Microbial production of renewable glycolate

Patent metadata

Field	Value
Publication number	US-2017121717-A1
Application number	US-201615281994-A
Country	US
Kind code	A1
Filing date	Sep 30, 2016
Priority date	Oct 2, 2015
Publication date	May 4, 2017
Grant date	—

Abstract

Some aspects provide engineered microbes for glycolate production. Methods for microbe engineering and culturing are also provided herein. Such engineered microbes exhibit greatly enhanced capabilities for glycolate production.

First claim

What is claimed is:

1 . A cell engineered to produce glycolate, wherein the cell:
has reduced or eliminated activity of, or reduced or eliminated expression of xylulokinase relative to a wild type cell, recombinantly expresses an enzyme that interconverts xylulose and ribulose, recombinantly expresses a D-ribulose-phosphate aldolase, recombinantly expresses a D-ribulokinase, and recombinantly expresses a glycolaldehyde dehydrogenase.

2 .- 3 . (canceled)

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

5 .- 8 . (canceled)

9 . The cell of claim 1 , wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of glycolaldehyde reductase and/or glycolate oxidase or a subunit thereof relative to a wild type cell.

10 .- 12 . (canceled)

13 . The cell of claim 1 , wherein the cell further recombinantly expresses a glyoxylate reductase.

14 . (canceled)

15 . The cell of claim 1 , wherein the cell further recombinantly expresses isocitrate lyase.

16 . (canceled)

17 . The cell of claim 1 , wherein the cell further recombinantly expresses idh kinase/phosphatase, or wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of isocitrate lyase regulator relative to a wild type cell.

18 . (canceled)

19 . The cell of claim 1 , wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of glyoxylate carboligase and/or malate synthase A and/or malate synthase G relative to a wild type cell.

20 .- 24 . (canceled)

25 . The cell of claim 1 , wherein the cell is a bacterial cell.

26 .- 29 . (canceled)

30 . An Escherichia coli cell engineered to produce glycolate, wherein the cell comprises a deletion of a xylB gene, recombinantly expresses a dte gene from Pseudomonas cichorii that is codon-optimized for E. coli , recombinantly expresses a fucA gene from E. coli , recombinantly expresses a fucK gene from E. coli , and recombinantly expresses an aldA gene from E. coli.

31 . The Escherichia coli of claim 30 , wherein the cell further comprises a deletion of a fucO gene.

32 . The Escherichia coli cell of claim 30 , wherein the cell further comprises a deletion of a glcD gene.

33 . The Escherichia coli cell of claim 30 , wherein the cell further recombinantly expresses a ycdW gene from E. coli.

34 . The Escherichia coli cell of claim 30 , wherein the cell further recombinantly expresses an aceA gene from E. coli.

35 . The Escherichia coli cell of claim 30 , wherein the cell further recombinantly expresses an aceK gene from E. coli , or wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of isocitrate lyase regulator relative to a wild type cell.

36 . The Escherichia coli cell of claim 30 , wherein the cell further comprises a deletion of a gcl gene and/or a aceB gene and/or a glcB gene.

37 .- 45 . (canceled)

46 . The cell of claim 19 , wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of malate synthase A relative to a wild type cell.

47 . The cell of claim 46 , wherein the cell has reduced or eliminated activity of, or reduced or eliminated expression of glyoxylate carboligase relative to a wild type cell.

48 . The Escherichia coli cell of claim 32 , wherein the cell further comprises a deletion of a fucO gene.

49 . The Escherichia coli cell of claim 36 , wherein the cell further comprises a deletion of an aceB gene.

50 . The Escherichia coli cell of claim 49 , wherein the cell further comprises a deletion of a gcl gene.

Assignees

• Massachusetts Inst Technology

Inventors

• Stephanopoulos Gregory
• Li zheng-jun
• Pereira Brian

Classifications

• C12N9/0008

  acting on the aldehyde or oxo group of donors (1.2) · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12Y102/01021

  Glycolaldehyde dehydrogenase (1.2.1.21) · CPC title

• C12N9/90

  Isomerases (5.) · CPC title

• C12Y401/03001

  Isocitrate lyase (4.1.3.1) · CPC title