Claims
- 1. A method for producing an isoprenoid compound, said method comprising the steps of:
(a) culturing a host cell under conditions suitable for expressing a first recombinant protein comprising an isoprenoid synthase and a second recombinant protein comprising a cytochrome P450 polypeptide, wherein said first and second recombinant protein catalyze the formation of an isoprenoid compound not normally produced by said host cell; and (b) recovering said isoprenoid compound.
- 2. The method of claim 1, wherein said cytochrome P450 polypeptide is selected from the group consisting of a polypeptide comprising the amino acid sequence of SEQ ID NO: 1, a polypeptide comprising the amino acid sequence of SEQ ID NO: 3, a polypeptide comprising the amino acid sequence of SEQ ID NO: 5, and a polypeptide comprising the amino acid sequence of SEQ. ID NO: 7:
- 3. The method of claim 1, wherein said host cell further comprises a third recombinant protein selected from the group consisting of an acetyltransferase, a methyltransferase, and a fatty acyltransferase.
- 4. The method of claim 1, wherein said host cell expresses an endogenous or recombinant cytochrome reductase.
- 5. The method of claim 1, wherein said host cell is a yeast cell, a bacterial cell, an insect cell, or a plant cell.
- 6. A host cell expressing a recombinant isoprenoid synthase and a recombinant cytochrome P450 polypeptide.
- 7. The host cell of claim 6, wherein said host cell further expresses a recombinant acetyltransferase, a recombinant methyltransferase, or a recombinant fatty acyltransferase.
- 8. The host cell of claim 6, wherein said host cell expresses an endogenous or recombinant cytochrome reductase.
- 9. The host cell of claim 6, wherein said host cell is selected from the group consisting of a yeast cell, a bacterial cell, an insect cell, and a plant cell.
- 10. An isolated cytochrome P450 polypeptide having 80% identity to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3.
- 11. The polypeptide of claim 10, said polypeptide comprising the amino acid sequence of SEQ ID NO: 1.
- 12. The polypeptide of claim 10, said polypeptide comprising the amino acid sequence of SEQ ID NO: 3.
- 13. An isolated cytochrome P450 polypeptide having 85% identity to the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 7.
- 14. The polypeptide of claim 13, said polypeptide comprising the amino acid sequence of SEQ ID NO: 5.
- 15. The polypeptide of claim 13, said polypeptide comprising the amino acid sequence of SEQ ID NO: 7.
- 16. An isolated cytochrome P450 polypeptide having 97% identity to the amino acid sequence of SEQ ID NO: 11.
- 17. The polypeptide of claim 16, said polypeptide comprising the amino acid sequence of SEQ ID NO: 11.
- 18. An isolated cytochrome P450 nucleic acid molecule which encodes a polypeptide having 80% identity to the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 11.
- 19. The isolated nucleic acid molecule of claim 18, wherein said nucleic acid molecule comprises SEQ ID NO: 1.
- 20. The isolated nucleic acid molecule of claim 18, wherein said nucleic acid molecule comprises SEQ ID NO: 3.
- 21. The isolated nucleic acid molecule of claim 18, wherein said nucleic acid molecule comprises SEQ ID NO: 11.
- 22. An isolated cytochrome P450 nucleic acid molecule which encodes a polypeptide having 85% identity to the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 7.
- 23. The isolated nucleic acid molecule of claim 22, wherein said nucleic acid molecule comprises SEQ ID NO: 5.
- 24. The isolated nucleic acid molecule of claim 22, wherein said nucleic acid molecule comprises SEQ ID NO: 7.
- 25. An isolated nucleic acid molecule that specifically hybridizes under highly stringent conditions to any one of the complement of the sequence described in SEQ ID NOS: 2, 4, 6, 8, or 12, wherein said nucleic acid encodes a cytochrome P450 polypeptide.
- 26. A vector comprising an isolated nucleic acid molecule selected from the group consisting of the isolated nucleic acid molecule of any one of claims 18-21 an 22-24.
- 27. A host cell comprising an isolated nucleic acid molecule selected from the group consisting of the isolated nucleic acid molecule of any one of claims 18-21 and 22-24.
- 28. A plant or plant component comprising an isolated nucleic acid molecule selected from the group consisting of the isolated nucleic acid molecule of any one of claims 18-21 and 22-24.
- 29. A process for conferring disease resistance on a plant or plant component, said method comprising the steps of:
(a) introducing into a plant cell a transgene comprising a nucleic acid molecule selected from the group consisting of the isolated nucleic acid molecule of any one of claims 18-21 and 22-24 operably linked to a promoter functional in said plant cell to yield a transformed plant cell; and (b) regenerating a plant or plant component from said transformed plant cell, wherein the nucleic acid molecule is expressed in said plant or plant component, thereby conferring disease resistance on said plant or plant component.
- 30. A method of producing a cytochrome P450 polypeptide in a host cell, said method comprising the steps of:
(a) providing a host cell of claim 27;(b) culturing said host cell under conditions for expressing said isolated nucleic acid molecule encoding said cytochrome P450 polypeptide; and (c) recovering said cytochrome P450 enzyme.
- 31. A method for producing an altered compound, said method comprising the step of contacting said compound with the isolated polypeptide of any one of claims 10-15 under conditions allowing for the hydroxylation, oxidation, demethylation, methylation, or any combination of said enzymatic activity of said compound and recovering said altered compound.
- 32. A hydroxylating agent comprising the isolated polypeptide of any one of claims 10-15.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit of U.S. provisional application Ser. Nos. 60/274,421 and 60/275,597, filed on Mar. 9, 2001 and Mar. 13, 2001, respectively, both of which are hereby incorporated by reference.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60274421 |
Mar 2001 |
US |
|
60275597 |
Mar 2001 |
US |