DK2235190T3 - Sammensætninger og fremgangsmåder til fremstilling af isopren - Google Patents

Sammensætninger og fremgangsmåder til fremstilling af isopren Download PDF

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DK2235190T3
DK2235190T3 DK08860589.4T DK08860589T DK2235190T3 DK 2235190 T3 DK2235190 T3 DK 2235190T3 DK 08860589 T DK08860589 T DK 08860589T DK 2235190 T3 DK2235190 T3 DK 2235190T3
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Prior art keywords
isoprene
cells
polypeptide
nucleic acid
dxs
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DK08860589.4T
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English (en)
Inventor
Marguerite Cervin
Gregory M Whited
Gopal K Chotani
Fernando Valle
Carol Fioresi
Karl J Sanford
Joseph C Mcauliffe
Frank J Feher
Aaron S Puhala
Andrei Miasnikov
Ilana S Aldor
Duca Richard La
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Danisco Us Inc
Goodyear Tire & Rubber
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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
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/007—Preparation of hydrocarbons or halogenated hydrocarbons containing one or more isoprene units, i.e. terpenes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1022—Transferases (2.) transferring aldehyde or ketonic groups (2.2)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1205—Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1229—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • 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
    • C12N9/88—Lyases (4.)
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    • 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
    • C12N9/90—Isomerases (5.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y203/00—Acyltransferases (2.3)
    • C12Y203/01—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • C12Y203/01009—Acetyl-CoA C-acetyltransferase (2.3.1.9)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y207/00—Transferases transferring phosphorus-containing groups (2.7)
    • C12Y207/01—Phosphotransferases with an alcohol group as acceptor (2.7.1)
    • C12Y207/01036—Mevalonate kinase (2.7.1.36)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y401/00—Carbon-carbon lyases (4.1)
    • C12Y401/01—Carboxy-lyases (4.1.1)
    • C12Y401/01033—Diphosphomevalonate decarboxylase (4.1.1.33), i.e. mevalonate-pyrophosphate decarboxylase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y402/00—Carbon-oxygen lyases (4.2)
    • C12Y402/03—Carbon-oxygen lyases (4.2) acting on phosphates (4.2.3)
    • C12Y402/03027—Isoprene synthase (4.2.3.27)
    • C—CHEMISTRY; METALLURGY
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    • C12Y—ENZYMES
    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/03—Intramolecular oxidoreductases (5.3) transposing C=C bonds (5.3.3)
    • C12Y503/03002—Isopentenyl-diphosphate DELTA-isomerase (5.3.3.2)
    • 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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  • Biomedical Technology (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (13)

