HRP20200240T1 - Ekspresijsko punjenje za transformaciju eukariotske stanice, postupak za transformaciju eukariotske stanice, genetički modificirani organizam i postupak za proizvodnju biogoriva i/ili biokemikalija i biogoriva koja su proizvedena na taj način - Google Patents

Ekspresijsko punjenje za transformaciju eukariotske stanice, postupak za transformaciju eukariotske stanice, genetički modificirani organizam i postupak za proizvodnju biogoriva i/ili biokemikalija i biogoriva koja su proizvedena na taj način Download PDF

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HRP20200240T1
HRP20200240T1 HRP20200240TT HRP20200240T HRP20200240T1 HR P20200240 T1 HRP20200240 T1 HR P20200240T1 HR P20200240T T HRP20200240T T HR P20200240TT HR P20200240 T HRP20200240 T HR P20200240T HR P20200240 T1 HRP20200240 T1 HR P20200240T1
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seq
microorganism
gene
expression cassette
promoter
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HRP20200240TT
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Leandro Vieira DOS SANTOS
Gonçalo Amarante Guimarães PEREIRA
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Biocelere Agroindustrial Ltda.
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52Genes encoding for enzymes or proenzymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/90Isomerases (5.)
    • C12N9/92Glucose isomerase (5.3.1.5; 5.3.1.9; 5.3.1.18)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00Preparation of oxygen-containing organic compounds
    • C12P7/02Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06Ethanol, i.e. non-beverage
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y503/00Intramolecular oxidoreductases (5.3)
    • C12Y503/01Intramolecular oxidoreductases (5.3) interconverting aldoses and ketoses (5.3.1)
    • C12Y503/01005Xylose isomerase (5.3.1.5)
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02EREDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00Technologies for the production of fuel of non-fossil origin
    • Y02E50/10Biofuels, e.g. bio-diesel

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Claims (15)

