Process for producing an organic compound

Patent metadata

Field	Value
Publication number	US-11279955-B2
Application number	US-201816614772-A
Country	US
Kind code	B2
Filing date	May 18, 2018
Priority date	May 19, 2017
Publication date	Mar 22, 2022
Grant date	Mar 22, 2022

Abstract

Described herein is a process of producing an organic compound, the process including: I) cultivating a genetically modified microorganism in a culture medium including sucrose as an assimilable carbon source to allow the genetically modified microorganism to produce the organic compound, and II) recovering the organic compound from the fermentation broth obtained in process step I) The genetically modified microorganism includes A) at least one genetic modification that leads to an increased activity of the enzyme encoded by the rbsK-gene, compared to the original microorganism that has not been genetically modified, and the original microorganism belongs to the family Pasteurellaceae. Also described herein are a genetically modified microorganism and the use thereof for the fermentative production of an organic compound from sucrose as an assimilable carbon source.

First claim

The invention claimed is:

1. A process of producing an organic compound, the process comprising:
I) cultivating a genetically modified microorganism in a culture medium comprising sucrose as an assimilable carbon source to allow the genetically modified microorganism to produce the organic compound, and
II) recovering the organic compound from the fermentation broth obtained in process step I),
wherein the genetically modified microorganism comprises:
C) at least one genetic modification that leads to an increased activity of the enzyme encoded by the rbsK-gene, compared to the original microorganism that has not been genetically modified,
and wherein the original microorganism belongs to the family Pasteurellaceae.

2. The process according to claim 1 , wherein the organic compound is succinic acid.

3. The process according to claim 1 , wherein the original microorganism belongs to the genus Basfia.

4. The process according to claim 3 , wherein the original microorganism belongs to the species Basfia succiniciproducens.

5. The process according to claim 4 , wherein the original microorganism is Basfia succiniciproducens strain DD1 as deposited under DSM 18541 with the DSMZ, Germany.

6. The process according to claim 1 , wherein the rbsK-gene comprises a nucleic acid selected from the group consisting of:
a1) nucleic acids having the nucleotide sequence of SEQ ID NO: 3;
b1) nucleic acids encoding the amino acid sequence of SEQ ID NO: 4;
c1) nucleic acids which are at least 70% identical to the nucleic acid of a1) or b1), the identity being the identity over the total length of the nucleic acids of a1) or b1);
d1) nucleic acids encoding an amino acid sequence which is at least 70% identical to the amino acid sequence encoded by the nucleic acid of a1) or b1), the identity being the identity over the total length of amino acid sequence encoded by the nucleic acids of a1) or b1);
e1) nucleic acids capable of hybridizing under stringent conditions with a complementary sequence of any of the nucleic acids according to a1) or b1), wherein the stringent conditions are conditions wherein 100 contiguous nucleotides or more, which are a fragment or identical to the complementary sequence of any of the nucleic acids according to a1) or b1), hybridize under conditions equivalent to hybridization in 7% SDS, 0.5 M NaPO4, 1 mM EDTA at 50° C., with washing in 2×SSC, 0.1% SDS at 50° C.; and
f1) nucleic acids encoding the same protein as any of the nucleic acids of a1) or b1), but differing from the nucleic acids of a1) or b1) above due to the degeneracy of the genetic code.

7. The process according to claim 1 , wherein the at least one genetic modification C) comprises a modification of the rbsK-gene, a modification of a regulatory element of the rbsK-gene, or a combination of both.

8. The process according to claim 7 , wherein the at least one genetic modification C) comprises an overexpression modification of the rbsK-gene.

9. The process according to claim 1 , wherein the genetically modified microorganism additionally comprises:
D) at least one genetic modification that leads to a reduced activity of the enzyme encoded by the fruA-gene, compared to the original microorganism that has not been genetically modified.

