DK2764084T3 - Anvendelse af phosphoketolase i fremstillingen af mevalonat, isoprenoid-udgangsstoffer, og isopren - Google Patents

Anvendelse af phosphoketolase i fremstillingen af mevalonat, isoprenoid-udgangsstoffer, og isopren Download PDF

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DK2764084T3
DK2764084T3 DK12775413.3T DK12775413T DK2764084T3 DK 2764084 T3 DK2764084 T3 DK 2764084T3 DK 12775413 T DK12775413 T DK 12775413T DK 2764084 T3 DK2764084 T3 DK 2764084T3
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polypeptide
phosphoketolase
cells
nucleic acid
heterologous nucleic
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DK2764084T4 (da
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Zachary Q Beck
Andrew C Eliot
Caroline M Peres
Dmitrii V Vaviline
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Danisco Us Inc
Goodyear Tire & Rubber
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    • C12P7/00—Preparation of oxygen-containing organic compounds
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    • C12Y203/00—Acyltransferases (2.3)
    • C12Y203/01—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • C12Y203/01008—Phosphate acetyltransferase (2.3.1.8)
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    • C12Y401/00—Carbon-carbon lyases (4.1)
    • C12Y401/02—Aldehyde-lyases (4.1.2)
    • C12Y401/02009—Phosphoketolase (4.1.2.9)
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    • C12Y402/00—Carbon-oxygen lyases (4.2)
    • C12Y402/03—Carbon-oxygen lyases (4.2) acting on phosphates (4.2.3)
    • C12Y402/03027—Isoprene synthase (4.2.3.27)
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    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/03—Intramolecular oxidoreductases (5.3) transposing C=C bonds (5.3.3)
    • C12Y503/03002—Isopentenyl-diphosphate DELTA-isomerase (5.3.3.2)
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    • C12Y205/00—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5)
    • C12Y205/01—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5) transferring alkyl or aryl groups, other than methyl groups (2.5.1)
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    • 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 (20)

