Genetically engineered microorganisms that metabolize xylose

• US 8,846,352 B2
• Filed: 05/04/2012
• Issued: 09/30/2014
• Est. Priority Date: 05/06/2011
• Status: Active Grant

• Current Assignee: Corbion Biotech, Inc. (Corbion NV)
• Original Assignee: Solazyme Inc.
• Inventors: Chua, Penelope, Somanchi, Aravind
• Primary Examiner(s): RAGHU, GANAPATHIRAM
• Application Number:
  US13/464,948
• Publication Number:
  US 20120329109A1
• Time in Patent Office: 879 Days
• Field of Search: 435/134, 435/257.2, 435/69.1, 435/91.1, 435/320.1, 554/227, 536/23.1, 536/23.2, 530/350
• US Class Current: 435/134
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 1/22 Processes using, or culture media containing, cellulose or hydrolysates thereof
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/8246 Non-starch polysaccharides, e.g. cellulose, fructans, levans
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12P 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source ethanol C12P7/10
• 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 7/6463 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: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02T 50/678 Aviation using fuels of non-fossil origin

Abstract

The invention provides methods of cultivating oil-bearing microbes using xylose as a fixed carbon source. Also provided are microorganisms that have been genetically engineered to metabolize xylose as a fixed carbon source allowing them to convert xylose into oils. Particular advantages of the processes provided herein include production of oils rather than alcohols through the microbial fermentation processes utilizing xylose.

Claims

1. A recombinant oleaginous microalga comprising an exogenous gene that encodes a xylulose-5-phosphate translocator, wherein the xylulose-5-phosphate translocator is S106SAD-XPT having SEQ ID NO:

49, GPT-A-XPT having SEQ ID NO;

45, or GPT-F-XPT having SEQ ID NO;

47.

2. The recombinant oleaginous microalga of claim 1, further comprising one or more exogenous genes that encode a xylose transporter protein, a symporter protein, an oxidoreductase pathway protein, or a xylose isomerase pathway protein.

3. The recombinant oleaginous microalga of claim 1, wherein the xylulose-5-phosphate translocator is S106SAD-XPT having SEQ ID NO:

49.

4. The recombinant oleaginous microalga of claim 2, wherein the one or more exogenous genes that encode an oxidoreductase pathway protein is XYL3 having SEQ ID NO:

51.

5. The recombinant oleaginous microalga of claim 2, wherein the microalga is of the genus Prototheca.

6. The recombinant oleaginous microalga of claim 5, wherein the microalga is Prototheca kruegani, Prototheca wickerhamii, Prototheca stagnora, Prototheca moriformis, or Prototheca zopfii.

7. The recombinant oleaginous microalga of claim 1, wherein the xylulose-5-phosphate translocator is GPT-A-XPT having SEQ ID NO:

45.

8. The recombinant oleaginous microalga of claim 1, wherein the xylulose-5-phosphate translocator is GPT-F-XPT having SEQ ID NO:

47.

9. The recombinant oleaginous microalga of claim 2, wherein the one or more exogenous genes that encode a xylose isomerase pathway protein is XylA having SEQ ID NO:

15.

10. The recombinant oleaginous microalga of claim 2, wherein the one or more exogenous genes that encode an oxido-reductase pathway protein is XYL1 having SEQ ID NO:

35.

11. The recombinant oleaginous microalga of claim 2, wherein the one or more exogenous genes that encode an oxido-reductase pathway protein is XYL2 having SEQ ID NO:

50.