FI121925B - Förbättrad sackarifikation av cellulosa genom kloning och amplifikering av b-glukosidasgenen av Trichoderma reesei - Google Patents

Förbättrad sackarifikation av cellulosa genom kloning och amplifikering av b-glukosidasgenen av Trichoderma reesei Download PDF

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FI121925B
FI121925B FI932633A FI932633A FI121925B FI 121925 B FI121925 B FI 121925B FI 932633 A FI932633 A FI 932633A FI 932633 A FI932633 A FI 932633A FI 121925 B FI121925 B FI 121925B
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glucosidase
gene
mold
transformants
bglii
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FI932633A
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Finnish (fi)
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FI932633A0 (fi
FI932633L (fi
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Christopher C Barnett
Timothy Fowler
Sharon Shoemaker
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Genencor Int
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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
    • C11D3/38645—Preparations containing enzymes, e.g. protease or amylase containing cellulase
    • 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
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungi isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G1/00—Preparation of wine or sparkling wine
    • C12G1/02—Preparation of must from grapes; Must treatment and fermentation
    • 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
    • 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
    • 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
    • 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)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/885—Trichoderma
    • 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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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Mycology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Virology (AREA)
  • Botany (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)
  • Detergent Compositions (AREA)
  • Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
  • Seasonings (AREA)

Claims (24)

1. Förfarande för modifiering av expression av ex-tracellulärt β-glukosidas i f ilamentmögel, känneteck- 5 nat av att nämnda filamentmögel transformeras med en ex-pressionsvektor, som innehäller en DNA-sekvens av möglet, vilken sekvens: förmär förbättra expressionen av extracellulärt β-glukosidas för att den innehäller ätminstone en över-10 loppskopia av möglets β-glukosidasgen, varvid filamentmög-let är Trichoderma reesei och nämnda extracellulära β-glukosidasgen av möglet är en bgll-gen som härstammar frän Trichoderma reesei.
2. Förfarande för att förbättra expressionen av 15 extracellulärt β-glukosidas enligt patentkrav 1, känne- tecknat av att nämnda expressionsvektor innehäller hela mögel^-glukosidasgenens kodande omräde och sekven-ser, vilka behövs för transkription och translation av β-glukosidasgenen. 20
3. Förfarande enligt patentkrav 1 eller 2, kän- netecknat av att nämnda f ilamentmögel väljs frän Trichoderma-, Aspergillus-, Neurospora-, Humicola- och Pe-nicillium-släktena.
4. Förfarande enligt patentkrav 3, känneteck- 25 nat av att nämnda filamentmögel är Trichoderma reesei, Trichoderma viridae, Trichoderma koningii, Aspergillus ni- ^ ger, Aspergillus nidulans, Aspergillus wentii, Aspergillus o oryzae, Aspergillus phoenics, Neurospora erässä, Humicola I - - - - o grisea, Penicillum pinophilum eller Penicillum oxalium. £3 30
5. Förfarande enligt nägot av de föregäende päin tentkraven, kännetecknat av att nämnda β-glukosidas- Q_ gen är en bgll-gen, som har erhällits frän Trichoderma co g reesei. g
6. Förfarande enligt patentkrav 5, känneteck- 35 nat av att nämnda bgll-gen omfattar aminosyror, för vilka kodar en nukleotidsekvens 311 - 2 679 enligt figur 1.
7. Förfarande enligt nägot av de föregäende pa-tentkraven, kännetecknat av att det dessutom inne-häller ett steg, i vilket isoleras transformanter med mo-difierad expression av β-glukosidas. 5
8. Förfarande enligt patentkrav 7, känneteck nat av att det dessutom innehäller steg, i vilka (a) odlas nämnda transformanter i förhällanden som tilläter nämnda transformanters tillväxt; och (b) isoleras en rekombinant mögelcellulaskomposit-10 ion producerad av nämnda transformanter.
9. Förfarande enligt patentkrav 8, kännetecknat av att nämnda rekombinanta mögelcellulaskomposition isoleras pä följande sätt: (a) centrifugeras nämnda odlingssubstrat, som in-15 nehäller nämnda transformanter för att bilda en supernatant och pellet, och (b) filtreras nämnda supernatant för att erhälla den rekombinanta mögelcellulaskompositionen.
10. Förfarande enligt patentkrav 9, känneteck-20 nat av att ett antimikrobiellt ämne tillsätts i nämnda rekombinanta mögelcellulaskomposition efter filtreringen.
11. Förfarande enligt nägot av patentkraven 8-10, kännetecknat av att det dessutom innehäller ett steg, väri en expressionsprodukt renas frän nämnda isole- 25 rade rekombinanta mögelcellulaskomposition.
12. Mögelcellulaskomposition, kännetecknad av att den har erhällits frän filamentmögel, som omfattar o ^ rekombinant modifierat β-glukosidas, som har producerats i o med ett förfarande enligt nägot av patentkraven 8-10. £5 30
13. Förfarande för att producera glukos frän cel- x lulosa eller heteroglykaner, kännetecknat av att man Q_ använder en rekombinant mögelcellulaskomposition produce-oo g rad med förfarandet enligt nägot av patentkraven 8 - 10, co vars β-glukosidasaktivitet har förbättrats. CD r ^
14. Förfarande för nedbrytning av cellulosamateri-al som ingär i foder, biomassa eller slam, känneteck-nat av att man använder en rekombinant mögelcellulaskom-position producerad med förfarandet enligt nägot av pa- 5 tentkraven 8 - 10, vars β-glukosidasaktivitet har förbätt-rats.
15. Förfarande för nedbrytning av cellulosa, kännetecknat av att man använder en rekombinant mö-gelcellulaskomposition producerad med förfarandet enligt 10 nägot av patentkraven 8 - 10, vars β-glukosidasaktivitet har förbättrats.
16. Användning av en rekombinant mögelcellulaskom-position producerad med förfarandet enligt nägot av patentkraven 8 - 10, vars β-glukosidasaktivitet har förbätt- 15 rats eller av renat β-glukosidas producerat med förfarandet enligt patentkrav 11 i livsmedelstillsatsämnen för att förbättra livsmedlens smak och arom.
17. Detergent komposition, kännetecknad av att den innehäller en renande effektiv mängd av ett ytak- 20 tivt ämne och ätminstone 0,0001 viktprocent av en rekombinant mögelcellulaskomposition producerad med förfarandet enligt nägot av patentkraven 8 - 10, vars β-glukosidasaktivitet har förbättrats.
18. Transformanter, kännetecknade av att de 25 har förverkligats med förfarandet enligt patentkrav 7.
19. Förfarande enligt patentkrav 13, varvid nämnda ^ cellulaskomposition eller transformanter enligt patentkrav o ^ 18 tillsätts i biomassa för att bilda en blandning för o fermentering av etanol. c3 30
20. Förfarande enligt patentkrav 19, känne- g tecknat av att vid f ermenteringen används jästen CL clausenii, S. cerevisiae, Cellulolyticus acidothermophi- CO to lium, C. brassicae, C. lustinaniae, S. uvarum eller Schi- CM S zosaccharomyces pombe.
21. bgll-genens nukleotidsekvens, vilken hela sek-vens eller en del därav är stämplad att användas som probe, kännetecknad av att bgll-genen har en nukleotidsekvens enligt figur 1.
22. Användning av en probe omfattande nukleotid- sekvensen enligt patentkrav 21 för identifiering av en β-glukosidasgen som härstammar frän filamentmögel.
23. Användning enligt patentkrav 22, känne tecknad av att proben i fraga har erhällits frän Tri- 10 choderma reesei.
24. Användning enligt patentkrav 22 eller 23, kännetecknad av att en eller flera prober kan användas för identifiering av nämnda β-glukosidasgen frän fila- mentmögel. δ CM CO O 00 CM X cc a. 00 00 CD CM 00 O)
FI932633A 1990-12-10 1993-06-09 Förbättrad sackarifikation av cellulosa genom kloning och amplifikering av b-glukosidasgenen av Trichoderma reesei FI121925B (sv)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US62514090A 1990-12-10 1990-12-10
US62514090 1990-12-10
PCT/US1991/009285 WO1992010581A1 (en) 1990-12-10 1991-12-10 IMPROVED SACCHARIFICATION OF CELLULOSE BY CLONING AND AMPLIFICATION OF THE β-GLUCOSIDASE GENE OF TRICHODERMA REESEI
US9109285 1991-12-10

