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US 10,208,326 B2Granted
Methods and materials for biosynthesis of manoyl oxide
AssigneeEvolva SA
InventorNiels Bjerg Jensen
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Abstract
The invention relates to recombinant microorganisms and methods for producing manoyl oxide.
Claims15
A method of producing a manoyl oxide, comprising:
- growing a recombinant host, comprising:
- (a) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide,
- wherein the GGPPS polypeptide comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:2 or 4;
- (b) a gene encoding a polypeptide catalyzing formation of copal-8-ol diphosphate from qeranylgeranyl diphosphate (GGPP),
- wherein the polypeptide catalyzing formation of copal-8-ol diphosphate from GGPP comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:6;
- and
- (c) a gene encoding a polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate,
- wherein the polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:9 or 11 in a culture medium, under conditions in which the genes are expressed;
- wherein the manoyl oxide is synthesized by the host.
6 more elements
A method for producing a terpenoid, comprising:
- (a) culturing a recombinant host capable of producing a manoyl oxide, comprising:
- (i) a gene encoding a geranylgeranyl diphosphate synthase (GGPPS) polypeptide,
- wherein the GGPPS polypeptide comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:2 or 4;
- (ii) a gene encoding a polypeptide catalyzing formation of copal-8-ol diphosphate from geranylgeranyl diphosphate (GGPP),
- wherein the polypeptide catalyzing formation of copal-8-ol diphosphate from GGPP comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:6;
- and
- (iii) a gene encoding a polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate,
- wherein the polypeptide catalyzing formation of manoyl oxide from copal-8-ol diphosphate comprises a polypeptide having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NOs:9 or 11;
- wherein at least one of the genes is a recombinant gene;
- in a culture medium, under conditions in which the genes are expressed;
- wherein the manoyl oxide is synthesized by the recombinant host;
- (b) isolating the manoyl oxide produced by the recombinant host;
- and
- (c) converting the manoyl oxide into the terpenoid.
11 more elements
Dependencies are inferred from the claim text.