DK2140006T3 - Delta-5-desaturaser og anvendelse heraf til fremstilling af polyumættede fedtsyrer - Google Patents

Delta-5-desaturaser og anvendelse heraf til fremstilling af polyumættede fedtsyrer Download PDF

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Publication number
DK2140006T3
DK2140006T3 DK08747327.8T DK08747327T DK2140006T3 DK 2140006 T3 DK2140006 T3 DK 2140006T3 DK 08747327 T DK08747327 T DK 08747327T DK 2140006 T3 DK2140006 T3 DK 2140006T3
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aat
desaturase
leu
ggt
seq
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DK08747327.8T
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English (en)
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Howard Glenn Damude
Quinn 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
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/02—Other edible oils or fats, e.g. shortenings, cooking oils characterised by the production or working-up
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • 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
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • C12N9/0083—Miscellaneous (1.14.99)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/19—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with oxidation of a pair of donors resulting in the reduction of molecular oxygen to two molecules of water (1.14.19)

Claims (13)

  1. DELTA-5-DES ATURASER OG ANVENDELSE HERAF TIL FREMSTILLING AF POLYUMÆTTEDE FEDTSYRER
    1. Mikrobiel værtscelle omfattende et vektorkonstrukt, der omfatter: (a) en nukleotidsekvens, der koder for et polypeptid, der har A5-desaturaseaktivitet, hvor polypeptidet er mindst 80 % amino syreidentisk, baseret på Clustal V-alignmentmetoden, sammenlignet med amino syre sekvensen som vist i SEQ ID NO: 13; eller (b) en nukleotidsekvens, der koder for et polypeptid, der har A5-desaturaseaktivitet, hvor nukleotidsekvensen er mindst 80 % sekvensidentisk, baseret på BLASTN-alignmentmetoden, sammenlignet med nukleotidsekvensen som vist i SEQ ID NO: 12 eller SEQ ID NO: 25.
  2. 2. Mikrobiel værtscelle ifølge krav 1, hvor polypeptidet, der har A5-desaturaseaktivitet, omfatter amino syre sekvensen som vist i SEQ ID NO: 13.
  3. 3. Mikrobiel værtscelle ifølge krav 1, hvor den mikrobielle værtscelle er udvalgt fra gruppen bestående af gær, alger, bakterier, euglenider, stramenofiler og svampe.
  4. 4. Mikrobiel værtscelle ifølge krav 3, hvor cellen er en svamp af arten Mortierella.
  5. 5. Mikrobiel værtscelle ifølge krav 3, hvor cellen er en stramenofil, der er udvalgt fra gruppen bestående af: Thraustochytrium og Schizochytrium.
  6. 6. Mikrobiel værtscelle ifølge krav 3, hvor cellen er en olieholdig gær.
  7. 7. Mikrobiel værtscelle ifølge krav 6, hvor den olieholdige gær er udvalgt fra gruppen bestående af: Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon og Lipomyces.
  8. 8. Mikrobiel værtscelle ifølge krav 7, hvor den olieholdige gær er Yarrowia.
  9. 9. Fremgangsmåde til fremstilling af arachidonsyre, hvilken fremgangsmåde omfatter: a) tilvejebringelse af en mikrobiel værtscelle, der omfatter: (i) et vektorkonstrukt, der omfatter et nukleotidmolekyle, der koder for et Δ5- desaturasepolypeptid, der er mindst 80 % aminosyreidentisk sammenlignet med et polypeptid, der har amino syre sekvensen som vist i SEQ ID NO: 13, baseret på Clustal V-alignmentmetoden; og (ii) en dihomo-y-linolensyrekilde; b) dyrkning af den mikrobielle værtscelle fra trin (a) under forhold, hvor nukleotidmolekylet, der koder for A5-desaturasepolypeptidet, udtrykkes, og dihomo-y-linolensyren omdannes til arachidonsyre; og c) eventuelt indvinding af arachidonsyren fra trin (b).
  10. 10. Fremgangsmåde til fremstilling af eicosapentaensyre, hvilken fremgangsmåde omfatter: a) tilvejebringelse af en mikrobiel værtscelle, der omfatter: (i) et vektorkonstrukt, der omfatter et nukleotidmolekyle, der koder for et Δ5- desaturasepolypeptid, der er mindst 80 % aminosyreidentisk sammenlignet med et polypeptid, der har amino syre sekvensen som vist i SEQ ID NO: 13, baseret på Clustal V-alignmentmetoden; og (ii) en eicosatetraensyrekilde; b) dyrkning af den mikrobielle værtscelle fra trin (a) under forhold, hvor nukleotidmolekylet, der koder for A5-desaturasepolypeptidet, udtrykkes, og eicosatetraensyren omdannes til eicosapentaensyre; og c) eventuelt indvinding af eicosapentaensyren fra trin (b).
  11. 11. Fremgangsmåde ifølge et hvilket som helst af kravene 9 eller 10, hvor den mikrobielle værtscelle er en Yarrowia, og hvor A5-desaturasepolypeptidet omfatter SEQ ID NO: 13.
  12. 12. Fremgangsmåde ifølge et hvilket som helst af kravene 9 eller 10, hvor: a) nukleotidmolekylet har nukleinsyresekvensen udvalgt fra gruppen bestående af SEQ ID NO: 12 og SEQ ID NO: 25; og b) værtscellen er Yarrowia lipolytica.
  13. 13. Nukleinsyremolekyle, der koder for A5-desaturase som vist i SEQ ID NO: 26, hvor mindst 174 codoner af nukleinsyremolekylet er codon-optimeret til ekspression i Yarrowia.
DK08747327.8T 2007-05-03 2008-05-01 Delta-5-desaturaser og anvendelse heraf til fremstilling af polyumættede fedtsyrer DK2140006T3 (da)

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PCT/US2008/062196 WO2008137532A1 (en) 2007-05-03 2008-05-01 Delta-5 desaturases and their use in making polyunsaturated fatty acids

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US (4) US7943365B2 (da)
EP (3) EP2140007B1 (da)
AU (1) AU2008247766B2 (da)
BR (1) BRPI0809877A2 (da)
CA (1) CA2684719C (da)
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WO (2) WO2008137516A1 (da)

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