Sequences for fk228 biosynthesis and methods of synthesizing fk228 and fk228 analogs
AssigneeUWM Research Foundation, Inc.
InventorYi-Qiang Cheng
Life of this patent
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Abstract
Polynucleotides encoding the polypeptides involved in biosynthesis of FK 228 and those involved in synthesis of a novel FK228 analog, thailandepsin are disclosed herein. Also provided are methods of making FK228, thailandepsin and analogs of these molecules and methods of using these FK228 analogs. Chromobacterium and Burkholderia gene inactivation mutants are provided. Methods of forming a disulfide bond in a chemical are also disclosed.
Claims21
-28. (canceled)
A method of making FK228, thailandepsin, an FK228 analog, or a thailandepsin analog comprising growing a recombinant cell comprising polynucleotides encoding proteins selected from the group consisting of depA, depB, depC, depD, depE, depF, depG, depH, depI, depJ, depK, depL, depM, depN, tdpA, tdpB, tdpC1, tdpC2, tdpDE1, tdpE2, tdpF, tdpG, tdpH, tdpI, tdpJ, tdpL, and tdpN or a homolog thereof, the polynucleotides operably connected to a promoter, under conditions that allow synthesis of FK228, thailandepsin, an FK228 analog, or a thailandepsin analog.
-37. (canceled)
-44. (canceled)
A method of making an FK228 analog or a thailandepsin analog comprising:
- introducing a polynucleotide into Chromobacterium violaceum No. 968 or Burkholderia thailandensis E264 to produce a recombinant bacterium, wherein the polynucleotide encodes a polypeptide that is a homolog of at least one of DepA, DepB, DepC, DepD, DepE, DepF, DepG, DepH, DepI, DepJ, DepK, DepL, DepM, DepN, TdpA, TdpB, TdpC1, TdpC2, TdpDE1, TdpE2, TdpF, TdpG, TdpH, TdpI, TdpJ, TdpL, and TdpN and the polynucleotide is operably connected to a promoter;
- and
- growing the recombinant bacterium under conditions that allow expression of the polynucleotide and production of the FK228 analog or the thailandepsin analog.
(canceled)
-62. (canceled)
A method of producing an FK228 analog comprising growing Burkholderia thailandensis E264 in medium and partially isolating the FK228 analog from the growth medium.
(canceled)
Dependencies are inferred from the claim text.