Microbial oils with lowered pour points, dielectric fluids produced therefrom, and related methods

• US 10,167,489 B2
• Filed: 06/10/2016
• Issued: 01/01/2019
• Est. Priority Date: 11/03/2010
• Status: Active Grant

• Current Assignee: Corbion Biotech, Inc. (Corbion NV)
• Original Assignee: Corbion Biotech, Inc. (Corbion NV)
• Inventors: Franklin, Scott, Rakitsky, Walter, Rudenko, George, Zhao, Xinhua, Rodriguez, Felipe Arana, Lu, Wenhua, Wee, Janice
• Primary Examiner(s): McGinty, Douglas J
• Application Number:
  US15/179,253
• Publication Number:
  US 20160376617A1
• Time in Patent Office: 935 Days
• Field of Search: 252578, 336 58, 508112, 435134, 4352523, 4352572, 435 41
• CPC Class Codes: A23D 9/007 characterised by ingredients other than fatty acid triglycerides
• CPC Class Codes: A23D 9/02 characterised by the production or working-up
• CPC Class Codes: C10M 169/041 the additives being macromolecular compounds only
• CPC Class Codes: C11B 1/025 by enzymes or microorganisms, living or dead
• CPC Class Codes: C11B 1/06 by pressing
• CPC Class Codes: C11B 1/10 by extracting
• CPC Class Codes: C11B 3/001 by a combination of two or more of the means hereafter
• CPC Class Codes: C11B 3/006 by extraction
• CPC Class Codes: C11B 3/008 by filtration, e.g. including ultra filtration, dialysis C11B3/005 takes precedence
• CPC Class Codes: C11B 3/06 with bases
• CPC Class Codes: C11B 3/10 by adsorption
• CPC Class Codes: C11B 3/14 with the use of indifferent gases or vapours, e.g. steam
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/2442 Chitinase (3.2.1.14)
• 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 7/6409 Fatty acids
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12Y 301/02014 Oleoyl-[acyl-carrier-protein] hydrolase (3.1.2.14), i.e. ACP-thioesterase
• CPC Class Codes: H01B 3/20 liquids, e.g. oils silicone oils H01B3/46
• CPC Class Codes: H01F 27/105 : Cooling by special liquid or by liquid of particular composition
• CPC Class Codes: H01F 27/321 : using a fluid for insulating purposes only

Abstract

Methods and compositions for the production of dielectric fluids from lipids produced by 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 sucrose transporter, a sucrose invertase, a fructokinase, a polysaccharide-degrading enzyme, a lipid pathway modification enzyme, a fatty acyl-ACP thioesterase, a desaturase, a fatty acyl-CoA/aldehyde reductase, and/or an acyl carrier protein are useful in manufacturing dielectric fluids.

Claims

1. A method of producing a microalgal oil, the method comprising:

a. cultivating a genetically engineered Chlorella or Prototheca cell engineered to ablate or downregulate expression of an endogenous fatty acyl-ACP thioesterase gene until the microbe has at least 10% oil by dry weight;

b. separating the oil from the microbe; and

optionally

c. subjecting the oil to refining, bleaching, deodorizing or degumming to produce RBD microbial oil.

2. The method of claim 1, wherein the endogenous fatty acyl-ACP thioesterase gene is downregulated by an inhibitory RNA.

3. The method of claim 1, wherein the Prototheca or Chlorella cell is of the species Prototheca moriformis or Chlorella protothecoides.

4. The method of claim 1, wherein the Chlorella or Prototheca cell further comprises one or more expressed exogenous genes that encode a sucrose invertase, a fatty acyl-ACP thioesterase or a desaturase.

5. The method of claim 4, wherein the one or more expressed exogenous genes encode(s) an inhibitory RNA that targets an endogenous desaturase.

11. The method of claim 1, wherein the endogenous fatty acyl-ACP thioesterase gene is ablated.

12. The method of claim 11, wherein the Prototheca or Chlorella cell is of the species Prototheca moriformis or Chlorella protothecoides.

13. The method of claim 12, wherein the Chlorella or Prototheca cell further comprises one or more expressed exogenous genes that encode a sucrose invertase, a fatty acyl-ACP thioesterase or a desaturase.

14. The method of claim 13, wherein the one or more expressed exogenous genes encode(s) an inhibitory RNA that targets an endogenous desaturase.

6. A method of producing a microalgal oil, the method comprising:

a. cultivating a genetically engineered Chlorella or Prototheca cell engineered to ablate or downregulate expression of an endogenous fatty acyl-ACP thioesterase gene and engineered to express one or more exogenous genes until the microbe has at least 10% oil by dry weight;

b. separating the oil from the microbe; and

optionally

c. subjecting the oil to refining, bleaching, deodorizing or degumming to produce RBD microbial oil.

7. The method of claim 6, wherein the endogenous fatty acyl-ACP thioesterase gene is downregulated by an inhibitory RNA.

8. The method of claim 6, wherein the one or more expressed exogenous genes encode(s) sucrose invertase, fatty acyl-ACP thioesterase or desaturase.

9. The method of claim 6, wherein the one or more expressed exogenous genes encode(s) an inhibitory RNA that targets an endogenous desaturase.

10. The method of claim 6, wherein the Prototheca or Chlorella is of the species Prototheca moriformis or Chlorella protothecoides.

15. The method of claim 6, wherein the endogenous fatty acyl-ACP thioesterase gene is ablated.

16. The method of claim 15, wherein the one or more expressed exogenous genes encode(s) sucrose invertase, fatty acyl-ACP thioesterase or desaturase.

17. The method of claim 16, wherein the one or more expressed exogenous genes encode(s) an inhibitory RNA that targets an endogenous desaturase.

18. The method of claim 17, wherein the Prototheca or Chlorella is of the species Prototheca moriformis or Chlorella protothecoides.