Tailored oils produced from recombinant oleaginous microorganisms

• US 10,100,341 B2
• Filed: 12/18/2015
• Issued: 10/16/2018
• Est. Priority Date: 02/02/2011
• Status: Active Grant

• Current Assignee: Corbion Biotech, Inc. (Corbion NV)
• Original Assignee: Corbion Biotech, Inc. (Corbion NV)
• Inventors: Franklin, Scott, Somanchi, Aravind, Wee, Janice, Rudenko, George, Moseley, Jeffrey L., Rakitsky, Walter, Zhao, Xinhua, Bhat, Riyaz
• Primary Examiner(s): Keogh, Matthew R
• Application Number:
  US14/974,983
• Publication Number:
  US 20160186219A1
• Time in Patent Office: 1,033 Days
• Field of Search: None
• 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 an oil comprising triacylglycerides, or a product produced from the oil, the method comprising:

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

-ketoacyl-ACP synthase I enzyme via homologous recombination, wherein the homologous recombination interrupts or replaces a gene encoding the β

-ketoacyl-ACP synthase I enzyme; and

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

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

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

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

-ketoacyl-ACP synthase I with a nucleic acid encoding a selectable marker, an invertase, a β

-ketoacyl-ACP synthase, a thioesterase, a desaturase, or a hydroxylase.

3. The method of claim 2, wherein the selectable marker is a galactosidase, a hydroxymethylpyrimidine phosphate synthase, an aminoglycoside 3′

-phosphotransferase, or a hygromycin phosphotransferase.

4. The method of claim 1, wherein the microalga is an obligate heterotroph.

5. The method of claim 4, wherein the microalga is Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis, Prototheca moriformis, or Prototheca zopfii.

6. The method of claim 5, wherein the microalga is Prototheca moriformis.