ENHANCED PRODUCTION OF FATTY ACID DERIVATIVES

• US 20150275188A1
• Filed: 03/18/2015
• Published: 10/01/2015
• Est. Priority Date: 05/19/2006
• Status: Abandoned Application

• Current Assignee: Genomatica Incorporated
• Original Assignee: REG Life Sciences, LLC (Genomatica Incorporated)
• Inventors: Hu, Zhihao, Valle, Fernando
• Application Number:
  US14/661,219
• Publication Number:
  US 20150275188A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C07C 69/34 Esters of acyclic saturated polycarboxylic acids having an esterified carboxyl group bound to an acyclic carbon atom
• CPC Class Codes: C07C 69/52 Esters of acyclic unsaturated carboxylic acids having the esterified carboxyl group bound to an acyclic carbon atom
• CPC Class Codes: C07C 69/533 Monocarboxylic acid esters having only one carbon-to-carbon double bond
• CPC Class Codes: C10L 1/02 essentially based on components consisting of carbon, hydrogen, and oxygen only
• CPC Class Codes: C10L 1/026 for compression ignition
• CPC Class Codes: C10L 1/19 Esters ester radical containing compounds; ester ethers; carbonic acid esters C10L1/1802, C10L1/1805, C10L1/1808, C10L1/1811, C10L1/1814, C10L1/1817 take precedence
• CPC Class Codes: C11C 3/003 by esterification of fatty acids with alcohols C11C3/02 takes precedence
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 1/14 Fungi culture of mushrooms A01G18/00; as new plants A01H15/00; Culture media therefor
• CPC Class Codes: C12N 1/16 Yeasts; Culture media therefor
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/001 acting on the CH-CH group of donors (1.3)
• CPC Class Codes: C12N 9/1029 transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/93 Ligases (6)
• CPC Class Codes: C12P 7/62 Carboxylic acid esters
• CPC Class Codes: C12P 7/6409 Fatty acids
• CPC Class Codes: C12P 7/6436 Fatty acid esters
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/649 : Biodiesel, i.e. fatty acid alkyl esters
• CPC Class Codes: C12Y 102/01 : with NAD+ or NADP+ as acceptor (1.2.1)
• CPC Class Codes: C12Y 301/02 : Thioester hydrolases (3.1.2)
• CPC Class Codes: C12Y 602/01 : Acid-Thiol Ligases (6.2.1)
• CPC Class Codes: C12Y 604/01 : Ligases forming carbon-carbon bonds (6.4.1)
• CPC Class Codes: Y02E 50/10 : Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02E 50/30 : Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

Genetically engineered cells and microorganisms are provided that produce fatty alcohols from the fatty acid biosynthetic pathway, as well as methods of their use.

Claims

1-18. -18. (canceled)

19. A genetically modified bacterial cell for production of a fatty alcohol, comprising an exogenous nucleic acid sequence encoding a fatty acid derivative enzyme comprising an acyl-CoA reductase, wherein said bacterial cell converts a carbon source into a fatty alcohol.

20. The bacterial cell of claim 19, wherein said acyl-CoA reductase is encoded by a gene selected from the group consisting of acr1, acrM, BmFAR, mFAR1, and mFAR2.

21. The bacterial cell of claim 20, wherein said acyl-CoA reductase is encoded by said acr1 gene from Acinetobacter baylyi ADP1.

22. The bacterial cell of claim 20, wherein said acyl-CoA reductase is encoded by said acrM gene from Acinetobacter sp. M1.

23. The bacterial cell of claim 20, wherein said acyl-CoA reductase is encoded by said BmFAR gene from Bombyx mori.

24. The bacterial cell of claim 20, wherein said acyl-CoA reductase is encoded by said mFAR1 gene from Mus musculus.

25. The bacterial cell of claim 20, wherein said acyl-CoA reductase is encoded by said mFAR2 gene from Mus musculus.

26. The bacterial cell of claim 19, wherein said cell is further genetically modified to express an exogenous nucleic acid sequence encoding an acyl-CoA synthase.

27. The bacterial cell of claim 26, wherein said acyl-CoA synthase is encoded by a gene selected from the group consisting of fadD, fadDD35, yhfL, fadD1, fadD2, fadDD22, fadK, and faa3p.

28. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadD gene from Escherichia coli.

29. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadDD35 gene from Mycobacterium tuberculosis HR7Rv.

30. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said yhfL gene from Bacillus subtilis.

31. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadD1 gene from Pseudomonas aeruginosa.

32. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadD2 gene from Pseudomonas aeruginosa PAO1.

33. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadDD22 gene from Mycobacterium tuberculosis HR7Rv.

34. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said fadK gene from Escherichia coli.

35. The bacterial cell of claim 27, wherein said acyl-CoA synthase is encoded by said faa3p gene from Saccharomyces cerevisiae.

36. The bacterial cell of claim 26, wherein said cell is further genetically modified to express an exogenous nucleic acid sequence encoding a thioesterase.

37. The bacterial cell of claim 36, wherein said thioesterase is encoded by a gene selected from the group consisting of tesA, ′

tesA, tesB, fatB, fatB1, fatB2, fatB3, fatA, and fatA1.

38. The bacterial cell of claim 37, wherein said thioestrase is encoded by said tesA gene from Escherichia coli.

39. The bacterial cell of claim 37, wherein said thioestrase is encoded by said ′

tesA gene from Escherichia coli.

40. The bacterial cell of claim 37, wherein said thioestrase is encoded by said tesB gene from Escherichia coli.

41. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatB gene from Cinnamonum camphorum, Umbellularia californica, or Arabidopsis thaliana.

42. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatB1 gene from Umbellularia californica.

43. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatB2 gene from Cuphea hookeriana.

44. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatB3 gene from Cuphea hookeriana.

45. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatA gene from Arabidopsis thaliana, Bradyrhiizobium japonicum, or Cuphea hookeriana.

46. The bacterial cell of claim 37, wherein said thioestrase is encoded by said fatA1 gene from Helianthus annus.

47. The bacterial cell of claim 36, wherein said cell is further genetically modified to express an exogenous nucleic acid sequence encoding an acetyl-CoA carboxylase.

48. The bacterial cell of claim 47, wherein said acetyl-CoA carboxylase is encoded by a gene selected from the group consisting of Escherichia coli, Desulfovibrio desulfuricans, Erwinia (micrococcus) amylovora, Kineococcus radiotolerans SRS30216, Rhodospirillum rubrum, and Vibrio furnissii.

49. The bacterial cell of claim 19, wherein said bacterial cell is an Escherichia coli cell.

50. The bacterial cell of claim 19, wherein the fatty alcohol is released into a fermentation media.

51. The bacterial cell of claim 50, wherein the fermentation media comprises one or more of C8, C10, C12, C14, C16 and C18 fatty alcohols.

52. The bacterial cell of claim 50, wherein the fermentation media comprises unsaturated fatty alcohols.

53. The bacterial cell of claim 50, wherein the fermentation media comprises saturated fatty alcohols.

54. The bacterial cell of claim 50, wherein the fermentation media comprises straight chain fatty alcohols.

55. The bacterial cell of claim 50, wherein the fermentation media comprises branched chain fatty alcohols.

56. A genetically modified bacterial cell for production of a fatty alcohol, comprising an exogenous nucleic acid sequence encoding a fatty acid derivative enzyme comprising an acyl-CoA reductase and an acyl-CoA synthase, wherein said bacterial cell converts a carbon source into a fatty alcohol.

57. The bacterial cell of claim 56, wherein said acyl-CoA reductase is encoded by an acr1 gene from Acinetobacter baylyi ADP1, and wherein said acyl-CoA synthase is encoded by a fadD gene from Escherichia coli.

58. The bacterial cell of claim 56, wherein said cell is further genetically modified to express an exogenous nucleic acid sequence encoding a thioesterase.

59. The bacterial cell of claim 58, wherein said thioesterase is encoded by a ′

tesA gene from Escherichia coli.

60. A genetically modified bacterial cell for production of a fatty alcohol, comprising an exogenous nucleic acid sequence encoding a fatty acid derivative enzyme comprising an acyl-CoA reductase, an acyl-CoA synthase, and a thioesterase, wherein said bacterial cell converts a carbon source into a fatty alcohol.

61. The bacterial cell of claim 60, wherein said acyl-CoA reductase is encoded by an acr1 gene from Acinetobacter baylyi ADP1, wherein said acyl-CoA synthase is encoded by a fadD gene from Escherichia coli, and wherein said thioesterase is encoded by a ′

tesA gene from Escherichia coli.