1. Rekombinante celler i stand til at producere isopren, hvor cellerne omfatter en eller flere heterologe nukleinsyrer, der koder for et polypeptid med phosphoketolase-aktivitet og (i) en eller flere heterologe nukleinsyrer, der koder for et eller flere polypeptider af den fuldstændige mevalonat (MVA)-vej og (ii) en heterolog nukleinsyre, der koder for et isopren-syntase-polypeptid eller et polypeptid med isopren-syntase-aktivitet, hvor dyrkning af nævnte rekombinante celler i et egnet medie tilvejebringer produktionen af isopren.
2. Rekombinante celler i stand til at producere isoprenoid-udgangsstoffer, hvor cellerne omfatter en eller flere heterologe nukleinsyrer, der koder for et polypeptid med phosphoketolase-aktivitet og en eller flere heterologe nukleinsyrer, der koder for et eller flere polypeptider af den fuldstændige MVA-vej, hvor dyrkning af nævnte rekombinante celler i et egnet medie tilvejebringer produktionen af isoprenoid-udgangsstoffer.
3. Rekombinante celler i stand til at producere isoprenoider, hvor cellerne omfatter en eller flere heterologe nukleinsyrer, der koder for et polypeptid med phosphoketolase-aktivitet og (i) en eller flere heterologe nukleinsyrer, der koder for et eller flere polypeptider af den fuldstændige MVA-vej og (ii) en heterolog nukleinsyre, der koder for et polyprenylpyrophosphat-syntase-polypeptid, hvor dyrkning af nævnte rekombinante celler i et egnet medie tilvejebringer produktionen af isoprenoider.
4. Cellerne ifølge krav 1, 2 eller 3, hvor polypeptidet med phosphoketolase-aktivitet er i stand til at syntetisere glyceraldehyd-3-phosphat og acetylphosphat fra xylulose-5-phosphat.
5. Cellerne ifølge krav 1, 2 eller 3, hvor polypeptidet med phosphoketolase-aktivitet er i stand til at syntetisere erythrose-4-phosphat og acetylphosphat fra fruktose-6-phosphat.
6. Cellerne ifølge krav 1, 2 eller 3, hvor den heterologe nukleinsyre, der koder for et polypeptid med phosphoketolase-aktivitet er fra en organisme valgt fra gruppen bestående af: Bifidobacterium iongum, Enterococcus gaiiiniarum, Clostridium acetobutyiicum, Nostoc punctiforme, Rhodopseudomonas palustris, Pantoea, Mucilaginibacter paludis, Thermobifida fusca, Bifidobacterium breve, Rahnella aquatili, Bifidobacterium animalis, Gardnerella vaginalis, Streptomyces avermitiiis, Lactobacillus plantarum, og Lactobacillus reuteri.
7. Cellerne ifølge krav 1, 2 eller 3, hvor den heterologe nukleinsyre, der koder for et polypeptid med phosphoketolase-aktivitet er fra en organisme valgt fra gruppen bestående af: Bifidobacterium Iongum, Enterococcus gaiiiniarum, og Clostridium acetobutyiicum.
8. Cellerne ifølge krav 1, (a) hvor den heterologe nukleinsyre, der koder for et isopren-syntase-polypeptid eller et polypeptid med isopren-syntase-aktivitet er et plante-isopren-syntase-polypeptid, eller (b) hvor isopren-syntase-polypeptidet eller et polypeptid med isopren-syntase-aktivitet er et polypeptid fra Pueraria eller Populus eller en hybrid Populus alba x Populus trem ula, eller (c) hvor isopren-syntase-polypeptidet eller et polypeptid med isopren-syntase-aktivitet er fra en organisme valgt fra gruppen bestående af Pueraria montana eller Pueraria lobata, Populus tremuloides, Populus alba, Populus nigra, og Populus trichocarpa.
9. Cellerne ifølge krav 1, 2, eller 3, hvor et eller flere polypeptider af den fuldstændige MVA-vej er valgt fra (a) et enzym, der kondenserer to molekyler af acetyl-CoA for at danne acetoacetyl-CoA; (b) et enzym, der kondenserer acetoacetyl-CoA med acetyl-CoA for at danne HMG-CoA (f.eks. HMG-syntase); (c) et enzym, der omdanner HMG-CoA til mevalonat; (d) et enzym, der fosforylerer mevalonat til mevalonat 5-phosphat; (e) et enzym, der omdanner mevalonat 5-phosphat til mevalonat 5-pyrophosphat; og (f) et enzym, der omdanner mevalonat 5-pyrophosphat til isopentenyl pyrophosphat, hvor de rekombinante celler eventuelt yderligere omfatter en eller flere nukleinsyrer, der koder for et eller flere 1-deoxy-D-xylulose 5-phosphat (DXP)- vej-polypeptider.
10. Cellerne ifølge krav 1, 2 eller 3, hvor den ene eller flere nukleinsyrer er placeret under kontrollen af en inducerbar promoter eller en konstitutiv promoter.
11. Cellerne ifølge krav 1, 2 eller 3, hvor den ene eller flere nukleinsyrer er klonet ind i et eller flere multikopi-plasmider.
12. Cellerne ifølge krav 1, 2 eller 3, hvor den ene eller flere nukleinsyrer er integreret i et kromosom af cellerne.
13. Cellerne ifølge krav 1, 2 eller 3, hvor de rekombinante celler er grampositive bakterieceller, gramnegative bakterieceller, svampeceller, filamentøse svampeceller, algeceller eller gærceller.
14. Cellerne ifølge krav 1, 2 eller 3, hvor de rekombinante celler er valgt fra gruppen bestående af Bacillus subtilis, Streptomyces lividans, Streptomyces coelicolor, Streptomyces griseus, Escherichia coli, Pantoea citrea, Trichoderma reesei, Aspergillus oryzae, Aspergillus niger, Saccharomyces cerevisieae og Yarrowia lipolytica.
15. Cellerne ifølge krav 3, (a) hvor isoprenoidet er valgt fra gruppen bestående af et monoterpen, diterpen, triterpen, tetraterpen, sesquiterpen, og polyterpen, eller (b) hvor isoprenoidet er et sesquiterpen, eller (c) hvor isoprenoidet er valgt fra gruppen bestående af abietadien, amorphadien, caren, a-farnesen, β-farnesen, farnesol, geraniol, geranylgeraniol, linalool, limonen, myrcen, nerolidol, ocimen, patchoulol, β-pinen, sabinen, y-terpinen, terpinden og valencen.
16. Cellerne ifølge krav 1, 2 eller 3, hvor de rekombinante celler er yderligere modificeret til (a) at mindske ekspressionen og/eller aktiviteten af et acetat-kinase-gen, eller (b) at forøge ekspressionen og/eller aktiviteten af et phosphat-acetyltransferase-gen.
17. Cellerne ifølge krav 1, 2 eller 3, hvor produktionen af henholdsvis isopren, isoprenoid-udgangsstoffer, eller isoprenoider sker uden en forøgelse i intracellulær acetyl-CoA.
18. Fremgangsmåde til produktion af isopren, isoprenoid-udgangsstoffer eller isoprenoider, hvilken fremgangsmåde omfatter: (a) at dyrke de rekombinante celler ifølge krav 1, 2 eller 3 under betingelser egnede til produktion af isopren, isoprenoid-udgangsstoffer eller isoprenoider og (b) at producere henholdsvis isopren, isoprenoid-udgangsstoffer eller isoprenoider.
19. De rekombinante celler ifølge krav 1, hvor cellerne omfatter en eller flere heterologe nukleinsyrer, der koder for et polypeptid med phosphoketolase-aktivitet og (i) en eller flere nukleinsyrer, der koder for et eller flere polypeptider af den fuldstændige MVA-vej og (ii) en heterolog nukleinsyre, der koder for et isopren-syntase-polypeptid, og hvor dyrkning af nævnte rekombinante celler i et egnet medie tilvejebringer produktionen af isopren.
20. De rekombinante celler ifølge krav 2, hvor isoprenoid-udgangsstoffet er valgt fra gruppen bestående af MVA, isopentenyl-pyrophosphat (IPP), og dimethylallyl-diphosphat (DMAPP).
DK12775413.3T 2011-10-07 2012-10-05 Anvendelse af phosphoketolase i fremstillingen af mevalonat, isoprenoid-udgangsstoffer, og isopren DK2764084T4 (da)

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US201161545083P 2011-10-07 2011-10-07
PCT/US2012/059136 WO2013066568A1 (en) 2011-10-07 2012-10-05 Utilization of phosphoketolase in the production of mevalonate, isoprenoid precursors, and isoprene

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DK2764084T4 DK2764084T4 (da) 2021-12-06

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DK12775413.3T DK2764084T4 (da) 2011-10-07 2012-10-05 Anvendelse af phosphoketolase i fremstillingen af mevalonat, isoprenoid-udgangsstoffer, og isopren

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US (3) US8993305B2 (da)
EP (2) EP3339426B1 (da)
JP (2) JP6420147B2 (da)
CN (1) CN104053765A (da)
AU (1) AU2012332996A1 (da)
BR (1) BR112014008278A2 (da)
CA (1) CA2851265A1 (da)
DK (2) DK3339426T3 (da)
HK (1) HK1201071A1 (da)
MX (1) MX2014004209A (da)
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