Pyruvate carboxylase and pyruvate carboxylase-encoding DNA, plasmid containing said DNA and microorganism for the production thereof, and methods for the production of products the biosynthesis of which includes oxaloacetate as precursor, and chromosome

Patent metadata

Field	Value
Publication number	US-11028384-B2
Application number	US-201816606230-A
Country	US
Kind code	B2
Filing date	Apr 13, 2018
Priority date	May 18, 2017
Publication date	Jun 8, 2021
Grant date	Jun 8, 2021

Abstract

A DNA sequence that includes at least 70% identity with respect to SEQ ID NO: 1 and further includes a triplet at position 1027-1029 that codes for alanine.

First claim

The invention claimed is:

1. An isolated DNA sequence encoding a pyruvate carboxylase, comprising:
at least 97% identity with respect to SEQ ID NO: 1,
wherein a triplet at position 1027-1029 codes for alanine.

2. The DNA sequence according to claim 1 , wherein the DNA has identity of at least 99% with respect to SEQ ID NO: 1.

3. The DNA sequence according to claim 1 , wherein the DNA sequence is a DNA according to SEQ ID NO: 1.

4. The DNA sequence according to claim 1 , further comprising, at position 3034-3036, a triplet which encodes for serine.

5. The DNA sequence according to claim 4 , wherein the DNA has identity of at least 97% with respect to SEQ ID NO: 2.

6. The DNA sequence according to claim 4 , wherein the DNA is according to SEQ ID NO: 2.

7. A pyruvate carboxylase having at least 97% identity with respect to a pyruvate carboxylase according to SEQ ID NO: 3, wherein alanine is present at position 343.

8. The pyruvate carboxylase according to claim 7 , wherein the pyruvate carboxylase has identity of at least 99% with respect to the pyruvate carboxylase according to SEQ ID NO: 3.

9. The pyruvate carboxylase according to claim 7 , wherein the pyruvate carboxylase is a pyruvate carboxylase according to SEQ ID NO: 3.

10. The pyruvate carboxylase according to claim 7 , further comprising serine at position 1012.

11. The pyruvate carboxylase according to claim 10 , wherein the pyruvate carboxylase has identity of at least 97% with respect to the pyruvate carboxylase according to SEQ ID NO: 4.

12. The pyruvate carboxylase according to claim 10 , wherein the pyruvate carboxylase is a pyruvate carboxylase according to SEQ ID NO: 4.

13. A vector, wherein the vector comprises a DNA sequence according to claim 1 .

14. The vector according to claim 13 , wherein the vector is a plasmid.

15. A microorganism, wherein the microorganism comprises a DNA sequence according to claim 1 .

16. The microorganism according to claim 15 , wherein the microorganism comprises a vector.

17. The microorganism according to claim 15 , wherein the microorganism is a microorganism selected from the group consisting of the genera Corynebacterium, Brevibacterium, Bacillus, Lactobacillus, Lactococcus, Candida, Pichia, Kluyveromyces, Saccharomyces, Escherichia, Zymomonas, Yarrowia, Methylobacterium, Ralstonia, Vibrio , and Clostridium.

18. A method for the production of products whose biosynthesis includes oxaloacetate as a precursor, the method comprising:
fermenting a microorganism containing a DNA sequence according to claim 1 , which forms a pyruvate carboxylase, and
performing an enrichment of the products in a medium or in cells.

19. The method according to claim 18 , wherein a product is isolated.

20. The method according to claim 18 , wherein a species from the group consisting of the genera Corynebacterium, Brevibacterium, Bacillus, Lactobacillus, Lactococcus, Candida, Pichia, Kluyveromyces, Saccharomyces, Escherichia, Zymomonas, Yarrowia, Methylobacterium, Ralstonia, Vibrio , and Clostridium is used as the microorganism.

21. The method according to claim 18 , wherein an amino acid of the aspartate family is produced.

22. The method according to claim 21 , wherein L-lysine, L-aspartate, L-asparagine, L-threonine, L-isoleucine, or L-methionine is produced.

23. The method according to claim 18 , wherein substances from the glutamate family of amino acids are produced, such as L-glutamate, glutamine, L-arginine, L-proline, L-citrulline, or L-ornithine; intermediates of the citrate cycle, such as malate, fumarate, succinate, malate, fumarate citrate, isocitrate, or 2-oxoglutarate; diamines, such as diaminopentane or diaminobutane; as well as further metabolites prepared from oxaloacetate, such itaconate, ectoine, gamma aminobutyrate, butanol, 1-propanol, L-citrulline, L-ornithine, D-arginine, or 4-hydroxyproline.

24. The method according to claim 18 , wherein the DNA sequence is more strongly expressed.

25. A chromosome including the DNA sequence according to claim 1 .

Assignees

• Forschungszentrum Juelich Gmbh

Inventors

• Kortmann Maike
• Baumgart Meike
• Bott Michael

Classifications

• C12N1/205

  Bacterial isolates · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12Y604/01001

  Pyruvate carboxylase (6.4.1.1) · CPC title

• C12N9/93 Primary

  Ligases (6) · CPC title

• C12N9/1217

  Phosphotransferases with a carboxyl group as acceptor (2.7.2) · CPC title