DK2066778T3 - Fremstilling af isoprenoider og isoprenoid-prækursorer - Google Patents
Fremstilling af isoprenoider og isoprenoid-prækursorer Download PDFInfo
- Publication number
- DK2066778T3 DK2066778T3 DK07838895.6T DK07838895T DK2066778T3 DK 2066778 T3 DK2066778 T3 DK 2066778T3 DK 07838895 T DK07838895 T DK 07838895T DK 2066778 T3 DK2066778 T3 DK 2066778T3
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- DK
- Denmark
- Prior art keywords
- synthase
- promoter
- host cell
- nucleotide sequence
- mevalonate
- Prior art date
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
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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
- C12N9/88—Lyases (4.)
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Claims (16)
1. Genmodificeret værtscelle, som kan producere en isoprenoid, og som omfatter: a) (i) en kodende region omfattende en nukleotidsekvens, der koder for en me-valonatkinase (MK); en nukleinsyre omfattende en nukleotidsekvens, der koder for et eller flere mevalonat-pathway-enzymer, som ikke er MK, hvor kopiantallet af kodende regioner omfattende nukleotidsekvenser, der koder for MK'en, er højere end kopiantallet af kodende regioner, der koder for det ene eller de flere mevalo-nat-pathway-enzymer, som ikke er MK; og (ii) en heterolog nukleotidsekvens, der koder for en terpensyntase; eller b) (i) en kodende region, der koder for en MK, hvor den MK-kodende region er under styring af en første promoter; (ii) en nukleinsyre omfattende en nukleotidsekvens, der koder for et eller flere mevalonat-pathway-enzymer, som ikke er mevalonatkinase (MK), under styring af en anden promoter, hvor den anden promoter er svagere end den første promoter i værtscellen; og (iii) en heterolog nukleotidsekvens, der koder for en terpensyntase; hvor, i tilfælde af (a) eller (b) ovenfor, niveauet af MK, der produceres i værtscellen, er mindst 50 % højere end niveauet af det ene eller de flere mevalonat-pathway-enzymer, som ikke er MK, og mevalonat-pathway-enzymet, som ikke er MK, er udvalgt blandt acetoacetyl-CoA-thiolase, hydroxymethyl-glutaryl-CoenzymA (HMG-Co)-syntase (HMGS), HMG-CoA-reduktase (HMGR), phosphomevalonatkinase (PMK) og mevalonatpyrophosphatdecar-boxylase (MPD).
2. Værtscelle ifølge krav 1, hvor den MK-kodende region er under styring af en første promoter, og nukleotidsekvensen, der koder for et eller flere mevalonat-pathway-enzymer, som ikke er MK, er under styring af en anden promoter, hvor den anden promoter er svagere end den første promoter i værtscellen.
3. Værtscelle ifølge krav 2, hvor den første promoter er en konstitutivt aktiv promoter.
4. Værtscelle ifølge krav 2, hvor den første promoter er en inducerbar promoter.
5. Værtscelle ifølge krav 2, hvor den første promoter er stærkere end en nativ Escherichia coli Lac-promoter eller en PLac-promoter.
6. Værtscelle ifølge krav 2, hvor den første promoter en en lacllV5-promoter eller en trc-promoter.
7. Værtscelle ifølge krav 1, hvor cellen omfatter mindst to kodende regioner omfattende nukleotidsekvenser, der koder for en MK.
8. Værtscelle ifølge krav 1, hvor kopiantallet af kodende regioner omfattende nukleotidsekvenser, der koder for en MK, er højere end kopiantallet af kodende regioner, der koder for et eller flere mevalonat-pathway-enzymer, som ikke er MK.
9. Værtscelle ifølge et hvilket som helst af de foregående krav, yderligere omfattende en nukleotidsekvens, der koder for en isopentenylpyrophosphat (IPP)-isomerase.
10. Værtscelle ifølge et hvilket som helst af de foregående krav, yderligere omfattende en heterolog nukleinsyre omfattende en nukleotidsekvens, der koder for en prenyltransferase udvalgt blandt en farnesyldiphosphatsyntase, en geranyldiphosphatsyntase og en geranylgeranyldiphosphatsyntase.
11. Værtscelle ifølge et hvilket som helst af de foregående krav, hvor isopre-noiden er en diterpen, en triterpen eller en sesquiterpen.
12. Værtscelle ifølge et hvilket som helst af de foregående krav, hvor et eller flere af de nukleotidsekvenser, der koder for MK'en eller mevalonat-pathway-enzymer, er kodon-optimerede til ekspression i værtscellen.
