Processes for the fermentative preparation of L-threonine using strains of
The invention relates to a process for the preparation of L-amino acids, in particular L-threonine, in which the following steps are carried out: a) fermentation of microorganisms of the Enterobacteriaceae family which produce the desired L-amino acid and in which the yjgF ORF or the nucleotide sequence which codes for it are attenuated, in particular eliminated, b) concentration of the L-amino acid in the medium or in the cells of the bacteria, and optionally c) isolation of the L-amino acid.
1. A process for producing L-threonine comprising:
a) culturing or fermenting an Escherichia coli cell in a medium for a time and under conditions suitable for producing said L-threonine;
b) isolating said L-threonine; and
c) determining the amount of L-threonine isolated,
wherein the expression of the yjgF open reading frame of said E. coli cell is eliminated by one or more methods of mutagenesis selected from the group consisting of: deletion of all or at least one nucleotide of the yjgF open reading frame; insertion of at least one nucleotide in the yjgF open reading frame; and transitional or transversional mutagenesis with incorporation of a non-sense mutation in the yjgF open reading frame, wherein said elimination of the expression of the yjgF open reading frame results in an increased production of L-threonine by said E. coli cell relative to the amount of L- threonine produced in said E. coli cell prior to said elimination of the expression of the yjgF open reading frame; and wherein prior to elimination of the expression of the yjgF open reading frame, said yjgF open reading frame comprises the nucleotide sequence of SEQ ID NO: 1 and encodes the polypeptide of SEQ ID NO:2.
2. The process of claim 1 , wherein the expression of the yjgF open reading frame has been eliminated by the deletion of at least one nucleotide of the yjgF open reading frame.
3. The process of claim 1 , wherein said L-threonine is isolated from said E. coli.
4. The process of claim 1 , wherein said L-threonine is isolated from said medium.
5. The process of claim 1 , wherein culturing or fermenting is performed using a batch process.
6. The process of claim 1 , wherein culturing or fermenting is performed using a fed batch process.
7. The process of claim 1 , wherein culturing or fermenting is performed using a repeated fed batch process.
8. A process for producing L-threonine, comprising:
a) culturing or fermenting an Escherichia coli cell in a medium for a time and under conditions suitable for producing said L-threonine; and
b) recovering said L-threonine and determining the amount of said L-threonine recovered,
wherein the expression of the yjgF open reading frame of said E. coli has been eliminated by deletion of all of the yjgF open reading frame and wherein said yjgF open reading frame encodes the polypeptide of SEQ ID NO:2.
9. The process of claim 8 , wherein said yjgF open reading frame has the nucleotide sequence of SEQ ID NO:1.
10. The process of claim 8 , wherein constituents of the culture or fermentation broth and/or biomass in its entirety or portions thereof remain with the recovered L-threonine of step b).
11. The process of claim 8 , wherein culturing or fermenting is performed using a batch process.
12. The process of claim 8 , wherein culturing or fermenting is performed using a fed batch process.
13. The process of claim 8 , wherein culturing or fermenting is performed using a repeated fed batch process.