1. Ekspresijska kazeta za transformaciju eukariotske stanice naznačena time što sadrži kombinaciju sljedećih ekspresijskih kazeta: a) ekspresijsku kazetu koja sadrži gen koji kodira ksiloza izomerazu sekvence SEQ ID NO: 1, promotor TDH1 nukleotidne sekvence SEQ ID NO: 2 i terminator TDH1 nukleotidne sekvence SEQ ID NO: 3; b) ekspresijsku kazetu koja sadrži promotor ADH1 predstavljen sekvencom SEQ ID NO: 8, gen XKS1 predstavljen sekvencom SEQ ID NO: 9 i terminator ADH1 predstavljen sekvencom SEQ ID NO: 10; c) ekspresijsku kazetu koja sadrži promotor TDH1 nukleotidne sekvence SEQ ID NO: 2, gen TAL1 sekvence SEQ ID NO: 5, gen terminator TDH1 sekvence SEQ ID NO: 3, nakon koga slijedi promotor PGK1 sekvence SEQ ID NO: 6, gen RKI1 (SEQ ID NO: 7) i terminator nukleotidne sekvence SEQ ID NO: 13; i d) ekspresijsku kazetu koja sadrži promotor TDH1 sekvence SEQ ID NO: 02, gen TKL1 sekvence SEQ ID NO: 11, gen koji kodira riboza 5-fosfat epimerazu (SEQ ID NO: 7), terminator TDH1 sekvence SEQ ID NO: 3, nakon koga slijedi promotor PGK1 sekvence SEQ ID NO: 6, gen RPE1 sekvence SEQ ID NO: 12 i terminator PGK1 sekvence SEQ ID NO: 13; pri čemu je navedena ekspresijska kazeta funkcionalna u eukariotskoj stanici.
2. Ekspresijska kazeta, prema patentnom zahtjevu 1, naznačena time što je navedeni promotor(i) konstitutivni ili prirodno inducibilni.
3. Postupak za transformaciju eukariotske stanice, naznačen uvođenjem, u stanicu koja se transformiše, najmanje ekspresione kasete kao što je definirana u bilo kojem od patentnih zahtjeva 1-2.
4. Postupak, prema patentnom zahtjevu 3, dodatno naznačen time što sadrži inaktivaciju ili deleciju GRE3 gena (SEQ ID NO: 14) u genomu navedene eukariotske stanice.
5. Genetički modificirani mikroorganizam, naznačen time što sadrži najmanje ekspresijsku kazetu kao što je definirana u bilo kojem od patentnih zahtjeva 1-2.
6. Mikroorganizam, prema patentnom zahtjevu 5, naznačen time što su jedna ili više navedenih ekspresijskih kazeta prisutne u regiji od 5 tisuća prvih baznih parova brojanih od centromere kako u ushodnom tako i u nishodnom pravcu, i može biti čak samo ushodnom, samo nishodnom ili oba istovremeno.
7. Mikroorganizam, prema patentnom zahtjevu 6, naznačen time što promotorske sekvence, kodirajuće sekvence i terminatorske sekvence ekspresijskih kazeta su stabilne u genomu mikroorganizma i/ili su prisutne u najmanje 5 kopija u genomu mikroorganizma.
8. Mikroorganizam, prema bilo kojem od patentnih zahtjeva 6-7, naznačen time što gen GRE3 (SEQ ID NO: 14) je inaktiviran ili deletiran u/iz njegovog genoma.
9. Mikroorganizam, prema bilo kojem od patentnih zahtjeva 6-8, naznačen time što je to kvasac iz roda izabranog iz grupe koja se sastoji od: Saccharomyces, Scheffersomyces, Spathaspora, Pichia, Candida, Kluyveromyces, Schizosaccharomyces, Brettanomyces, Hansenula i Yarrowia.
10. Mikroorganizam, prema patentnom zahtjevu 9, naznačen time što mikroorganizam je Saccharomyces cerevisiae DSM28739.
11. Genetički modificirani mikroorganizam, naznačen time što je to Saccharomyces cerevisiae DSM28739.
12. Postupak za proizvodnju biogoriva i/ili biokemikalija, naznačen time što sadrži korak kultivacije mikroorganizma kao što je definiran u bilo kojem od patentnih zahtjeva 5-11.
13. Postupak, prema patentnom zahtjevu 12, naznačen time što je prinos postupka najmanje 0.45 grama etanola proizvedenog po gramu ksiloze potrošene od strane mikroorganizma u sintetičkom mediju koji sadrži ksilozu kao izvor ugljika.
14. Postupak, prema patentnom zahtjevu 12 ili 13, naznačen time što volumetrijska produktivnost je najmanje 0.67 grama etanola proizvedenog po litri svakog sata, kada je u sintetičkom mediju koji sadrži ksilozu kao izvor ugljika.
15. Postupak, prema bilo kojem od patentnih zahtjeva 12-14, naznačen time što mikroorganizam je mikroorganizam Saccharomyces cerevisiae DSM28739.
HRP20200240TT 2014-06-12 2020-02-12 Ekspresijsko punjenje za transformaciju eukariotske stanice, postupak za transformaciju eukariotske stanice, genetički modificirani organizam i postupak za proizvodnju biogoriva i/ili biokemikalija i biogoriva koja su proizvedena na taj način HRP20200240T1 (hr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
BR102014014407A BR102014014407A2 (pt) 2014-06-12 2014-06-12 cassete de expressão para transformar célula eucariótica, micro-organismo geneticamente modificado com eficiente consumo de xilose, processo para produção de biocombustíveis e bioquímicos e biocombustível e/ou bioquímico assim produzido
PCT/BR2015/050075 WO2015188244A1 (pt) 2014-06-12 2015-06-12 Cassete de expressão para a transformação de célula eucariótica, processo para a transformação de célula eucariótica, micro-organismo geneticamente modificado, processo de produção de biocombustíveis e/ou bioquímicos e biocombustível e/ou bioquímico assim produzidos
EP15806997.1A EP3165609B1 (en) 2014-06-12 2015-06-12 Expression cartridge for the transformation of a eukaryotic cell, method for transforming a eukaryotic cell, genetically modified organism, and process for producing biofuels and/or biochemicals and biofuels thus produced

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US (1) US10155953B2 (hr)
EP (1) EP3165609B1 (hr)
BR (2) BR102014014407A2 (hr)
DK (1) DK3165609T3 (hr)
ES (1) ES2773639T3 (hr)
HR (1) HRP20200240T1 (hr)
HU (1) HUE048401T2 (hr)
PL (1) PL3165609T3 (hr)
PT (1) PT3165609T (hr)
RS (1) RS59923B1 (hr)
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WO (1) WO2015188244A1 (hr)

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EP3165609A1 (en) 2017-05-10
DK3165609T3 (da) 2020-02-24
US10155953B2 (en) 2018-12-18
BR112016028778B1 (pt) 2023-01-17
PT3165609T (pt) 2020-02-25
RS59923B1 (sr) 2020-03-31
BR112016028778A2 (pt) 2018-01-09
BR102014014407A2 (pt) 2016-04-19
SI3165609T1 (sl) 2020-04-30
US20170114350A1 (en) 2017-04-27
HUE048401T2 (hu) 2020-07-28
WO2015188244A1 (pt) 2015-12-17
EP3165609B1 (en) 2019-11-13
ES2773639T3 (es) 2020-07-13
EP3165609A4 (en) 2017-09-27
PL3165609T3 (pl) 2020-06-01

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