DK2443248T3 - FORBEDRING AF LANGKÆDET POLYUMÆTTET OMEGA-3- OG OMEGA-6- FEDTSYREBIOSYNTESE VED EKSPRESSION AF ACYL-CoA-LYSOPHOSPHOLIPID- ACYLTRANSFERASER - Google Patents

FORBEDRING AF LANGKÆDET POLYUMÆTTET OMEGA-3- OG OMEGA-6- FEDTSYREBIOSYNTESE VED EKSPRESSION AF ACYL-CoA-LYSOPHOSPHOLIPID- ACYLTRANSFERASER Download PDF

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DK2443248T3
DK2443248T3 DK10790004.5T DK10790004T DK2443248T3 DK 2443248 T3 DK2443248 T3 DK 2443248T3 DK 10790004 T DK10790004 T DK 10790004T DK 2443248 T3 DK2443248 T3 DK 2443248T3
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lys
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phe
ser
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DK10790004.5T
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Narendra S Yadav
Hongxiang Zhang
Qun Zhu
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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
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6481—Phosphoglycerides
    • 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
    • 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
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
    • 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
    • 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
    • C12P7/6409—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
    • C12P7/6434—Docosahexenoic acids [DHA]
    • 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

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
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  • Biomedical Technology (AREA)
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  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fats And Perfumes (AREA)

Claims (6)

  1. DK/EP 2443248 T3 -1- FORBEDRING AF LANGKÆDET POLYUMÆTTET OMEGA-3- OG OMEGA-6-FEDTSYREBIOSYNTESE VED EKSPRESSION AF ACYL-CoA-LYSOPHOSPHOLIPID- AC YLTR AN SFER ASER 5 E Rekombinant olieholdig eicosapentaensyre- (EPA) producerende Yarrowia //po/yt/ca-værtscelle, der omfatter mindst ét transgen, der omfatter et polynukleotid, der koder for et polypeptid med mindst acyl-CoAdysophospholipid-acyltransferaseaktivitet, hvor polypeptidet har mindst 95 % aminosyreidentitet, baseret pä Clustal W-alignmentmetoden ved sammenligning med SEQ ID NO: 11; hvor: 10 transgenet omfatter polynukleotidet, der er operativt bundet til mindst én regulatorisk sekvens, hvor den mindst ene regulatoriske sekvens indbefatter en promoter, der er en konstitutiv promoter, eller der er en regulerbar promoter med en inducerbar aktivitet, der muliggpr induceret ekspression; og hvor polypeptidet er overudtrykt ved sammenligning med en kontrolværtscelle.
  2. 2. Rekombinant værtscelle ifplge krav 1, hvor polynukleotidet, der koder for et polypeptid med 15 mindst acyl-CoA:lysophospholipid-acyltransferaseaktivitet, er stabilt integreret i vaertscellen.
  3. 3. Rekombinant værtscelle ifplge krav 1 eller 2, hvor promoteren er en konstitutiv promoter.
  4. 4. Rekombinant værtscelle ifplge krav 1 eller 2, hvor promoteren er en regulerbar promoter med inducerbar aktivitet.
  5. 5. Fremgangsmâde til fremstilling af en olie, der omfatter eicosapentaensyre, hvilken fremgangsmâde 20 omfatter: a) dyrkning af den olieholdige Yarrowia lipolytica-værtscelle ifplge krav 2, hvor værtscellen har en stigning i Ci8. til C2o-elongeringsomdannelseseffektivitet pâ mindst 13 % sammenlignet med en kontrolværtscelle, og hvor der fremstilles en olie omfattende eicosapentaensyre; og b) eventuel genvinding af den mikrobielle olie fra trin (a).
  6. 6. Fremgangsmâde ifplge krav 5, hvor den genvundne olie fra trin (b) forarbejdes yderligere.
DK10790004.5T 2009-06-16 2010-06-14 FORBEDRING AF LANGKÆDET POLYUMÆTTET OMEGA-3- OG OMEGA-6- FEDTSYREBIOSYNTESE VED EKSPRESSION AF ACYL-CoA-LYSOPHOSPHOLIPID- ACYLTRANSFERASER DK2443248T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US18735909P 2009-06-16 2009-06-16
PCT/US2010/038527 WO2010147900A1 (en) 2009-06-16 2010-06-14 IMPROVEMENT OF LONG CHAIN OMEGA-3 AND OMEGA-6 POLYUNSATURATED FATTY ACID BIOSYNTHESIS BY EXPRESSION OF ACYL-CoA LYSOPHOSPHOLIPID ACYLTRANSFERASES

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DK2443248T3 true DK2443248T3 (da) 2018-03-12

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Country Status (10)

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EP (1) EP2443248B1 (da)
JP (1) JP5931721B2 (da)
CN (1) CN102459627B (da)
AU (1) AU2010260227B2 (da)
BR (1) BRPI1009690A2 (da)
CA (1) CA2763424C (da)
CL (1) CL2011003161A1 (da)
DK (1) DK2443248T3 (da)
NO (1) NO2443248T3 (da)
WO (1) WO2010147900A1 (da)

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Publication number Publication date
EP2443248A1 (en) 2012-04-25
JP2012529912A (ja) 2012-11-29
AU2010260227A1 (en) 2011-11-10
CL2011003161A1 (es) 2012-05-25
JP5931721B2 (ja) 2016-06-08
EP2443248A4 (en) 2013-07-10
WO2010147900A1 (en) 2010-12-23
CA2763424C (en) 2018-06-12
AU2010260227B2 (en) 2015-01-29
EP2443248B1 (en) 2017-12-27
CA2763424A1 (en) 2010-12-23
CN102459627B (zh) 2015-07-29
BRPI1009690A2 (pt) 2015-08-25
CN102459627A (zh) 2012-05-16
NO2443248T3 (da) 2018-05-26

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