US 2017/0121749 A1 - Method Of Producing Terpenes Or Terpenoids - The Lens - Free & Open Patent and Scholarly Search

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US 2017/0121749 A1

Method Of Producing Terpenes Or Terpenoids

Published: May 4, 2017
Family: 8
Cited Works: 0
Cited by: 0
Cites: 0
Sequences: 73
Additional Info: Full text Sequence

Abstract

The present invention relates to a recombinant Deinococcus bacterium exhibiting enhanced 2-C-methyl-D-erythritol 4-phosphate/1-deoxy-D-xylulose 5-phosphate (MEP/DXP) pathway, and its use for producing terpene or terpenoid compounds.

Claims

1-22. (canceled)
A method of producing a terpene or terpenoid comprising (i) culturing a recombinant Deinococcus bacterium that is genetically modified to increase 1-deoxyxylulose 5-phosphate synthase (DXS) activity and/or isopentenyl pyrophosphatase isomerase (IPP isomerase) activity, under conditions suitable to produce the terpene or terpenoid and optionally (ii) recovering the terpene or terpenoid.
The method of claim 23, wherein the recombinant Deinococcus bacterium overexpresses a native, homologous or heterologous idi gene.
The method of claim 23, wherein the recombinant Deinococcus bacterium overexpresses a native, homologous or heterologous dxs gene.
The method of claim 23, wherein the recombinant Deinococcus bacterium expresses an improved DXS enzyme.
The method of claim 26, wherein the improved DXS enzyme is a mutant DXS enzyme comprising a cysteine at position corresponding to position 244 of SEQ ID NO: 52.
The method of claim 26, wherein the recombinant Deinococcus bacterium expresses a gene encoding the R244C mutant of the DXP synthase from D. radiopugnans (SEQ ID NO: 8), a gene encoding the R238C mutant of the DXP synthase from D. yunweiensis (SEQ ID NO: 14) or a gene encoding the R241C mutant of the DXP synthase from D. geothermalis (SEQ ID NO: 56).
The method of claim 23, wherein the recombinant Deinococcus bacterium expresses an improved DXS enzyme and overexpresses a native, homologous or heterologous idi gene.
The method of claim 23, wherein the recombinant Deinococcus bacterium overexpresses a native, homologous or heterologous gene encoding FPP synthase.
The method of claim 30, wherein the gene encoding FPP synthase encodes a polypeptide selected from the group consisting of:
a) a polypeptide comprising all or an active part of the amino acid sequence of SEQ ID NO: 47;

b) a polypeptide having an amino acid sequence having at least 60% sequence identity to SEQ ID NO: 47;

c) a polypeptide encoded by a nucleotide sequence having at least 60% sequence identity to SEQ ID NO: 46; or

d) a polypeptide which is encoded by a nucleic acid sequence that hybridizes under medium/high stringency conditions with (i) the nucleic acid sequence set forth in SEQ ID NO: 46, (ii) its complementary strand, or (iii) a subsequence of (i) or (ii).
The method of claim 30, wherein the FPP synthase further exhibits dimethylallyltransferase activity and/or geranylgeranyl diphosphate synthase activity.
The method of claim 23, wherein, in the recombinant Deinococcus bacterium, at least one gene selected from the group consisting of native, homologous or heterologous dxr, ispD, ispE, ispF, ispG and ispH genes, is overexpressed.
The method of claim 23, wherein the recombinant Deinococcus bacterium further comprises a gene encoding a heterologous terpene synthase.
The method of claim 34, wherein the heterologous terpene synthase is selected from the group consisting of monoterpene synthases, diterpene synthases, triterpene synthases and sesquiterpene synthases.
The method of claim 34, wherein the heterologous terpene synthase is a monoterpene synthase, a cineole synthase, a limonene synthase or a linalool synthase.
The method of claim 34, wherein the heterologous terpene synthase is a sesquiterpene synthase.
The method of claim 23, wherein, in the recombinant Deinococcus bacterium, the gene encoding the lycopene beta-cyclase is inactivated.
The method of claim 23, wherein the terpene or terpenoid is selected from monoterpenes, diterpenes, triterpenes, sesquiterpenes and carotenoids.
The method of claim 23, wherein the terpene or terpenoid is a sesquiterpene.
The method of claim 23, wherein the terpene or terpenoid is a carotenoid.
The method of claim 41, wherein the carotenoid is lycopene or any other carotenoid compound derived from lycopene.
The method of claim 41, wherein the carotenoid is deinoxanthine.
A recombinant Deinococcus bacterium comprising genetic modifications that increase 1-deoxyxylulose 5-phosphate synthase (DXS) activity and/or isopentenyl pyrophosphatase isomerase (IPP isomerase) activity.

Owners (US)

Deinove (Feb 10 2017)
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Applicants

Deinove
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Inventors

Chabot Nicolas
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Castang Sandra
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Bauchart Philippe
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Leonetti Jean-paul
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CPC Classifications

C12N15/52
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C12N9/1022
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C12P23/00
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C12P5/007
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C12Y202/01007
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C12Y205/01001
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C12Y205/0101
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C12Y301/07003
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C12Y402/03
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C12Y503/03002
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C12Y505/01019
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IPC Classifications

C12N15/52
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C12N9/10
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C12P23/00
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C12P5/00
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Document History

Publication: May 4, 2017
US 2017/0121749 A1
Application: Jun 15, 2015
US 201515317994 A
Priority: Jun 15, 2015
EP 2015063335 W
Priority: Jun 13, 2014
EP 14305907 A