DK2678432T3 - Rekombinante mikroorganismer og anvendelser deraf - Google Patents

Rekombinante mikroorganismer og anvendelser deraf Download PDF

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DK2678432T3
DK2678432T3 DK12749472.2T DK12749472T DK2678432T3 DK 2678432 T3 DK2678432 T3 DK 2678432T3 DK 12749472 T DK12749472 T DK 12749472T DK 2678432 T3 DK2678432 T3 DK 2678432T3
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Sean Dennis Simpson
Michael Koepke
Fungmin Liew
Wendy Yiting Chen
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Lanzatech New Zealand Ltd
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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
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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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
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    • 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
    • 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
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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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26—Ketones
    • C12P7/28—Acetone-containing products
    • C12P7/30—Acetone-containing products produced from substrate containing inorganic compounds other than water
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01001—Alcohol dehydrogenase (1.1.1.1)
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
    • C12Y101/01002—Alcohol dehydrogenase (NADP+) (1.1.1.2), i.e. aldehyde reductase
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    • 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)
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    • C12Y401/00—Carbon-carbon lyases (4.1)
    • C12Y401/01—Carboxy-lyases (4.1.1)
    • C12Y401/01004—Acetoacetate decarboxylase (4.1.1.4)
    • 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)
    • C12Y401/01074—Indolepyruvate decarboxylase (4.1.1.74)
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/10—Process efficiency
    • Y02P20/133—Renewable energy sources, e.g. sunlight

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
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Claims (11)

1. Fremgangsmåde til fremstilling af isopropanol ved mikrobiel fermentering, der omfatter fermentering af et substrat, der omfatter CO, ved anvendelse af en rekombinant carboxydotrof eddikesyredannende mikroorganisme, der er i stand til at frembringe isopropanol ved fermentation af CO, hvor mikroorganismen udtrykker enzymerne: acetyl-coenzym A-acetyltransferase (thiolase; ThlA; E.C. 2.3.1.9); acetoacetyl-CoA:acetat-coenzym A-transferase A (CoA-transferase; CtfA; EC 2.8.3.9); og acetoacetyl-CoA:acetat-coenzym A-transferase B (CoA-transferase; CtfB; EC 2.8.3.9); og hvor mikroorganismen endvidere udtrykker enzymet: a. acetoacetat-decarboxylase (Adc; EC 4.1.1.4) eller b. alpha-ketoisovalerat-decarboxylase (decarboxylase; KivD; EC4.1.1.74); hvor enzymerne ikke er naturligt til stede i en parental mikroorganisme, hvoraf den rekombinante mikroorganisme er afledt.
2. Fremgangsmåde ifølge krav la, hvor mikroorganismen endvidere udtrykker enzymet alkoholdehydrogenase (Adh; EC 1.1.1.2).
3. Fremgangsmåde ifølge krav 2, hvor Adh udtrykkes eksogent.
4. Fremgangsmåde ifølge krav la, hvor mikroorganismen endvidere udtrykker enzymet alkoholdehydrogenase (Adh2; EC 1.1.1.1).
5. Fremgangsmåde ifølge krav 4, hvor Adh2 udtrykkes eksogent.
6. Fremgangsmåde ifølge krav la, hvor mikroorganismen endvidere udtrykker enzymerne alpha-ketoisovalerat-decarboxylase (decarboxylase; KivD; EC4.1.1.74) og alkoholdehydrogenase (Adh2; EC 1.1.1.1).
7. Fremgangsmåde ifølge krav 6, hvor Adh2 udtrykkes eksogent.
8. Fremgangsmåde ifølge krav 1, hvor den parentale mikroorganisme vælges fra gruppen af carboxydotrofe eddikesyredannende bakterier, der omfatter: Clostridium autoethanogenum, Clostridium ljungdahlii, Clostridium ragsdalei, Clostridium carboxidivorans, Clostridium drakei, Clostridium scatologenes, Butyribacterium limosum, Butyribacterium methylotrophicum, Acetobacterium woodii, Alkalibaculum bacchii, Blautia producta, Eubacterium limosum, Moorella thermoacetica, Moorella thermautotrophica, Oxobacter pfetmigii og Thermoanaerobacter kiuvi.
9. Fremgangsmåde ifølge krav 6, hvor parentale mikroorganisme er Clostridium autoethanogenum DSM23693 eller Clostridium ljungdahlii DSM13528 (eller ATCC55383).
10. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor enzymet: a. ThlA er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 18; og b. CtfA er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 19; og c. CtfB er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 20 og/eller d. Adc er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 21; og/eller e. KivD er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 71; og/eller f. Adh er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 2, 3 eller 4; og/eller g. Adh2 er kodet for af et nukleotid, der omfatter sekvensen ifølge SEQ ID NO: 75.
11. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor: a. det gasformige substrat, der omfatter CO, fremstilles ved en industriel proces valgt fra gruppen bestående af fremstilling af jernholdige metalprodukter, fremstilling af ikke-jemholdige produkter, olieraffineringsprocesser, forgasning af kul, forgasning af biomasse, elkraftproduktion, carbon black-produktion og koksfremstilling; eller b. det gasformige substrat omfatter 20 % til 70 % CO efter volumen.
DK12749472.2T 2011-02-25 2012-02-24 Rekombinante mikroorganismer og anvendelser deraf DK2678432T3 (da)

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JP (1) JP6199747B2 (da)
KR (2) KR102015672B1 (da)
CN (2) CN108486169B (da)
AU (1) AU2012221176B2 (da)
BR (1) BR112013021524A2 (da)
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KR102121886B1 (ko) 2020-06-15
EP2678432B1 (en) 2018-08-01
AU2012221176A1 (en) 2013-05-09
KR20190097292A (ko) 2019-08-20
KR102015672B1 (ko) 2019-08-29
CN108486169B (zh) 2022-08-12
TW201309795A (zh) 2013-03-01
PL2678432T4 (pl) 2019-05-31
JP2014506478A (ja) 2014-03-17
AU2012221176B2 (en) 2015-03-19
EP3401405A1 (en) 2018-11-14
BR112013021524A2 (pt) 2017-06-13
CA2825267A1 (en) 2012-08-30
MY193665A (en) 2022-10-24
CN103502435B (zh) 2018-05-18
TWI541348B (zh) 2016-07-11
WO2012115527A2 (en) 2012-08-30
CA2995872A1 (en) 2012-08-30
NZ614459A (en) 2015-09-25
KR20140012106A (ko) 2014-01-29
EP2678432A2 (en) 2014-01-01
CN108486169A (zh) 2018-09-04
JP6199747B2 (ja) 2017-09-20
CA2825267C (en) 2018-04-03
EP2678432A4 (en) 2016-06-22
CA2995872C (en) 2019-09-17
PL2678432T3 (pl) 2019-05-31
CN103502435A (zh) 2014-01-08
ES2702621T3 (es) 2019-03-04
MY168208A (en) 2018-10-15

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