Yeast strains with reduced fatty alcohol oxidase activity and method for the production of omega-hydroxy fatty acids and dicarboxylic acids

• US 10,640,796 B2
• Filed: 08/13/2018
• Issued: 05/05/2020
• Est. Priority Date: 12/12/2013
• Status: Active Grant

• Current Assignee: Provivi, Inc.
• Original Assignee: Provivi, Inc.
• Inventors: Gatter, Michael, Matthaus, Falk, Barth, Gerold
• Primary Examiner(s): Fronda, Christian L
• Application Number:
  US16/102,213
• Publication Number:
  US 20180371510A1
• Time in Patent Office: 631 Days
• Field of Search: None
• CPC Class Codes: C12N 1/165 Yeast isolates
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12N 9/001 acting on the CH-CH group of donors (1.3)
• CPC Class Codes: C12N 9/90 Isomerases (5.)
• CPC Class Codes: C12P 7/44 Polycarboxylic acids
• CPC Class Codes: C12P 7/6409 Fatty acids
• CPC Class Codes: C12P 7/6427 Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
• CPC Class Codes: C12R 2001/73 Candida lipolytica
• CPC Class Codes: C12Y 101/0302 Long-chain-alcohol oxidase (1.1.3.20)
• CPC Class Codes: C12Y 103/03006 Acyl-CoA oxidase (1.3.3.6)
• CPC Class Codes: C12Y 502/01008 Peptidylprolyl isomerase (5.2.1.8), i.e. cyclophilin

Abstract

The invention relates to various new yeast strains of the type Yarrowia lipolytica as well as relevant methods for the biocatalytic preparation of ω-hydroxy fatty acids or dicarboxylic acids with the aid of these strains, whereby the formation of ω-hydroxy fatty acids or dicarboxylic acids is advantageously increased.

Claims

1. A genetically engineered Yarrowia lipolytica cell, comprising:

a. a disrupted fatty alcohol oxidase encoding FAO1 YALI0B14014g gene; and

b. at least one disrupted acyl-CoA oxidase encoding gene selected from the group consisting of;

POX1, POX2, POX3, POX4, POX5, and POX6,

wherein the engineered Yarrowia lipolytica cell has reduced fatty alcohol oxidase activity relative to a wild type Yarrowia lipolytica cell.

2. The engineered Yarrowia lipolytica cell according to claim 1, wherein the fatty alcohol oxidase encoding FAO1 YALI0B14014g gene is partially deleted.

3. The engineered Yarrowia lipolytica cell according to claim 1, wherein the fatty alcohol oxidase encoding FAO1 YALI0B14014g gene is completely deleted.

4. The engineered Yarrowia lipolytica cell according to claim 1, wherein at least one acyl-CoA oxidase encoding gene selected from the group consisting of:

POX1, POX2, POX3, POX4, POX5, and POX6 is partially deleted.

5. The engineered Yarrowia lipolytica cell according to claim 1, wherein at least one acyl-CoA oxidase encoding gene selected from the group consisting of:

POX1, POX2, POX3, POX4, POX5, and POX6 is completely deleted.

6. The engineered Yarrowia lipolytica cell according to claim 1, having reduced acyl-CoA oxidase activity relative to a wild type Yarrowia lipolytica cell.

7. The engineered Yarrowia lipolytica cell according to claim 1, comprising:

at least one disrupted fatty alcohol dehydrogenase encoding gene selected from the group consisting of;

FADH, ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, and ADH7.

8. The engineered Yarrowia lipolytica cell according to claim 1, wherein at least one fatty alcohol dehydrogenase encoding gene selected from the group consisting of:

FADH, ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, and ADH7 is partially deleted.

9. The engineered Yarrowia lipolytica cell according to claim 1, wherein at least one fatty alcohol dehydrogenase encoding gene selected from the group consisting of:

FADH, ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, and ADH7 is completely deleted.

10. The engineered Yarrowia lipolytica cell according to claim 1, further comprising:

at least one disrupted fatty aldehyde dehydrogenase encoding gene selected from the group consisting of FALDH1, FALDH2, FALDH3 and FALDH4.

11. The engineered Yarrowia lipolytica cell according to claim 1, wherein at least one fatty aldehyde dehydrogenase encoding gene selected from the group consisting of FALDH1, FALDH2, FALDH3 and FALDH4 is partially or completely deleted.

12. The engineered Yarrowia lipolytica cell according to claim 1, having reduced fatty alcohol dehydrogenase activity relative to a wild type Yarrowia lipolytica cell.

13. The engineered Yarrowia lipolytica cell according to claim 1, comprising:

at least one of the following genes disrupted selected from the group consisting of;

phosphatidic acid dephosphohydrolase (PAH1), phospholipid diacylglycerol acyltransferase (LRO1), diacylglycerol acyltransferase (DGA1), and glycerol-3-phosphate acyltransferase (SCT1).

14. The engineered Yarrowia lipolytica cell of claim 13, further comprising:

a disrupted acyl-CoA oxidase encoding gene selected from the group consisting of;

POX1, POX2, POX3, POX4, POX5, and POX6.

15. The engineered Yarrowia lipolytica cell of claim 13, further comprising:

at least one disrupted fatty alcohol dehydrogenase encoding gene selected from the group consisting of;

FADH, ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, and ADH7.

16. The engineered Yarrowia lipolytica cell of claim 13, wherein the disruption of the genes reduces the size of the lipid bodies.

17. The engineered Yarrowia lipolytica cell according to claim 1, comprising:

a disrupted peroxisome biogenesis factor 10 protein (PEX10).

18. The engineered Yarrowia lipolytica cell of claim 17, further comprising:

a disrupted acyl-CoA oxidase encoding gene selected from the group consisting of;

POX1, POX2, POX3, POX4, POX5, and POX6.

19. The engineered Yarrowia lipolytica cell of claim 17, further comprising:

at least one disrupted fatty alcohol dehydrogenase encoding gene selected from the group consisting of;

FADH, ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, and ADH7.

20. A genetically engineered Yarrowia lipolytica cell, comprising:

a disrupted fatty alcohol oxidase encoding FAO1 YALI0B14014g gene.

21. The engineered Yarrowia lipolytica cell of claim 20, further comprising a disrupted peroxisome biogenesis factor 10 protein (PEX10).