Tailored oils produced from recombinant heterotrophic microorganisms

• US 8,592,188 B2
• Filed: 05/27/2011
• Issued: 11/26/2013
• Est. Priority Date: 05/28/2010
• Status: Active Grant

• Current Assignee: Terravia Holdings, Inc. (Corbion NV)
• Original Assignee: Solazyme Inc.
• Inventors: Franklin, Scott, Somanchi, Aravind, Wee, Janice, Rudenko, George, Moseley, Jeffrey L., Rakitsky, Walt
• Primary Examiner(s): KAM, CHIH MIN
• Application Number:
  US13/118,365
• Publication Number:
  US 20110294174A1
• Time in Patent Office: 914 Days
• Field of Search: 435/134, 435/41, 435/252.3, 435/254.2, 435/257.2
• US Class Current: 435/134
• CPC Class Codes: A23D 7/00 Edible oil or fat compositions containing an aqueous phase, e.g. margarines
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: C11B 1/00 Production of fats or fatty oils from raw materials
• CPC Class Codes: C11C 3/00 Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom sulfonated fats or oils C07C309/62; factice C08H; drying oils C09F
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/2431 Beta-fructofuranosidase (3.2.1.26), i.e. invertase
• CPC Class Codes: C12N 9/2465 acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12P 7/64 Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
• CPC Class Codes: C12P 7/6445 Glycerides
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12Y 114/19001 Stearoyl-CoA 9-desaturase (1.14.19.1), i.e. DELTA9-desaturase
• CPC Class Codes: C12Y 301/02 Thioester hydrolases (3.1.2)
• CPC Class Codes: C12Y 301/02014 Oleoyl-[acyl-carrier-protein] hydrolase (3.1.2.14), i.e. ACP-thioesterase
• CPC Class Codes: C12Y 302/01022 Alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12Y 302/01026 Beta-fructofuranosidase (3.2.1.26), i.e. invertase
• CPC Class Codes: Y02E 50/10 : Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02P 30/20 : using bio-feedstock
• CPC Class Codes: Y02T 50/678 : Aviation using fuels of non-fossil origin

Abstract

Methods and compositions for the production of oil, fuels, oleochemicals, and other compounds in recombinant microorganisms are provided, including oil-bearing microorganisms and methods of low cost cultivation of such microorganisms. Microalgal cells containing exogenous genes encoding, for example, a lipase, a sucrose transporter, a sucrose invertase, a fructokinase, a polysaccharide-degrading enzyme, a keto acyl-ACP synthase enzyme, a fatty acyl-ACP thioesterase, a fatty acyl-CoA/aldehyde reductase, a fatty acyl-CoA reductase, a fatty aldehyde reductase, a fatty aldehyde decarbonylase, and/or an acyl carrier protein are useful in manufacturing transportation fuels such as renewable diesel, biodiesel, and renewable jet fuel, as well as oleochemicals such as functional fluids, surfactants, soaps and lubricants.

Claims

1. An oil composition produced by cultivating an oleaginous microbe and recovering the oil composition from the microbe, the oil composition comprising triglycerides and sterols produced by the microbe, wherein the fatty acid profile of the triglyceride oil composition comprises at least 60% C18:

1, and less than 7% C18;

2, wherein the triglyceride oil composition is produced by a recombinant oleaginous microbe comprising an exogenous gene, and optionally wherein an endogenous desaturase of the oleaginous microbe has been inactivated or mutated to have less enzymatic activity.

2. The oil composition of claim 1, wherein the fatty acid profile of the triglyceride oil composition comprises at least 70% C18:

1.

3. The oil composition of claim 2, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1.

4. The oil composition of claim 3, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1.

5. The oil composition of claim 1, wherein the fatty acid profile of the triglyceride oil composition comprises less than 5% C18:

2.

6. The oil composition of claim 5, wherein the fatty acid profile of the triglyceride oil composition comprises less than 3% C18:

2.

7. The oil composition of claim 6, wherein the fatty acid profile of the triglyceride oil composition comprises less than 2% C18:

2.