1. Bakterie-, svampe-, gær- eller planteceller omfattende en eller flere nukleinsyrer, som koder for: (a) et isopren-syntase-polypeptid, hvor isopren-syntase-polypeptidet er kodet afen heterolog nukleinsyre, og (b) et isopentenyl-diphosphat-delta-isomerase-(IDI)-polypeptid, hvor IDI-polypeptidet er kodet af en heterolog nukleinsyre; og/eller hvor IDI-polypeptidet er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et IDI-polypeptid og (c) (i) et l-Deoxyxylulose-5-phosphat-syntase-(DXS)-polypeptid, hvor DXS-polypeptidet er kodet afen heterolog nukleinsyre; og/eller hvor DXS-polypeptidet er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et DXS-polypeptid og/eller (ii) et eller flere mevalonat-(MVA)-signalvej-polypeptider, hvor nævnte ene eller flere MVA-signalvej-polypeptider er kodet af en heterolog nukleinsyre; og/eller hvor nævnte ene eller flere MVA-signalvej-polypeptider er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et eller flere MVA-signalvej-polypeptider, hvor cellerne producerer mere end omkring 400 nrnol/gwcm/tirne af isopren.
2. Bakterie-, svampe-, gær- eller planteceller omfattende en eller flere nukleinsyrer, som koder for: (a) et isopren-syntase-polypeptid, hvor isopren-syntase-polypeptidet er kodet afen heterolog nukleinsyre, og (b) et isopentenyl-diphosphat-delta-isomerase-(IDI)-polypeptid, hvor IDI-polypeptidet er kodet af en heterolog nukleinsyre; og/eller hvor IDI-polypeptidet er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et IDI-polypeptid og (C) (i) et l-Deoxyxylulose-S-phosphat-syntase-(DXS)-polypeptid hvor DXS-polypeptidet er kodet af en heterolog nukleinsyre; og/eller hvor DXS-polypeptidet er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et DXS-polypeptid og/eller (ii) et eller flere mevalonat-(MVA)-signalvej-polypeptider, hvor nævnte ene eller flere MVA-signalvej-polypeptider er kodet af en heterolog nukleinsyre; og/eller hvor nævnte ene eller flere MVA-signalvej-polypeptider er kodet af en indsættelse af en kopi af en endogen nukleinsyre, som koder for et eller flere MVA-signalvej-polypeptider, hvor cellerne omdanner mere end omkring 0,2 molprocent af kulstoffet, som cellerne forbruger fra et celledyrkningsmedium, til isopren.
3. Cellerne ifølge krav 1 eller krav 2, hvor den heterologe nukleinsyre er funktionelt forbundet til en promoter.
4. Cellerne ifølge et hvilket som helst af kravene 1-3, hvor isopren-syntase-polypeptidet er et plante-isopren-syntase-polypeptid.
5. Cellerne ifølge et hvilket som helst af kravene 1-4, hvor MVA-signalvej-polypeptidet er en mevalonat-kinase (MVK), hvor MVK eventuelt er et polypeptid fra Methanosarcina.
6. Cellerne ifølge et hvilket som helst af kravene 1-5, hvor isopren-syntase-polypeptidet er et naturligt forekommende polypeptid fra Pueraria eller Populus.
7. Cellerne ifølge et hvilket som helst af kravene 1-6, hvor cellerne er bakterieceller, eventuelt Bacillus-, Escherichia- eller Pantoea-celler, eller hvor cellerne er svampeceller, eventuelt Trichoderma-ceUer, eller hvor cellerne er gærceller, eventuelt Yarrowia- eller Saccharomyces-ceUer.
8. Fremgangsmåde til fremstilling af isopren, hvilken fremgangsmåde omfatter at dyrke celler ifølge et hvilket som helst af kravene 1-7 under betingelser tilstrækkelige til fremstilling af isopren.
9. Fremgangsmåden ifølge krav 8, hvilken fremgangsmåde omfatter: (a) at dyrke celler omfattende en heterolog nukleinsyre som koder for et isopren-syntase-polypeptid under egnede dyrkningsbetingelser til fremstillingen af isopren, hvor cellerne producerer mere end omkring 400 nmol/g wcm /time af isopren, og (b) at fremstille isopren.
10. Fremgangsmåden ifølge krav 8, hvilken fremgangsmåde omfatter: (a) at dyrke celler omfattende en heterolog nukleinsyre som koder for et isopren-syntase-polypeptid under egnede dyrkningsbetingelser til fremstillingen af isopren, hvor cellerne omdanner mere end omkring 0,2 molprocent af kulstoffet, som cellerne forbruger fra et celledyrkningsmedium, til isopren, og (b) at fremstille isopren.
11. Fremgangsmåden ifølge et hvilket som helst af kravene 8-10, yderligere omfattende at genvinde isoprenen fremstillet af cellerne.
12. Anvendelse af celler ifølge et hvilket som helst af kravene 1-7 i fremstillingen af isopren.
13. Fremgangsmåden ifølge et hvilket som helst af kravene 8-10, yderligere omfattende trinnet at oprense isoprenen fremstillet af cellerne, og/eller omfattende trinnet at polymerisere isoprenen fremstillet af cellerne.
DK08860589.4T 2007-12-13 2008-12-15 Sammensætninger og fremgangsmåder til fremstilling af isopren DK2235190T3 (da)

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US1357407P 2007-12-13 2007-12-13
PCT/US2008/086869 WO2009076676A2 (en) 2007-12-13 2008-12-15 Compositions and methods for producing isoprene

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US (5) US8288148B2 (da)
EP (2) EP3354740A1 (da)
JP (3) JP6049982B2 (da)
KR (1) KR20100118973A (da)
CN (2) CN102027124B (da)
AU (1) AU2008334945C1 (da)
BR (1) BRPI0823506A8 (da)
CA (1) CA2709107A1 (da)
DK (1) DK2235190T3 (da)
MX (1) MX2010006576A (da)
RU (1) RU2545699C2 (da)
SG (2) SG191671A1 (da)
WO (1) WO2009076676A2 (da)

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