Process for preparing L amino acids using improved strains of the enterobacteriaceae family

Patent metadata

Field	Value
Publication number	US-11053526-B2
Application number	US-201916533841-A
Country	US
Kind code	B2
Filing date	Aug 7, 2019
Priority date	Aug 9, 2018
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

The invention relates to a recombinant, L-amino acid-secreting microorganism of the Enterobacteriaceae family, comprising an DNA fragment having promoter activity that is functionally linked to a polynucleotide coding for a membrane protein, characterized in that the DNA fragment having promoter activity comprises the SEQ ID NO: 8.

First claim

What is claimed is:

1. A recombinant L-amino acid-secreting microorganism of the Enterobacteriaceae family, comprising an DNA fragment having promoter activity that is functionally linked to a polynucleotide coding for a membrane protein, wherein the DNA fragment having promoter activity comprises the sequence of SEQ ID NO: 8.

2. The microorganism of claim 1 , wherein the DNA fragment having promoter activity is linked at the 3′ end to a second DNA fragment carrying a ribosome-binding site.

3. The microorganism of claim 1 , wherein the DNA fragment having promoter activity is linked at the 3′ end to a second DNA fragment comprising the nucleotide sequence of positions 174 to 204 of SEQ ID NO: 9.

4. The microorganism of claim 1 , wherein the DNA fragment having promoter activity is linked at the 3′ end to a second DNA fragment having the nucleotide sequence of positions 174 to 204 of SEQ ID NO: 9 which is linked at its 3′ end to a polynucleotide coding for the membrane protein.

5. The microorganism of claim 1 , wherein the DNA fragment having promoter activity is linked at the 5′ end to a DNA fragment having the nucleotide sequence of positions 1 to 138 of SEQ ID NO: 9.

6. The microorganism of claim 1 , wherein the membrane protein has the activity of an amino acid transporter.

7. The microorganism of claim 6 , wherein the protein having the activity of an amino acid transporter is a protein having the activity of an amino acid exporter.

8. The microorganism of claim 7 , wherein the protein having the activity of an amino acid exporter has an amino acid sequence which is at least 90% identical to the sequence of SEQ ID NO: 2.

9. The microorganism of claim 8 , wherein the protein having the activity of an amino acid exporter has an amino acid sequence, which is at least 95% identical to the sequence of SEQ ID NO: 2.

10. The microorganism of claim 9 , wherein the protein having the activity of an amino acid exporter comprises the amino acid sequence of SEQ ID NO: 2, and/or is encoded by a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1.

11. The microorganism of claim 1 , wherein the DNA fragment having promoter activity is present in the chromosome of the microorganism, or, alternatively, the DNA fragment having promoter activity is located on an extrachromosomal replicating vector.

12. The microorganism of claim 1 , wherein said microorganism produces L-threonine, L-homoserine, L histidine, L-lysine, L-tryptophan, L valine, L-leucine, and L-isoleucine.

13. The microorganism of claim 4 , wherein the DNA fragment having promoter activity is linked at the 5′ end to a DNA fragment having the nucleotide sequence of positions 1 to 138 of SEQ ID NO: 9.

14. The microorganism of claim 13 , wherein the membrane protein has the activity of an amino acid exporter and comprises the amino acid sequence of SEQ ID NO: 2, and/or is encoded by a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1.

15. A process for preparing L amino acids or feedstuff additives containing L-amino acids, the process comprising:
(i) fermenting the microorganism of claim 1 in a medium;
(ii) enriching the L-amino acid in the fermentation medium and/or in the cell; and optionally
(iii) isolating the L-amino acid.

16. A DNA fragment having promoter activity that is functionally linked to a polynucleotide coding for a membrane protein, wherein the DNA fragment having promoter activity comprises SEQ ID NO: 8.

Assignees

• Evonik Operations Gmbh

Inventors

• Rieping Mechthild

Classifications

• A23K10/12 Primary

  by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass · CPC title

• C12P13/08

  Lysine; Diaminopimelic acid; Threonine; Valine · CPC title

• C12P13/06

  Alanine; Leucine; Isoleucine; Serine; Homoserine · CPC title

• A23V2002/00

  Food compositions, function of food ingredients or processes for food or foodstuffs · CPC title

• A23K20/142

  Amino acids; Derivatives thereof · CPC title