8. The oil composition of claim 1, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1 and less than 5% C18;

2.

9. The oil composition of claim 8, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1 and less than 3% C18;

2.

10. The oil composition of claim 9, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1 and less than 3% C18;

2.

11. The oil composition of claim 10, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1 and less than 2% C18;

2.

12. An oil composition produced by cultivating an oleaginous microalgae and recovering the oil composition from the microalgae, the oil composition comprising triglycerides and sterols produced by the microalgae, wherein the fatty acid profile of the triglyceride oil composition comprises at least 60% C18:

1, and less than 7% C18;

2, wherein the triglyceride oil composition is produced by a recombinant oleaginous microalgae comprising an exogenous gene, and optionally wherein an endogenous desaturase of the oleaginous microalgae has been inactivated or mutated to have less enzymatic activity.

13. The oil composition of claim 12, wherein the microalgae is Prototheca.

14. The oil composition of claim 13, wherein the microalgae is Prototheca moriformis.

15. The oil composition of claim 13, wherein the fatty acid profile of the triglyceride oil composition comprises at least 70% C18:

1.

16. The oil composition of claim 15, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1.

17. The oil composition of claim 16, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1.

18. The oil composition of claim 13, wherein the fatty acid profile of the triglyceride oil composition comprises less than 5% C18:

2.

19. The oil composition of claim 18, wherein the fatty acid profile of the triglyceride oil composition comprises less than 3% C18:

2.

20. The oil composition of claim 19, wherein the fatty acid profile of the triglyceride oil composition comprises less than 2% C18:

2.

21. The oil composition of claim 13, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1 and less than 5% C18;

2.

22. The oil composition of claim 21, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1 and less than 3% C18;

2.

23. The oil composition of claim 22, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1 and less than 3% C18;

2.

24. The oil composition of claim 23, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% C18:

1 and less than 2% C18;

2.

25. The oil composition of claim 13, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% C18:

1 and less than 1% C18;

2.

26. The oil of claim 12, wherein the microalgae comprises 23S rRNA having at least 85% sequence identity to SEQ ID NO:

15.

27. The oil of claim 12, wherein the microalgae comprises 23S rRNA having at least 90% sequence identity to SEQ ID NO:

15.

28. The oil of claim 12, wherein the microalgae comprises 23S rRNA having at least 95% sequence identity to SEQ ID NO:

15.

29. An oil composition, produced by cultivating an oleaginous microbe and recovering the oil composition from the microbe, the oil composition comprising triglycerides and sterols produced by the microbe, wherein the fatty acid profile of the triglyceride oil composition comprises at least 50% saturated fatty acids, wherein the triglyceride oil composition is produced by a recombinant oleaginous microbe comprising an exogenous gene, and optionally wherein an endogenous desaturase of the oleaginous microbe has been inactivated or mutated to have less enzymatic activity.

30. The oil composition of claim 29, wherein the fatty acid profile of the triglyceride oil composition comprises at least 70% saturated fatty acids.

31. The oil composition of claim 30, wherein the fatty acid profile of the triglyceride oil composition comprises at least 80% saturated fatty acids.

32. The oil composition of claim 31, wherein the fatty acid profile of the triglyceride oil composition comprises at least 90% saturated fatty acids.

33. The oil composition of claim 32, wherein the saturated fatty acids comprie at least 30% C12:

0.

34. The oil composition of claim 33, wherein the saturated fatty acids comprise at least 40% C12:

0.

35. The oil composition of claim 34, wherein the saturated fatty acids comprise at least 50% C12:

0.

36. The oil of claim 29, wherein the microbe is a microalgae.

37. The oil of claim 36, wherein the microalgae comprises 23S rRNA having at least 85% sequence identity to SEQ ID NO:

15.

38. The oil of claim 36, wherein the microalgae comprises 23S rRNA having at least 90% sequence identity to SEQ ID NO:

15.

39. The oil of claim 36, wherein the microalgae comprises 23S rRNA having at least 95% sequence identity to SEQ ID NO:

15.