Artemisinic epoxide and methods for producing same
AssigneeThe Regents of the University of California
InventorsJeffrey Allen DIETRICH, Yasuo Yoshikuni, Jay D. Keasling, Michelle Chia-Yu Chang
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Abstract
The present invention provides artemisinic epoxide, and methods of synthesizing artemisinic epoxide in a genetically modified host cell. The present invention further provides methods for producing artemisinin. The present invention further provides variant enzymes that catalyze the oxidation of amorpha-4,11-diene to artemisinic epoxide; nucleic acids encoding the variant enzymes; as well as recombinant vectors and host cells comprising the nucleic acids.
Claims24
A genetically modified host cell capable of producing amorpha-4,11-diene, wherein the genetically modified host cell comprises a heterologous cytochrome P450 enzyme that converts amorpha-4,11-diene into amorpha-4-ene-11,12-epoxide.
A genetically modified host cell that produces isopentenyl pyrophosphate via a mevalonate pathway, wherein the genetically modified host cell comprises:
- a) a heterologous nucleic acid comprising a nucleotide sequence encoding amorphadiene synthase;
- b) a heterologous nucleic acid comprising a nucleotide sequence encoding a cytochrome P450 enzyme that converts amorpha-4,11-diene into amorpha-4-ene-11,12-epoxide, wherein said cytochrome P450 enzyme is soluble in the cytosol of said genetically modified host cell.
A genetically modified Escherichia coli host cell capable of producing amorpha-4,11-diene via a heterologous mevalonate pathway, the genetically modified host cell comprising:
- a) a functionally disabled tryptophanase A gene;
- and
- b) a heterologous nucleic acid comprising a nucleotide sequence encoding a cytochrome P450 enzyme having a substitution at amino acid position 87 from phenylalanine to alanine, compared to the amino acid sequence set forth in SEQ ID NO:1.
Dependencies are inferred from the claim text.