DK2935601T3 - Rekombinante mikrobeceller, der fremstiller mindst 28 % eicosapentaensyre som tørvægt - Google Patents

Rekombinante mikrobeceller, der fremstiller mindst 28 % eicosapentaensyre som tørvægt Download PDF

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DK2935601T3
DK2935601T3 DK13821271.7T DK13821271T DK2935601T3 DK 2935601 T3 DK2935601 T3 DK 2935601T3 DK 13821271 T DK13821271 T DK 13821271T DK 2935601 T3 DK2935601 T3 DK 2935601T3
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ala
epa
gly
val
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DK13821271.7T
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Quinn Qun Zhu
Seung-Pyo Hong
Pamela L Sharpe
Narendra S Yadav
Michael Dauner
Xiaochun Fan
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Du Pont
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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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • C12P7/6432—Eicosapentaenoic acids [EPA]
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • 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, e.g. protozoa; 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
    • C12N1/16—Yeasts; Culture media therefor
    • 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.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
    • C12P7/6431—Linoleic acids [18:2[n-6]]
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6463—Glycerides obtained from glyceride producing microorganisms, e.g. single cell oil
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6472—Glycerides containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
    • 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)
    • C12Y203/01023—1-Acylglycerophosphocholine O-acyltransferase (2.3.1.23), i.e. lysophosphatidylcholine acyltransferase or LPCAT
    • 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
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts

Claims (10)

  1. REKOMBINANTE MIKROBECELLER, DER FREMSTILLER MINDST 28 % EICOS APENTAENS YRE SOM T0RV7EGT E Rekombinant Yarrowia-celle, der fremstiller en olie, der indeholder mindst 28 % eicosapentaensyre (EPA) malt som en vsgtprocent af tprcellevsgt, hvor cellen omfatter: a) mindst én polynukleotidsekvens, der koder for en acyl-CoA:lysophosphatidylcholin-acyltransferase (LPCAT) med en aminosyresekvens, der er mindst 90 % identisk med sekvensen ifplge SEQ ID NO: 40 (YILPCAT), og b) LPCAT omfatter mutationer ved (i) aminosyreposition 136 ved asndring af methionin til en anden aminosyre, og (ii) aminosyreposition 389 ved asndring af threonin til en anden aminosyre, hvor LPCAT har LPCAT-aktivitet, og hvor polynukleotidet, der koder for LPCAT, er operativt forbundet med mindst én regulatorisk sekvens, og hvor cellen endvidere omfatter en nedregulering af en endogen polynukleotidsekvens, der koder for Sou2 sorbitol-anvendelsesprotein.
  2. 2. Rekombinant Yarrowia-celle ifplge krav 1, hvor LPCAT er en Yarrowia lipolytica LPCAT med en sekvens ifplge SEQ ID NO: 40.
  3. 3. Rekombinant Yarrowia-ce\\e ifplge krav 1, hvor nedreguleringen skyldes en mutation af den endogene polynukleotidsekvens, og hvor mutationer er udvalgt fra grappen bestaende af en substitution, deletion og insertion.
  4. 4. Rekombinant Yarrowia-ceïle ifplge krav 3, hvor Sou2 sorbitol-anvendelsesprotein omfatter en aminosyresekvens, der er mindst 95 % identisk med SEQ ID NO: 10.
  5. 5. Rekombinant Yarrowia-ce\\e ifplge krav 3 eller 4, hvor nedreguleringen reducerer den totale masngde sukkeralkoholer fremstillet af den mikrobielle celle.
  6. 6. Rekombinant Yarrowia-ce\\e ifplge et hvilket som helst af kravene 1 til 5, hvor cellen endvidere omfatter: (a) mindst én polynukleotidsekvens, der koder for phospholipid: diacylglycerol-acyltransferase (PDAT), (b) mindst én polynukleotidsekvens, der koder for delta-12-desaturase, og (c) mindst én polynukleotidsekvens, der koder for et dihomo-gamma-linolensyre-(DGLA) syntasemultizym; hvor hver af polynukleotidsekvenserne fra (a)-(c) er operativt forbundet med mindst én regulatorisk sekvens.
  7. 7. Rekombinant Yarrowia-ce\\e ifplge krav 6, hvor DGLA-syntasemultizymet omfatter en delta-9 elongase bundet til en delta-8-desaturase.
  8. 8. Rekombinant Yarrowia-ce\\e ifplge et hvilket som helst af kravene 1 til 7, hvor cellen endvidere omfatter: (a) mindst én polynukleotidsekvens, der koder for delta-8-desaturase, (b) mindst én polynukleotidsekvens, der koder for malonyl-CoA-syntetase (MCS), og (c) mindst én polynukleotidsekvens, der koder for acyl-CoA:lysophosphatidsyre-acyltransferase (LPAAT), hvor hver af polynukleotidsekvenserne fra (a)-(c) er operativt forbundet med mindst én regulatorisk sekvens.
  9. 9. Rekombinant Yarrowia-ce\\e ifplge et hvilket som helst af kravene 1 til 8, hvor oliën omfatter mindst 30 % EPA malt som et vsgtforhold af vasrtscellens tprvsgt.
  10. 10. Fremgangsmade til fremstilling af en mikrobiel olie, der omfatter eicosapentaensyre, hvilken fremgangsmade omfatter: a) dyrkning af den rekombinante Yarrowia-celle ifplge et hvilket som helst af kravene 1 til 9, hvor der fremstilles en mikrobiel olie, der omfatter eicosapentaensyre, og b) eventuel gen vinding af den mikrobielle olie fra trin (a).
DK13821271.7T 2012-12-21 2013-12-18 Rekombinante mikrobeceller, der fremstiller mindst 28 % eicosapentaensyre som tørvægt DK2935601T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201261740502P 2012-12-21 2012-12-21
PCT/US2013/075896 WO2014100062A1 (en) 2012-12-21 2013-12-18 Recombinant microbial cells that produce at least 28% eicosapentaenoic acid as dry cell weight

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US (1) US9701989B2 (da)
EP (1) EP2935601B1 (da)
JP (1) JP2016502852A (da)
BR (1) BR112015014781A2 (da)
CA (1) CA2893409A1 (da)
CL (1) CL2015001673A1 (da)
DK (1) DK2935601T3 (da)
NO (1) NO2976143T3 (da)
WO (1) WO2014100062A1 (da)

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AU2015343307B2 (en) 2014-11-06 2021-05-20 Iff Us Holding, Llc Peptide-mediated delivery of RNA-guided endonuclease into cells
DK3362560T3 (da) 2015-10-12 2022-11-07 Dupont Us Holding Llc Beskyttede dna-templater til genmodificering og forøget homolog rekombination i celler og fremgangsmåder til anvendelse
CN108330114B (zh) * 2018-03-21 2020-11-03 北京大学 一种利用epa的甘油二酯酰基转移酶及其应用
KR102536297B1 (ko) * 2021-02-08 2023-05-24 세종대학교산학협력단 식물체의 오메가 3 지방산 함량 증가 방법

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Publication number Publication date
NO2976143T3 (da) 2018-08-04
EP2935601A1 (en) 2015-10-28
JP2016502852A (ja) 2016-02-01
WO2014100062A1 (en) 2014-06-26
CA2893409A1 (en) 2014-06-26
BR112015014781A2 (pt) 2017-10-10
CL2015001673A1 (es) 2015-10-23
US20140186906A1 (en) 2014-07-03
US9701989B2 (en) 2017-07-11
EP2935601B1 (en) 2018-03-21

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