Method for producing monoterpene and monoterpinoid compounds and use thereof
In various embodiments, the present disclosure provides a method and enzyme for forming various compounds, such as monoterpenes and monoterpenoid compounds. In a specific example, the present disclosure provides a method for producing one or more of (−)-ipsdienol, (−)-ipsenol, ipsenone, and ipsdienone. The present disclosure also provides methods of using compounds formed from the disclosed method and enzyme.
1. An isolated, synthetic, or recombinant polynucleotide sequence at least 95% homologous to the sequence of SEQ ID NO:1 which encodes a polypeptide having oxidation reduction or oxidoreductase activity.
2. The polynucleotide sequence of claim 1 , wherein the sequence is at least 99% homologous to the sequence of SEQ ID NO:1.
3. A fragment of the polynucleotide sequence of claim 1 , that encodes a polypeptide having oxidation reduction or oxidoreductase activity.
4. A method of producing a polypeptide having oxidation or reduction activity comprising:
introducing the polynucleotide of claim 1 into an isolated host cell;
culturing the host cell;
expressing from the host cell a polypeptide, wherein the polypeptide has oxidation or reduction activity; and
isolating the polypeptide.
5. A method of generating a variant of a polypeptide having oxidation or reduction activity comprising:
providing a template nucleic acid comprising a sequence encoding a polypeptide at least 95% amino acid sequence similarity to the polypeptide encoded by the sequence of SEQ ID NO:1;
modifying, deleting or adding one or more nucleotides in the template sequence, or a combination thereof, to generate a variant of the template nucleic acid, wherein the variant encodes a polypeptide having oxidation or reduction activity; and
determining whether the polypeptide encoded by the variant catalytically oxidizes or reduces a substrate.
6. The method of claim 5 , wherein the substrate comprises one or more of ipsenone, ipsenol, ipsdienone, and ipsdienol.
7. A transgenic host cell produced by inserting the polynucleotide of claim 1 into the host cell.
8. An isolated transgenic host cell produced by inserting a polynucleotide of claim 1 .
9. The isolated, synthetic, or recombinant polynucleotide sequence of claim 1 , wherein the polynucleotide sequence is the sequence of SEQ ID NO:1.