10. The process according to claim 9 , wherein the fruA-gene comprises a nucleic acid selected from the group consisting of:
a2) nucleic acids having the nucleotide sequence of SEQ ID NO: 5;
b2) nucleic acids encoding the amino acid sequence of SEQ ID NO: 6;
c2) nucleic acids which are at least 70% identical to the nucleic acid of a2) or b2), the identity being the identity over the total length of the nucleic acids of a2) or b2);
d2) nucleic acids encoding an amino acid sequence which is at least 70% identical to the amino acid sequence encoded by the nucleic acid of a2) or b2), the identity being the identity over the total length of amino acid sequence encoded by the nucleic acids of a2) or b2);
e2) nucleic acids capable of hybridizing under stringent conditions with a complementary sequence of any of the nucleic acids according to a2) or b2), wherein the stringent conditions are conditions wherein 100 contiguous nucleotides or more, which are a fragment or identical to the complementary sequence of any of the nucleic acids according to a2) or b2), hybridize under conditions equivalent to hybridization in 7% SDS, 0.5 M NaPO4, 1 mM EDTA at 50° C., with washing in 2×SSC, 0.1% SDS at 50° C.; and
f2) nucleic acids encoding the same protein as any of the nucleic acids of a2) or b2), but differing from the nucleic acids of a2) or b2) above due to the degeneracy of the genetic code.

11. The process according to claim 10 , wherein the at least one genetic modification D) comprises a modification of the fruA-gene, a modification of a regulatory element of the fruA-gene, or a combination of both.

12. The process according to claim 11 , wherein the at least one modification D) comprises an inactivation of the fruA-gene.

13. A genetically modified microorganism that comprises:
A) at least one genetic modification that leads to an increased activity of the enzyme encoded by the rbsK-gene, compared to the original microorganism that has not been genetically modified,
wherein the original microorganism belongs to the family Pasteurellaceae.

14. The genetically modified microorganism according to claim 13 , wherein the at least one genetic modification A) comprises a modification of the rbsK-gene, a modification of a regulatory element of the rbsK-gene, or a combination of both.

15. The genetically modified microorganism according to claim 14 , wherein the at least one genetic modification A) comprises an overexpression modification of the rbsK-gene.

16. The genetically modified microorganism according to claim 13 , further comprising:
B) at least one genetic modification that leads to a reduced activity of the enzyme encoded by the fruA-gene, compared to the original microorganism that has not been genetically modified.

17. The genetically modified microorganism according to claim 16 , wherein the fruA-gene comprises a nucleic acid selected from the group consisting of:
a2) nucleic acids having the nucleotide sequence of SEQ ID NO: 5;
b2) nucleic acids encoding the amino acid sequence of SEQ ID NO: 6;
c2) nucleic acids which are at least 70% identical to the nucleic acid of a2) or b2), the identity being the identity over the total length of the nucleic acids of a2) or b2);
d2) nucleic acids encoding an amino acid sequence which is at least 70% identical to the amino acid sequence encoded by the nucleic acid of a2) or b2), the identity being the identity over the total length of amino acid sequence encoded by the nucleic acids of a2) or b2);
e2) nucleic acids capable of hybridizing under stringent conditions with a complementary sequence of any of the nucleic acids according to a2) or b2), wherein the stringent conditions are conditions wherein 100 contiguous nucleotides or more, which are a fragment or identical to the complementary sequence of any of the nucleic acids according to a2) or b2), hybridize under conditions equivalent to hybridization in 7% SDS, 0.5 M NaPO4, 1 mM EDTA at 50° C., with washing in 2×SSC, 0.1% SDS at 50° C.; and
f2) nucleic acids encoding the same protein as any of the nucleic acids of a2) or b2), but differing from the nucleic acids of a2) or b2) above due to the degeneracy of the genetic code.

18. The genetically modified microorganism according to claim 17 , wherein the at least one genetic modification B)

Assignees

• Basf Se

Inventors

• Schroeder Hartwig
• Haefner Stefan
• Zelder Oskar
• Wittmann Christoph
• Schroer Anna Christine
• Hoff Birgit
• Boehmer Nico

Classifications

• C12P7/46 Primary

  Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid · CPC title

• C12R2001/01

  Bacteria or Actinomycetales {; using bacteria or Actinomycetales} · CPC title

• C12Y207/01005

  Rhamnulokinase (2.7.1.5) · CPC title

• C12N9/1205 Primary

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title

• C12P7/18 Primary

  polyhydric · CPC title