F200I, I202L, I204V, E105P,K; E154L; G188A,W; N189K; A216L,Y og A229W,H,I,Y.
6. Isoleret polynukleotid, som koder for varianten ifølge et hvilket som helst af kravene 1-5.
7. Rekombinant værtscelle, som omfatter polynukleotidet ifølge krav 6.
8. Fremgangsmåde til fremstilling af en GH61-polypeptidvariant, hvilken fremgangsmåde omfatter: dyrkning af den rekombinante værtscelle ifølge krav 7 under betingelser, som er egnede til ekspression af varianten.
9. Transgen plante, plantedel eller plantecelle, som er transformeret med polynukleotidet ifølge krav 6.
10. Fremgangsmåde til fremstilling af varianten ifølge et hvilket som helst af kravene 1-5, hvilken fremgangsmåde omfatter: dyrkning af en transgen plante eller en plantecelle, som omfatter et polynukleotid, der koder for varianten, under betingelser, som er befordrende for fremstilling af varianten.
11. Fremgangsmåde til nedbrydning eller omdannelse af et celluloseholdigt materiale, hvilken fremgangsmåde omfatter: behandling af det celluloseholdige materiale med en enzymsammensætning i nærværelse af GH61-polypeptidvarianten med cellulolytisk forbedrende aktivitet ifølge et hvilket som helst af kravene 1-5.
12. Fremgangsmåde til fremstilling af et fermenteringsprodukt, hvilken fremgangsmåde omfatter: (a) forsukring af et celluloseholdigt materiale med en enzymsammensætning i nærværelse af GH61-polypeptidvarianten med cellulolytisk forbedrende aktivitet ifølge et hvilket som helst af kravene 1-5; (b) fermentering af det forsukrede celluloseholdige materiale med en eller flere fermenterende mikroorganismer for at fremstille fermenteringsproduktet; og (c) indvinding af fermenteringsproduktet fra fermenteringen.
13. 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 GH61-polypeptidvarianten med cellulolytisk forbedrende aktivitet ifølge et hvilket som helst af kravene 1-5.
14. Sammensætning, som omfatter varianten ifølge et hvilket som helst af kravene 1-5.
15. Totalmedieformulering eller celledyrkningssammensætning, som omfatter varianten ifølge et hvilket som helst af kravene 1-5.
16. Detergentsammensætning, som omfatter et overfladeaktivt middel og varianten ifølge et hvilket som helst af kravene 1-5.
DK13720735.3T 2012-04-27 2013-04-26 GH61-polypeptidvarianter og polynukleotider, som koder for dem DK2841567T3 (da)

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PCT/US2013/038477 WO2013163590A2 (en) 2012-04-27 2013-04-26 Gh61 polypeptide variants and polynucleotides encoding same

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CN (2) CN104245926B (da)
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