Microorganism and method for production of amino acids by fermentation

Patent metadata

Field	Value
Publication number	US-2016265072-A2
Application number	US-201414762840-A
Country	US
Kind code	A2
Filing date	Jan 13, 2014
Priority date	Jan 30, 2013
Publication date	Sep 15, 2016
Grant date	—

Abstract

The invention relates to a microorganism that produces and/or secretes an organic chemical compound, wherein the microorganism has an increased expression, compared to the respective starting strain, of a polypeptide LpdA with the activity of a transhydrogenase; and a process for producing an organic chemical compound using the microorganism according to the invention.

First claim

1 . Process for producing an organic chemical compound by fermentation using a microorganism, characterized in that the following steps are carried out:
a) fermentation of a microorganism producing an organic chemical compound, wherein a polynucleotide that codes for a polypeptide whose amino acid sequence is at least 80% identical to the amino acid sequence of SEQ ID NO:2 is overexpressed in the microorganism and wherein the fermentation takes place in a fermentation medium, with formation of a fermentation broth, b) enrichment of the organic chemical compound in the fermentation broth from a).

2 . Process for producing an organic chemical compound according to claim 1 , characterized in that a polynucleotide that codes for a polypeptide whose amino acid sequence is at least 95% identical to the amino acid sequence of SEQ ID NO:2 is overexpressed in the microorganism.

3 . Process for producing an organic chemical compound according to claim 1 , characterized in that the encoded polypeptide has the activity of a transhydrogenase.

4 . Process for producing an organic chemical compound according to claim 1 , characterized in that the encoded polypeptide comprises the amino acid sequence of SEQ ID NO:2.

5 . Process for producing an organic chemical compound according to claim 1 , characterized in that the production of the organic chemical compound is increased by at least 0.5% relative to the fermentation of a microorganism in which the polynucleotide that codes for a polypeptide whose amino acid sequence is at least 80% identical to the amino acid sequence of SEQ ID NO:2 is not overexpressed.

6 . Process for producing an organic chemical compound according to claim 1 , characterized in that the overexpression is achieved by one or more of the measures selected from the group
a) expression of the gene is under the control of a promoter, which is stronger in the microorganism used for the process than the native promoter of the gene; b) increase of the copy number of the gene coding for a polypeptide with the activity of a transhydrogenase; preferably by inserting the gene in plasmids with increased copy number and/or by integrating the gene into the chromosome of the microorganism in at least one copy; c) expression of the gene takes place using a ribosome binding site, which is stronger in the microorganism used for the process than the original ribosome binding site of the gene; d) expression of the gene takes place with optimization of the codon usage of the microorganism used for the process; e) expression of the gene takes place with reduction of mRNA secondary structures in the mRNA transcribed by the gene; f) expression of the gene takes place with elimination of RNA polymerase terminators in the mRNA transcribed by the gene; g) expression of the gene takes place using mRNA-stabilizing sequences in the mRNA transcribed by the gene.

7 . Process for producing an organic chemical compound according to claim 1 , characterized in that the organic chemical compound is an L-amino acid selected from the group L-threonine, L-isoleucine, L-lysine, L-methionine, L-ornithine, L-proline, L-valine, L-leucine and L-tryptophan.

8 . Process for producing an organic chemical compound according to claim 1 , characterized in that it is an organic chemical compound-secreting microorganism of the genus Corynebacterium or Escherichia.

9 . Process for producing an organic chemical compound according to claim 1 , characterized in that it is an organic chemical compound-secreting microorganism of the species Corynebacterium glutamicum or of the species Escherichia coli.

10 . Process for producing an organic chemical compound according to claim 1 , characterized in that it is a process selected from the group: batch process, fed-batch process, repeated fed-batch process and continuous process.

11 . Process according to claim 7 , characterized in that the organic chemical compound or a liquid or solid organic chemical compound-containing product is obtained from the organic chemical compound-containing fermentation broth.

12 . Microorganism that produces an organic chemical compound, in which a polynucleotide that codes for a polypeptide whose amino acid sequence is ≧80% identical to the amino acid sequence of SEQ ID NO: 2 is overexpressed.

13 . Microorganism according to claim 12 , characterized in that the amino acid sequence of the encoded polypeptide is ≧95% identical to the amino acid sequence of SEQ ID NO: 2.

14 . Microorganism according to claim 12 , characterized in that the encoded polypeptide comprises the amino acid sequence of SEQ ID NO: 2.

15 . Microorganism according to claim 12 , selected from the group of the genus Corynebacterium and the bacteria of the family Enterobacteriaceae, wherein the species Corynebacterium glutamicum is preferred.

16 . Microorganism according to claim 12 , characterized in that the overexpressed polynucleotide has the activity of a transhydrogenase.

Assignees

• Evonik Degussa Gmbh

Inventors

• Walter Frederik
• Persicke Marcus
• Kalinowski Jorn
• Huser Andrea
• Claes Wilfried
• Reth Alexander

Classifications

• C12R1/15 Primary

  Chemistry & Metallurgy · mapped topic

• C12P13/12

  Methionine; Cysteine; Cystine · CPC title

• C12P13/08

  Lysine; Diaminopimelic acid; Threonine; Valine · CPC title

• C12N9/0051 Primary

  acting on a sulfur group of donors (1.8) · CPC title

• C12N1/205 Primary

  Bacterial isolates · CPC title