Method for increasing the metabolic flux through the pentose phosphate cycle in coryneform bacteria by regulation of the phosphoglucose isomerase (pgi gene)
AssigneeDegussa AG
InventorsL. K. Dunican, Ashling McCormack, Cliona Stapleton, Kevin Burke, Michael O'Donohue, Achim Marx, Bettina Mockel
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Abstract
The present invention is directed to isolated polynucleotides coding for phosphoglucose isomerase (pgi) from coryneform bacteria. In addition, the invention includes methods for increasing the metabolic flux through pentose phosphate cycle of bacteria by reducing or eliminating the activity of pgi. These methods may be used to increase the fermentative production of nucleotides, vitamins and amino acids.
Claims8
A process for increasing the metabolic flux through the pentose phosphate cycle in coryneform bacteria, said process comprising:
- (a) eliminating the intracellular activity of a phosphoglucose isomerase (pgi) enzyme comprising the amino acid sequence of SEQ ID NO:2 in said coryneform bacteria;
- and
- (b) fermenting the coryneform bacteria of step (a), wherein elimination is achieved by one or more methods of mutagenesis selected from the group consisting of deletion mutagenesis with deletion of two or more codons in the pgi gene;
- insertional mutagenesis due to homologous recombination;
- and transition or transversion mutagenesis with incorporation of a non-sense mutation in the pgi gene.
A process for producing L-lysine using coryneform bacteria, said process comprising:
- (a) eliminating the intracellular activity of a phosphoglucose isomerase (pgi) enzyme comprising the amino acid sequence of SEQ ID NO:2 in said coryneform bacteria;
- (b) fermenting the coryneform bacteria of step (a);
- (c) allowing L-lysine to accumulate in the fermentation medium or in said coryneform bacteria of step (b);
- and
- (d) isolating said L-lysine, wherein elimination is achieved by one or more methods of mutagenesis selected from the group consisting of deletion mutagenesis with deletion of two or more codons in the pgi gene;
- insertional mutagenesis due to homologous recombination;
- and transition or transversion mutagenesis with incorporation of a non-sense mutation in the pgi gene.
A process for increasing the metabolic flux through the pentose phosphate cycle in bacteria of the species Corynebacterium glutamicum, said process comprising:
- (a) eliminating the intracellular activity of the endogenous phosphoglucose isomerase (pgi) enzyme in said bacteria;
- and
- (b) fermenting the bacteria of step (a), wherein elimination is achieved by one or more methods of mutagenesis selected from the group consisting of deletion mutagenesis with deletion of two or more codons in the pgi gene;
- insertional mutagenesis due to homologous recombination;
- and transition or transversion mutagenesis with incorporation of a non-sense mutation in the pgi gene.
A process for producing L-lysine using bacteria of the species Corynebacterium glutamicum, said process comprising:
- (a) eliminating the intracellular activity of the endogenous phosphoglucose isomerase (pgi) enzyme in said bacteria;
- (b) fermenting the bacteria of step (a);
- (c) allowing said L-lysine to accumulate in the fermentation medium or in said bacteria of step (b);
- and
- (d) isolating L-lysine, wherein elimination is achieved by one or more methods of mutagenesis selected from the group consisting of deletion mutagenesis with deletion of two or more codons in the pgi gene;
- insertional mutagenesis due to homologous recombination;
- and transition or transversion mutagenesis with incorporation of a non-sense mutation in the pgi gene.
Dependencies are inferred from the claim text.