13. Værtscelle ifølge et hvilket som helst af de foregående krav, hvor terpen-syntasen er udvalgt blandt en amorpha-4,11-dien-syntase; en beta-caryophyllen-syntase; en germacren-A-syntase; en 8-epicedrol-syntase; en valencen-syntase; en (+)-delta-cadinen-syntase; en germacren-C-syntase; a (E)-beta-farnesen-syntase; en casben-syntase; en vetispiradien-syntase; en 5-epi-aristolochen-syntase; en aristolchen-syntase-alpha-humulen-syntase; en (E,E)-alpha-farnesen-syntase; en (-)-beta-pinen-syntase; en gamma-terpinen-syntase; en limonen-cyklase; en linalool-syntase; en 1,8-cineol-syntase; en (+)-sabinen-syntase; en E-alpha-bisabolen-syntase; en (+)-bornyl-diphosphat-syntase; en levopimaradien-syntase; en abietadien-syntase; en isopimaradien-syntase; en (E)-gamma-bisabolen-syntase; en taxadien-syntase; en copalyl-pyrophosphat-syntase; en kauren-syntase; en longifolen-syntase; en gamma-humulen-syntase; en Delta-selinen-syntase; en beta-phellandren-syntase; en limonen-syntase; en myrcen-syntase; en terpinolen-syntase; en (-)-camphen-syntase; en (+)-3-caren-syntase; en syn-copalyl-diphosphat-syntase; en alpha-terpineol-syntase; en syn-pimara-7,15-dien-syntase; en ent-sandaaracopimaradien-syntase; en stemer-13-en-syntase; en E-beta-ocimen; en S-linalool-syntase; en geraniol-syntase; en gamma-terpinen-syntase; en linalool-syntase; en E-beta-ocimen-syntase; en epi-cedrol-syntase; en alpha-zingiberen-syntase; en guaiadien-syntase; en cascarilladien-syntase; en cis-muuroladien-syntase; en aphidicolan-16b-ol-syntase; en elizabethatrien-syntase; en sandalol-syntase; en patchoulol-syntase; en zinzanol-syntase; en cedrol-syntase; en scareol-syntase, copa-lol-syntase; og en manool-syntase.
14. Værtscelle ifølge et hvilket som helst af de foregående krav, hvor værtscellen er en prokaryotisk celle.
15. Værtscelle ifølge et hvilket som helst af kravene 1 til 13, hvor værtscellen er en eukaryotisk celle.
16. Værtscelle ifølge krav 1, hvor værtscelle er en Escherichia coli-værtscelle omfattende: en første ekspressionsplasmid omfattende en nukleotidsekvens, der koder for en mevalonatkinase; en anden ekspressionsplasmid omfattende en nukleotidsekvens, der koder for flere eller flere mevalonat-pathway-enzymer, som ikke er mevalonatkina-se (MK), hvor mevalonat-pathway-enzymet, som ikke er MK, er udvalgt blandt acetoacetyl-CoA-thiolase, hydroxymethylglutaryl-CoenzymA (HMG-Co)-syntase (HMGS), HMG-CoA-reduktase (HMGR), phosphomevalonat-kinase (PMK) og mevalonatpyrophosphatdecarboxylase (MPD), hvor kopiantallet af det andet ekspressionsplasmid er lavere end kopiantallet af det første ekspressionsplasmid; og en heterolog nukleotidsekvens, der koder for en terpensyntase.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US82697006P | 2006-09-26 | 2006-09-26 | |
| PCT/US2007/020790 WO2008039499A2 (en) | 2006-09-26 | 2007-09-25 | Production of isoprenoids and isoprenoid precursors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2066778T3 true DK2066778T3 (da) | 2016-04-18 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK07838895.6T DK2066778T3 (da) | 2006-09-26 | 2007-09-25 | Fremstilling af isoprenoider og isoprenoid-prækursorer |
| DK15196123.2T DK3020799T3 (da) | 2006-09-26 | 2007-09-25 | Fremstilling af isoprenoider og isoprenoid-prækursorer |
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| Application Number | Title | Priority Date | Filing Date |
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| DK15196123.2T DK3020799T3 (da) | 2006-09-26 | 2007-09-25 | Fremstilling af isoprenoider og isoprenoid-prækursorer |
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| BR (1) | BRPI0716954B1 (da) |
| DK (2) | DK2066778T3 (da) |
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| US8927241B2 (en) | 2009-11-10 | 2015-01-06 | Massachusetts Institute Of Technology | Microbial engineering for the production of chemical and pharmaceutical products from the isoprenoid pathway |
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| GB0105924D0 (en) | 2001-03-09 | 2001-04-25 | Microscience Ltd | Promoter |
| US7192751B2 (en) * | 2001-12-06 | 2007-03-20 | The Regents Of The University Of California | Biosynthesis of amorpha-4,11-diene |
| US7172886B2 (en) * | 2001-12-06 | 2007-02-06 | The Regents Of The University Of California | Biosynthesis of isopentenyl pyrophosphate |
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| AU2005327292B2 (en) * | 2004-05-21 | 2010-11-04 | The Regents Of The University Of California | Method for enhancing production of isoprenoid compounds |
| CA2574593C (en) * | 2004-07-27 | 2016-07-05 | The Regents Of The University Of California | Genetically modified host cells and use of same for producing isoprenoid compounds |
| CN101080490B (zh) | 2004-12-14 | 2012-08-15 | 帝斯曼知识产权资产管理有限公司 | 改进的甲羟戊酸激酶 |
| JP2009504138A (ja) * | 2005-07-05 | 2009-02-05 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | イソプレノイド修飾酵素をコードするポリヌクレオチド、およびその使用方法 |
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Also Published As
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| DK3020799T3 (da) | 2020-09-21 |
| EP2066778B1 (en) | 2016-01-27 |
| WO2008039499A3 (en) | 2008-10-16 |
| EP2066778A4 (en) | 2012-03-14 |
| EP3020799A1 (en) | 2016-05-18 |
| BRPI0716954B1 (pt) | 2024-04-30 |
| WO2008039499A2 (en) | 2008-04-03 |
| US8257957B2 (en) | 2012-09-04 |
| EP3020799B1 (en) | 2020-08-26 |
| EP2066778A2 (en) | 2009-06-10 |
| BRPI0716954A2 (pt) | 2013-10-29 |
| US20100112672A1 (en) | 2010-05-06 |
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