Serinol production in glycerol catabolism deficient Escherichia coli strains

Patent metadata

Field	Value
Publication number	US-9109240-B2
Application number	US-201314018725-A
Country	US
Kind code	B2
Filing date	Sep 5, 2013
Priority date	Sep 14, 2012
Publication date	Aug 18, 2015
Grant date	Aug 18, 2015

Abstract

The invention is about E. coli host cells which are capable to convert glycerol to serinol. Furthermore, a process for producing serinol is disclosed, which comprises culturing E. coli host cells inactive for triosephosphate isomerase and active for dihydroxyacetone phosphate aminotransferase to convert glycerol to serinol, induction of conversion from glycerol to serinol by adding at least glycerol to the cell culture, and isolating serinol from the cell culture.

First claim

The invention claimed is:

1. A recombinant E. coli strain active for dihydroxyacetone phosphate aminotransferase capable to convert glycerol to serinol, wherein triosephosphate isomerase is inactivated in the E. coli host cells.

2. The recombinant E. coli strain according to claim 1 , wherein methylglyoxal synthase is inactivated additionally in the E. coli host cells.

3. The recombinant E. coli strain according to claim 1 , wherein the glp DNA-binding transcriptional repressor is inactivated additionally in the E. coli host cells.

4. The recombinant E. coli strain according to claim 1 , wherein active dihydroxyacetone phosphate aminotransferase is introduced into the said strain by transforming said strain with an expression vector comprising a transgene coding for dihydroxyacetone phosphate aminotransferase which is active for conversion of glycerol to serinol and wherein the expression of the dihydroxyacetone phosphate aminotransferase is inducible.

5. A recombinant E. coli strain selected from the group consisting of
a. Strain E. coli MG1655ΔtpiA,
b. Strain E. coli MG1655ΔtpiAΔmgsA,
c. Strain E. coli MG1655ΔtpiAΔglpR, and
d. Strain E. coli MG1655ΔtpiAΔmgsA ΔglpR,
wherein said strain further comprises a gene encoding dihydroxyacetone phosphate aminotransferase(rtxA) and said rtxA is regulated by a natural promoter of tpiA and wherein said dihydroxyacetone phosphate aminotransferase is active for conversion of glycerol to serinol.

6. A recombinant E. coli strain selected from the group consisting of
a. Strain E. coli HMS174(DE3)ΔtpiA,
b. Strain E. coli HMS174(DE3)ΔtpiAΔmgsA,
c. Strain E. coli HMS174(DE3)ΔtpiAΔglpR,
d. Strain E. coli HMS174(DE3)ΔtpiAΔmgsAΔglpR, and

wherein the strains are transformed with pCOLADuet-1::rtxA and wherein rtxA codes for dihydroxyacetone phosphate aminotransferase which is active for conversion of glycerol to serinol.

7. The recombinant E. coli strain according to claim 6 wherein the strains are transformed with pCOLADuet-1::rtxD::rtxA.

8. The recombinant E. coli strain according to claim 6 wherein the strains are transformed with pCOLADuet-1::rtxEFG::rtxA.

9. The recombinant E. coli strain according to claim 6 wherein the strains are transformed with pCOLADuet-1::rtxDEFG::rtxA.

10. A process for producing serinol, which comprises
i) culturing E. coli host cells inactive for triosephosphate isomerase and active for dihydroxyacetone phosphate aminotransferase to convert glycerol to serinol,
ii) induction of conversion from glycerol to serinol by adding at least glycerol to the cell culture, and
iii) isolating serinol from the cell culture.

11. The process according to claim 10 , wherein the E. coli host cells are in addition inactive for methylglyoxal synthase.

12. The process according to claim 10 , wherein glp DNA-binding transcriptional repressor is inactivated additionally in the E. coli host cells.

13. The process according to claim 10 , wherein expression of active dihydroxyacetone phosphate aminotransferase is achieved by introducing an expression vector into the host cells comprising the transgene coding for dihydroxyacetone phosphate aminotransferase which is active for conversion of glycerol to serinol.

Assignees

• Basf Se
• Univ Muenster Wilhelms
• Univ Muenster Wilhelms

Inventors

• Maksym Lukas
• Steinbuechel Alexander
• Andreessen Bjoern

Classifications

• C12N9/1096

  transferring nitrogenous groups (2.6) · CPC title

• C12P13/001 Primary

  Amines; Imines · CPC title

• C12N9/90

  Isomerases (5.) · CPC title