Process for the fermentative preparation of sulphur-containing amino acids

Patent metadata

Field	Value
Publication number	US-9562245-B2
Application number	US-201514712596-A
Country	US
Kind code	B2
Filing date	May 14, 2015
Priority date	Jan 20, 2011
Publication date	Feb 7, 2017
Grant date	Feb 7, 2017

Abstract

The invention relates to a process for the fermentative preparation of sulphur-containing amino acids chosen from the group of L-methionine, L-cysteine, L-cystine, L-homocysteine and L-homocystine, comprising the steps:
a) provision of a microorganism of the family Enterobacteriaceae or of a microorganism of the family Corynebacteriaceae which has an increased thiosulphate sulphurtransferase activity compared with the particular starting strain; b) fermentation of the microorganism, from a) in a medium which contains an inorganic source of sulphur chosen from the group of salt of dithiosulphuric acid or a mixture of a salt, of dithiosulphuric acid and a salt of sulphuric acid, a fermentation broth being obtained, and c) concentration of the sulphur-containing amino acid in the fermentation broth from b).

First claim

The invention claimed is:

1. A process for the fermentative preparation of sulphur-containing amino acids selected from the group consisting of L-methionine, L-cysteine, L-cystine, L-homocysteine and L-homocystine, comprising the steps:
a) provision of a microorganism of the family Enterobacteriaceae or of a microorganism of the family Corynebacteriaceae which overexpresses a gene coding for a polypeptide with the activity of a thiosulphate sulphurtransferase comprising the amino acid sequence shown in SEQ ID NO: 2 or an amino acid sequence containing a deletion, substitution, insertion and/or addition of from 1 to 45 amino acid residues with respect to the amino acid sequence shown in SEQ ID NO: 2;
b) fermentation of the microorganism from a) in a medium which contains an inorganic source of sulphur from the group of salt of thiosulphuric acid or a mixture of a salt of thiosulphuric acid and a salt of sulphuric acid, a fermentation broth being obtained, and
c) concentration of the sulphur-containing amino acid in the fermentation broth from b).

2. The process according to claim 1 , wherein the expression of the gene coding for a polypeptide with the activity of a thiosulphate sulphurtransferase is increased by one or more measures selected from the group consisting of:
a) the expression of the gene is under the control of a promoter which, in the microorganism used for the process, leads to an amplified expression compared with the starting strain;
b) increasing the number of copies of the gene coding for a polypeptide with the activity of a thiosulphate sulphurtransferase compared with the starting strain by insertion of the gene into plasmids or by integration of the gene into the chromosome of the microorganism in several copies;
c) the expression of the gene is effected using a ribosome binding site, which leads to an increased translation in the microorganism used for the process compared with the starting strain;
d) the expression of the gene is amplified by an optimization of the codon usage with respect to the microorganism used for the process;
e) the expression of the gene is amplified by reduction of mRNA secondary structures in the mRNA transcribed by the gene;
f) the expression of the gene is amplified by elimination of RNA polymerase terminators in the mRNA transcribed by the gene;
g) the expression of the gene is effected using mRNA-stabilizing sequences in the mRNA transcribed by the gene.

3. The process according to claim 1 , wherein the salt of thiosulphuric acid is a salt chosen from the group of alkali metal salt, alkaline earth metal salt, ammonium salt and mixtures thereof.

4. The process according to claim 1 , wherein the salt of sulphuric acid is a salt selected from the group consisting of alkali metal salt, alkaline earth metal salt, ammonium salt and mixtures thereof.

5. The process according to claim 1 , wherein during the fermentation the content of the salt of thiosulphuric acid, based on the total content of inorganic sulphur in the medium and in the fermentation broth, is kept at at least 5 mol %.

6. The process according to claim 1 , wherein the sulphur-containing amino acid is L-methionine.

7. The process according to claim 1 , wherein the microorganism is of the genus Escherichia.

8. The process according to claim 7 , wherein the species of the microorganism of the genus Escherichia is Escherichia coli.

9. The process according to claim 1 , wherein the microorganism is the genus Corynebacterium.

10. The process according to claim 9 , wherein the species of the microorganism is Corynebacterium glutamicum.

11. The process according to claim 1 , wherein the microorganism is selected from the group consisting of
Corynebacterium glutamicum with increased activity and/or expression of aspartate kinase and attenuation or deletion of the regulator protein McbR compared with the starting strain; and
Escherichia coli with increased activity and/or expression of aspartate kinase and attenuation or deletion of the regulator protein MetJ compared with the starting strain.

Assignees

• Evonik Degussa Gmbh

Inventors

• Schneider Frank
• Molck Stella
• Bathe Brigitte

Classifications

• C12Y208/01001

  Thiosulfate sulfurtransferase (2.8.1.1) · CPC title

• C12P13/12 Primary

  Methionine; Cysteine; Cystine · CPC title

• C12N9/13

  transferring sulfur containing groups (2.8) · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title

• C12R1/15

  Chemistry & Metallurgy · mapped topic