Tailored oils produced from recombinant oleaginous microorganisms

• US 9,249,436 B2
• Filed: 02/02/2012
• Issued: 02/02/2016
• Est. Priority Date: 02/02/2011
• Status: Active Grant

• Current Assignee: Corbion Biotech, Inc. (Corbion NV)
• Original Assignee: Solazyme Inc.
• Inventors: Franklin, Scott, Somanchi, Aravind, Wee, Janice, Rudenko, George, Moseley, Jeffrey L., Rakitsky, Walt, Zhao, Xinhua, Bhat, Riyaz
• Primary Examiner(s): Page, Brent T
• Assistant Examiner(s): KEOGH, MATTHEW R
• Application Number:
  US13/365,253
• Publication Number:
  US 20120203018A1
• Time in Patent Office: 1,461 Days
• Field of Search: None
• US Class Current: 1/1
• 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: C10L 1/02 essentially based on components consisting of carbon, hydrogen, and oxygen only
• CPC Class Codes: C10L 2200/0476 Biodiesel, i.e. defined lower alkyl esters of fatty acids first generation biodiesel
• CPC Class Codes: C10L 2290/26 Composting, fermenting or anaerobic digestion fuel components or materials from which fuels are prepared
• CPC Class Codes: C10L 2290/30 Pressing, compressing or compacting
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• 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: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

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 acid hydroxylase, a desaturase enzyme, 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. A method for producing a natural oil comprising triacylglycerides, or a product produced from the natural oil, the method comprising:

cultivating a cell of a recombinant microalga of the genera Prototheca and Chlorella, the cell comprising recombinant nucleic acids operable to decrease or eliminate the expression of a gene encoding a β

-ketoacyl-ACP synthase II, enzyme via homologous recombination, and express an exogenous acyl-ACP thioesterase; and

recovering the natural oil from the cell, and optionally further processing the natural oil to produce a food, fuel, or chemical product,

wherein the natural oil has an altered fatty acid profile due to the recombinant nucleic acids.

2. The method of claim 1, wherein the decrease or eliminatation of the expression of the β

-ketoacyl-ACP synthase II is due to the interruption or replacement of the gene encoding the β

-ketoacyl-ACP synthase II with a gene encoding an active acyl-ACP thioesterase.

3. The method of claim 1, wherein the recombinant cell further comprises an exogenous gene encoding an oleate 12-hydroxylase, so as to synthesize ricinoleic acid.

11. The method of claim 1, wherein the microalga is selected from Prototheca wicerhamii, Prototheca stagnora, Pototheca portoricensis, Prototheca moriformis, Prototheca zopfii.

12. The method of claim 11 that is Prototheca moriformis.

13. The method of claim 1, wherein the microalga is selected from Chlorella kessleri, Cllorella luteoviridis Chlorella protothecoides, or Chlorella vulgaris.

14. The method of claim 13 that is Chlorella protothecoides.

4. A recombinant microalgal cell of the genera Prototheca or Chlorella comprising recombinant nucleic operable to decrease or eliminate the expression of a gene encoding a β

-ketoacyl-ACP synthase II enzyme via homologous recombination, and express an exogenous acyl-ACP thioesterase.

5. The cell of claim 4, wherein the cell synthesizes fatty acids through a type II fatty acid biosynthesis pathway.

6. The cell of claim 4, wherein the microalga is an obligate heterotroph.

7. The cell of claim 4, wherein the microalga is selected from Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis, Prototheca moriformis, or Prototheca zopfii.

8. The cell of claim 7 that is Prototheca moriformis.

9. The cell of claim 4, wherein the microalga is selected from Chlorella kessleri, Chlorella luteoviridis Chlorella protothecoides, Chlorella vulgaris.

10. The cell of claim 9 that is Chlorella protothecoides.