IP Library Granted Patent US 9,994,876
Granted Patent B2
US 9,994,876 · App. 14/864,493 · Granted Jun 12, 2018

Engineering microbes and metabolic pathways for the production of ethylene glycol

Inventors: Gregory Stephanopoulos (Winchester, MA); Brian Pereira (Cambridge, MA); Marjan De Mey (Ghent, BE); Deepak Dugar (Cambridge, MA); Jose Luis Avalos (Arlington, MA)
Assignee: Massachusetts Institute of Technology
C12P7/24C12N9/0006C12N9/0008C12N9/0022C12N9/1096C12N9/1205C12N9/1229C12N9/16C12N9/88C12N9/90C12N15/52C12N15/70C12P7/18C12Y101/01071C12Y102/01052C12Y207/01016C12Y207/01047C12Y401/02017C12Y501/03C12P2203/00C12Y102/01021C12Y207/01017
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Quick Facts
Patent No.
US 9,994,876
App. No.
14/864,493
Granted
Jun 12, 2018
Kind
B2
Abstract

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

Claims (26)

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.

Assignments (2)
CONFIRMATORY LICENSE Recorded Dec 15, 2023
From: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
To: U.S. DEPARTMENT OF ENERGY
Reel/Frame 066045/0525 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 5, 2016
From: STEPHANOPOULOS, GREGORY; PEREIRA, BRIAN; DE MEY, MARJAN; DUGAR, DEEPAK; AVALOS, JOSE LUIS
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 037408/0715 →
Continuity (3)
Continuation 13774212 · Feb 22, 2013
Provisional Application 61602322 · Feb 23, 2012
Related Publication 20160076061A1 · Mar 17, 2016