Engineering microbes and metabolic pathways for the production of ethylene glycol

Patent metadata

Field	Value
Publication number	US-9994876-B2
Application number	US-201514864493-A
Country	US
Kind code	B2
Filing date	Sep 24, 2015
Priority date	Feb 23, 2012
Publication date	Jun 12, 2018
Grant date	Jun 12, 2018

Abstract

The invention relates to recombinant cells and their use in the production of ethylene glycol.

First claim

What is claimed is:

1. A bacterial cell engineered to produce glycolaldehyde from xylose, wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of, xylulokinase relative to a wild-type cell, the cell has reduced or eliminated activity or reduced or eliminated expression of aldehyde dehydrogenase A relative to a wild-type cell, and the cell recombinantly expresses an enzyme that interconverts xylulose and ribulose.

2. The cell of claim 1 , wherein the enzyme that interconverts xylulose and ribulose is D-tagatose 3-epimerase.

3. The cell of claim 2 , wherein the D-tagatose 3-epimerase is encoded by a dte gene obtained from Pseudomonas cichorii.

4. The cell of claim 1 , wherein the cell further recombinantly expresses D-ribulokinase.

5. The cell of claim 4 , wherein the D-ribulokinase is encoded by a fucK gene obtained from Escherichia coli.

6. The cell of claim 4 , wherein the cell further recombinantly expresses D-ribulose-phosphate aldolase.

7. The cell of claim 6 , wherein the cell further has reduced or eliminated activity or reduced or eliminated expression of L-ribulokinase relative to a wild-type cell.

8. The cell of claim 7 , wherein the cell further recombinantly expresses ATP:L-xylulose 1-phosphotransferase, L-xylulose-1-phosphate aldolase and a glycolaldehyde reductase.

9. The cell of claim 1 , wherein the cell is an Escherichia coli cell.

10. The cell of claim 6 , wherein the D-ribulose-phosphate aldolase is encoded by a fucA gene obtained from Escherichia coli.

11. The cell of claim 6 , wherein the cell is an Escherichia coli cell and the xylulokinase is encoded by a xylB gene.

12. The cell of claim 6 , wherein the cell further recombinantly expresses a glycolaldehyde reductase.

13. The cell of claim 12 , wherein the glycolaldehyde reductase is encoded by a fucO gene obtained from Escherichia coli.

14. The cell of claim 1 , wherein the cell comprises a deletion of a gene encoding aldehyde dehydrogenase A and/or a deletion of a gene encoding xylulokinase.

15. The cell of claim 14 , wherein the cell is an Escherichia coli cell and the aldehyde dehydrogenase A is encoded by an aldA gene and/or the xylulokinase is encoded by a xylB gene.

16. A method of producing ethylene glycol, comprising culturing the cell of claim 1 in the presence of xylose under conditions that result in the production of ethylene glycol.

17. A method of producing glycolaldehyde, comprising culturing the cell of claim 6 in the presence of xylose under conditions that result in the production of glycoaldehyde.

18. An Escherichia coli cell engineered to produce glycolaldehyde, wherein the cell comprises a deletion of a xylB gene and recombinantly expresses a dte gene obtained from Pseudomonas cichorii , a fucK gene obtained from Escherichia coli , and a fucA gene obtained from Escherichia coli.

19. A method of producing glycolaldehyde, comprising culturing the cell of claim 18 in the presence of xylose under conditions that result in the production of glycoaldehyde.

20. An Escherichia coli cell engineered to produce ethylene glycol, wherein the cell comprises a deletion of a xylB gene and an aldA gene, and recombinantly expresses a dte gene obtained from Pseudomonas cichorii , a fucK gene obtained from Escherichia coli , a fucA gene obtained from Escherichia coli , and a fucO gene obtained from Escherichia coli.

21. A method of producing ethylene glycol, comprising culturing the cell of claim 20 in the presence of xylose under conditions that result in the production of ethylene glycol.

22. An engineered bacterial cell that comprises a deletion of a gene encoding xylulokinase and a deletion of a gene encoding aldehyde dehydrogenase A.

23. The cell of claim 22 , wherein the cell is an Escherichia coli cell.

24. The cell of claim 22 that further comprises a gene encoding an enzyme that interconverts xylulose and ribulose.

25. The cell of claim 24 , wherein the enzyme that interconverts xylulose and ribulose is D-tagatose 3-epimerase.

26. The cell of claim 25 , wherein the cell is an Escherichia coli cell.

Assignees

• Massachusetts Inst Technology

Inventors

• Stephanopoulos Gregory
• Pereira Brian
• De Mey Marjan
• Dugar Deepak
• Avalos Jose Luis

Classifications

• C12Y501/03

  acting on carbohydrates and derivatives (5.1.3) · CPC title

• C12Y401/02017

  L-Fuculose-phosphate aldolase (4.1.2.17) · CPC title

• C12Y207/01047

  D-Ribulokinase (2.7.1.47) · CPC title

• C12Y207/01016

  Ribulokinase (2.7.1.16) · CPC title

• C12Y102/01052

  Oxoglutarate dehydrogenase (NADP+) (1.2.1.52) · CPC title