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FI932633A0 FI932633A0 (fi) 1993-06-09
FI932633L FI932633L (fi) 1993-06-09
FI121925B true FI121925B (sv) 2011-06-15

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US (1) US6022725A (sv)
EP (3) EP2075338A3 (sv)
JP (2) JPH06503960A (sv)
AT (2) ATE423207T1 (sv)
CA (1) CA2097180C (sv)
DE (2) DE69133100T3 (sv)
DK (2) DK1225227T3 (sv)
ES (2) ES2322032T3 (sv)
FI (1) FI121925B (sv)
WO (1) WO1992010581A1 (sv)

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EP1225227A1 (en) 2002-07-24
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DE69133100D1 (de) 2002-10-10
DE69133100T3 (de) 2015-09-03
JPH06503960A (ja) 1994-05-12
WO1992010581A1 (en) 1992-06-25
EP2075338A2 (en) 2009-07-01
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EP1225227B1 (en) 2009-02-18
CA2097180C (en) 2007-08-14
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ATE223490T1 (de) 2002-09-15
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US6022725A (en) 2000-02-08
EP0562003B2 (en) 2015-04-01
DE69133100T2 (de) 2003-05-22
ES2182818T3 (es) 2003-03-16
DK1225227T3 (da) 2009-06-22
CA2097180A1 (en) 1992-06-11
ATE423207T1 (de) 2009-03-15
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