Methods of producing sesquiterpene compounds

A method of producing one or more sesquiterpene compound derivatives, the method comprising,

• producing one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene by contacting an acyclic farnesol diphosphate (FPP) with a polypeptide having a terpene synthase activity, wherein said polypeptide comprises an isovalencene synthase activity, a spirovetiva-1(10),7(11)-diene synthase activity and/or a valencene synthase activity, wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:
• 1 or comprises the amino acid sequence of SEQ ID NO:
• 1, and
• oxidizing the one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene-to obtain derivatives of the one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene by chemical and/or biochemical synthesis.

A method of producing one or more sesquiterpene compound derivatives, the method comprising,

• producing one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene by contacting an acyclic farnesyl diphosphate (FPP) with a polypeptide having a terpene synthase activity, wherein said polypeptide comprises an isovalencene synthase activity, a spirovetiva-1(10),7(11)-diene synthase activity and/or a valencene synthase activity, wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:
• 1 or comprises the amino acid sequence of SEQ ID NO:1, and
• contacting the one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene with at least one oxidizing enzyme to produce derivatives of the one or more sesquiterpene compounds selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene,
• wherein the oxidizing enzyme is a cytochrome P450 polypeptide having cytochrome P450 oxidizing activity, wherein said cytochrome P450 polypeptide having cytochrome P450 oxidizing activity comprises an amino acid sequence at least 95% sequence identical to the amino acid sequence of SEQ ID NO:
• 7, 10, 12, or 14, or comprises the amino acid sequence of SEQ ID NO:
• 7, 10, 12, or 14.

A method of producing oxygenated sesquiterpene compounds comprising

• oxidizing one or more sesquiterpene compounds to one or more oxygenated sesquiterpene compounds by chemical and/or biochemical synthesis with a cytochrome P450 polypeptide having cytochrome P450 oxidizing activity, wherein the cytochrome P450 polypeptide oxidizes the one or more sesquiterpene compounds accompanied by a cytochrome P450 reductase enzyme for electron transfer to the cytochrome P450 polypeptide,
• wherein the one or more sesquiterpene compounds being oxidized are selected from the group consisting of isovalencene, spirovetiva-1(10),7(11)-diene and valencene, wherein the cytochrome P450 reductase enzyme comprises an amino acid sequence at least 95% sequence identical to the amino acid sequence of SEQ ID NO:15 or comprises the amino acid sequence of SEQ ID NO:15, and
• wherein the cytochrome P450 polypeptide having cytochrome P450 oxidizing activity comprises an amino acid sequence at least 95% sequence identical to the amino acid sequence of SEQ ID NO:
• 7, 10, 12, or 14, or comprises the amino acid sequence of SEQ ID NO:
• 7, 10, 12, or 14.