Claims
- 1. A recombinant myxobacterial host cell cultivated under controlled oxygen tension conditions, said host cell comprising polyketide synthase (PKS) genes under control of expression control sequences, wherein expression of said PKS genes result in production of polyketides when said host cell is cultivated under conditions that lead to said polyketide production, said host cell also comprising genes encoding active oxygen-sensitive cytochrome P-450 monooxygenase tailoring enzymes.
- 2. The myxobacterial host cell of claim 1 wherein said polyketide is epothilone.
- 3. The myxobacterial host cell of claim 2 wherein said host cell is Sorangium cellulosum strain K111-150.17, and said oxygen sensitive cytochrome P-450 monooxygensase is EpoK epoxidase.
- 4. A method of altering acyltransferase (AT) domain extender unit specificity from malonyl-CoA to methylmalonyl-CoA in epothilone polyketide synthase (PKS) in a host cell, comprising the steps of cultivating said host cell in growth medium under conditions that lead to the production of epothilone, and modulating oxygen tension to excess dissolved oxygen.
- 5. A method of modulating polyketide congener distribution production ratio in a myxobacterial host cell, comprising the steps of cultivating said host cell in growth medium under conditions that lead to the production of polyketides, and modulating oxygen tension during cultivation of said host cells, resulting in shifting production of species of said polyketide congeners.
- 6. The method of claim 5 further comprising the step of supplementing the growth medium with serine.
- 7. The method of claim 5 further comprising the step of supplementing the growth medium with acetate.
- 8. The method of claim 5 further comprising the step of supplementing the growth medium with propionate.
- 9. The method of claim 5 wherein said polyketide is epothilone.
- 10. The method of claim 9 wherein said oxygen tension is modulated to low oxygen tension and said polyketide congener produced is epothilone D.
- 11. The method of claim 5 wherein said host cell is Myxococcus xanthus with an active EpoK+ monooxygenase.
- 12. The method of claim 5 wherein said host cell is Sorangium cellulosum strain Soce90K111-150.17.
- 13. The method of claim 5 wherein said host cell is Myxococcus xanthus strain K111.32.25.
- 14. The method of claim 10 wherein said epothilone D congener produced is obtained under low oxygen tension.
- 15. The method of claim 10 wherein said epothilone D congener is present in up to four fold higher concentration than under excess oxygen tension conditions.
- 16. The method of claim 5 wherein the ratio of epothilone congener production is shifted from epothilone A and B to epothilone C and D congener distribution under depleted oxygen cultivation.
- 17. The method of claim 5 wherein the ratio of epothilone D to C congener production is at least 3 to 1 under excess oxygen conditions.
- 18. The method of claim 17 wherein the myxobacterial host cell is EpoK−.
- 19. The method claim 17 wherein the myxobacterial host cell is Myxococcus xanthus strain K111-40.1.
- 20. The method of claim 19 wherein the oxygen tension is below 50% dissolved oxygen.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims priority to U.S. patent application Serial No. 60/359,821, filed Feb. 25, 2002. The present patent application claims priority and is related to U.S. patent application Ser. No. 09/724,878, filed Nov. 20, 1998, and is related to U.S. Pat. No. 6,303,342 B1, each of which is herein incorporated by reference in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60359821 |
Feb 2002 |
US |