DK2970926T3 - Genmanipulering af mikroorganismer - Google Patents

Genmanipulering af mikroorganismer Download PDF

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Publication number
DK2970926T3
DK2970926T3 DK14713925.7T DK14713925T DK2970926T3 DK 2970926 T3 DK2970926 T3 DK 2970926T3 DK 14713925 T DK14713925 T DK 14713925T DK 2970926 T3 DK2970926 T3 DK 2970926T3
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optionally
gene
nucleic acid
desaturase
seq
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DK14713925.7T
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Mark A Scaife
Roberto E Armenta
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Dsm Nutritional Products Ag
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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
    • 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
    • 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)
    • 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
    • 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)
    • C12Y114/19006—DELTA12-fatty-acid desaturase (1.14.19.6), i.e. oleoyl-CoA DELTA12 desaturase

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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)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Claims (6)

10. Genmanipuleret mikroorganisme ifølge krav 6, hvor det mindst ene polyumættede fedtsyre-biosyntese-polynukleotid er endogent for mikroorganismen, eller hvor det mindst ene polyumættede fedtsyre-biosyntese-polynukleotid er eksogent for mikroorganismen, hvor det mindst ene polyumættede fedtsyre-biosyntese-polynukleotid valgfrit koder for et enzympolypeptid eller en del af et enzympolypeptidkompleks, som er valgt fra gruppen bestående af fedtsyresynthase (FAS), A5-elongase, A5-desaturase, A12-desaturase, A4-desaturase og polyketid-PUFA-synthase (PKS).
11. Fremgangsmåde til fremstilling af polyumættede fedtsyrer, hvilken fremgangsmåde omfatter: tilvejebringelse af en genmanipuleret Thraustochytrium-ceWe, hvor den genmanipulerede Thraustochytrium-ceWe udtrykker et heterologt nukleinsyremolekyle med en sekvens, der er mindst 90 % identisk med nukleinsyresekvensen ifølge SEQ ID NO:70, som koder for et polypeptid med Δ12-desaturase-aktivitet; hvor den heterologe ekspression ændrer produktionen af én eller flere polyumættede fedtsyrer via den genmanipulerede celle sammenlignet med en referencecelle, når den genmanipulerede celle og referencecellen dyrkes under sammenlignelige betingelser; og dyrkning af den genmanipulerede Thraustochytrium-ceWe under betingelser og i et tidsrum, der er tilstrækkeligt til at opnå produktion af den eller de polyumættede fedtsyrer.
12. Fremgangsmåde ifølge krav 11, der endvidere omfatter ekspression af mindst ét heterologt polynukleotid, som koder for et enzympolypeptid eller en del af et enzympolypeptidkompleks, der er valgt fra gruppen bestående af fedtsyresynthase (FAS), A5-elongase, A5-desaturase, A4-desaturase og polyketid-PUFA-synthase (PKS).
13. Fremgangsmåde til fremstilling af polyumættede fedtsyrer, hvilken fremgangsmåde omfatter: tilvejebringelse af en genmanipuleret Thraustochytrium-ceWe, hvor den genmanipulerede Thraustochytrium-ceWe udtrykker mindst ét polyumættet fedtsyre-biosyntese-polynukleotid, der er operativt forbundet med en heterolog promotor, som omfatter et nukleinsyremolekyle med en sekvens, der er mindst 90 % identisk med nukleinsyresekvensen ifølge SEQ ID NO:69, eller som omfatter et nukleinsyremolekyle med en sekvens, der er mindst 90 % identisk med nukleinsyresekvensen ifølge SEQ ID NO:71; hvor den heterologe promotors operative binding til mindst ét polyumættet fedtsyre-biosyntese-polynukleotid ændrer produktionen af ét eller flere polyumættede fedtsyrer via den genmanipulerede celle sammenlignet med en referencecelle, når den genmanipulerede celle og referencecellen dyrkes under sammenlignelige betingelser; og dyrkning af den genmanipulerede Thraustochytrium-ceWe under betingelser og i et tidsrum, der er tilstrækkeligt til at opnå produktion af den eller de polyumættede fedtsyrer.
14. Fremgangsmåde ifølge krav 11 eller 13, hvor den eller de polyumættede fedtsyrer er valgt fra gruppen bestående af alpha-linolensyre, arachidonsyre, docosahexaensyre, docosapentaensyre, eicosapentaensyre, gamma-linolensyre, linolsyre og/eller linolensyre.
15. Fremgangsmåde ifølge krav 14, hvor det mindst ene polyumættede fedtsyre-biosyntese-polynukleotid koder for et enzympolypeptid eller en del af et enzympolypeptidkompleks, som er valgt fra gruppen bestående af fedtsyresynthase (FAS), A5-elongase, A5-desaturase, A4-desaturase og polyketid-PUFA-synthase (PKS).
DK14713925.7T 2013-03-13 2014-03-12 Genmanipulering af mikroorganismer DK2970926T3 (da)

Applications Claiming Priority (2)

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US201361779257P 2013-03-13 2013-03-13
PCT/IB2014/059687 WO2014141098A1 (en) 2013-03-13 2014-03-12 Engineering microorganisms

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US (1) US9873880B2 (da)
EP (1) EP2970926B1 (da)
CN (1) CN105189740B (da)
AU (1) AU2014229307B2 (da)
CA (1) CA2902698A1 (da)
DK (1) DK2970926T3 (da)
ES (1) ES2666895T3 (da)
HK (1) HK1220485A1 (da)
NO (1) NO3022290T3 (da)
PT (1) PT2970926T (da)
WO (1) WO2014141098A1 (da)

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CN105189740B (zh) 2013-03-13 2019-01-15 帝斯曼营养品股份公司 工程化微生物
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CN104293769B (zh) * 2014-10-27 2016-06-01 徐毅 一种来源于绿色巴夫藻的delta8去饱和酶新基因
CN110168071A (zh) * 2016-12-22 2019-08-23 玛拉可再生能源公司 用于使用真核微生物产生富含dha、棕榈酸和蛋白质的生物质的方法
CN111954713A (zh) * 2018-03-05 2020-11-17 科纳根公司 用于产生具有低硫酸化的糖分子的生物体和方法
CN118434873A (zh) * 2021-12-10 2024-08-02 菲科尔生物技术国际公司 原壳异养小球藻中必需长链多不饱和脂质(lcpufa)的异养生产
KR20240141784A (ko) 2022-01-25 2024-09-27 디에스엠 아이피 어셋츠 비.브이. 다중불포화 지방산 생산을 증가시키기 위한 배지 조정 및 영양소 공급 접근법

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CN105189740A (zh) 2015-12-23
ES2666895T3 (es) 2018-05-08
CN105189740B (zh) 2019-01-15
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EP2970926A1 (en) 2016-01-20
AU2014229307A1 (en) 2015-09-03
EP2970926B1 (en) 2018-01-31
PT2970926T (pt) 2018-03-22
US20160040175A1 (en) 2016-02-11
WO2014141098A1 (en) 2014-09-18
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