Recombinant microorganisms capable of carbon fixation

Patent metadata

Field	Value
Publication number	US-10294482-B2
Application number	US-201515312734-A
Country	US
Kind code	B2
Filing date	May 21, 2015
Priority date	May 22, 2014
Publication date	May 21, 2019
Grant date	May 21, 2019

Abstract

A microorganism which is genetically modified so that it produces a first essential biomass precursor by metabolizing CO2 using a recombinant carbon fixation enzyme is disclosed. The microorganism produces a second biomass precursor by metabolizing an organic carbon source and not by metabolizing CO2. The microorganism does not use the organic carbon source for producing the first essential biomass precursor.

First claim

What is claimed is:

1. An E. coli which is genetically modified to express phosphoribulokinase (prk) and Ribulose-Bisphosphate Carboxylase (RuBisCo), wherein said E. coli produces a first essential biomass precursor by metabolizing CO 2 , wherein the E. coli produces a second biomass precursor by metabolizing an organic carbon source and not by metabolizing CO 2 , wherein the E. coli does not use said organic carbon source for producing said first essential biomass precursor, wherein said E. Coli is further modified so as to down-regulate expression or activity of phosphoglycerate mutase (gpm).

2. The microorganism of claim 1 , which generates ATP and NADH by metabolizing said organic carbon source.

3. The microorganism of claim 1 , wherein said first essential biomass precursor is derived solely from CO 2 .

4. The microorganism of claim 1 , wherein said first essential biomass precursor is derived from CO 2 and a second organic carbon source.

5. The microorganism of claim 1 , wherein said E. Coli is further modified so as to down-regulate expression or activity of phosphofructokinase (pfk) or ribose-5-phosphate isomerase (rpi).

6. The microorganism of claim 1 , wherein said E. Coli has a deletion in a gene selected from the group consisting of glucose 6-phosphate-1-dehydrogenase (zwf) 6-phosphogluconate dehydratase (edd) and ketohydroxyglutarate aldolase (eda), malate synthase A (aceB), isocitrate lyase (aceA) and isocitrate dehydrogenase kinase/phosphatase (aceK).

7. A cell culture comprising the microorganism of claim 1 and a medium comprising an organic carbon source.

8. A genetically modified E. Coli which expresses a recombinant phosphoribulokinase (prk) and Ribulose-Bisphosphate Carboxylase (RuBisCo) wherein the genes zwf is knocked-out or downregulated.

9. The genetically modified E. Coli of claim 8 , being an autotroph.

10. The genetically modified E. Coli of claim 8 , wherein the gene aceBAK and malate synthase G (glcB) are knocked-out or downregulated.

11. The genetically modified E. Coli of claim 8 , wherein the genes aceBAK, malate synthase G (glcB), phosphoenolpyruvate synthase (pps) and phosphoenolpyruvate carboxykinase (pck) are knocked-out or downregulated.

12. The genetically modified E. Coli of claim 8 , wherein the genes aceBAK and phosphopyruvate hydratase (eno) are knocked-out or downregulated.

13. The genetically modified E. Coli of claim 8 , wherein at least one gene selected from the group consisting of prs, yciV, topA, ydgC, slyA, dcm, malQ, brnQ, ppsR, truB, xylE and trmA is knocked-out or downregulated.

14. The genetically modified E. Coli of claim 8 , having at least one mutation as set forth in Tables 2-6.

15. The genetically modified E. Coli of claim 8 , wherein the gene pfkA, gpmA, gpmB and pfkB are knocked-out or down-regulated.

16. A cell culture comprising the genetically modified E. Coli of claim 8 and a medium comprising an organic carbon source.

Assignees

• Yeda Res & Dev

Inventors

• Milo Ron
• Antonovsky Niv
• Noor Elad
• Bar-Even Arren
• Zohar Yehudit
• Zelcbuch Lior
• Gleizer Shmuel
• Amram Shira

Classifications

• C12Y504/02

  Phosphotransferases (phosphomutases) (5.4.2) · CPC title

• C12N9/1025

  Acyltransferases (2.3) · CPC title

• C12N9/90

  Isomerases (5.) · CPC title

• C12Y101/01049

  Glucose-6-phosphate dehydrogenase (1.1.1.49) · CPC title

• C12Y207/01019

  Phosphoribulokinase (2.7.1.19) · CPC title