Nucleotide sequences that encode deregulated phosphoglycerate dehydrogenases of coryneform bacteria and method for producing L-serine
AssigneesForschungszentrum Julich GmbH, Amino Corporation
InventorsLothar Eggeling, Petra Peters-Wendisch, Roman Netzer, Hermann Sahm, Robert Faurie, Birgit Klassen
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Abstract
The invention relates to nucleotide sequences of coryneform bacteria that encode proteins that are involved in the biosynthesis of L-serine and to a method for producing L-serine. According to the invention, at least 79 amino acids at the C terminus of the wild-type serA sequence are deleted, thereby producing a 3-phosphoglycerate dehydrogenase having a reduced feedback inhibition by L-serine vis-à-vis the wild-type sequence.
Claims28
An isolated nucleic acid consisting of SEQ ID NO:
- 1 where said sequence encodes a deregulated 3-phosphoglycerate dehydrogenase, which in comparison to a wild type 3-phosphoglycerate dehydrogenase has reduced feedback inhibition by L-serine where said nucleic acid is a fragment of an SerA gene.
An isolated nucleic acid consisting of SEQ ID NO:
- 2 where said sequence encodes a deregulated 3-phosphoglycerate dehydrogenase, which in comparison to a wild type 3-phosphoglycerate dehydrogenase has reduced feedback inhibition by L-serine where said nucleic acid is a fraqment of an SerA gene.
An isolated nucleic acid consisting of SEQ ID NO:
- 3 where said sequence encodes a deregulated 3-phosphoglycerate dehydrogenase, which in comparison to a wild type 3-phosphoglycerate dehydrogenase has reduced feedback inhibition by L-serine where said nucleic acid is a fraqment of an SerA gene.
An isolated nucleic acid consisting of SEQ ID NO:
- 4 where said sequence encodes a deregulated 3-phosphoglycerate dehydrogenase, which in comparison to a wild type 3-phosphoglycerate dehydrogenase has reduced feedback inhibition by L-serine where said nucleic acid is a fraqment of an SerA gene.
An isolated nucleic acid consisting of SEQ ID NO:
- 5 where said sequence encodes a deregulated 3-phosphoglycerate dehydrogenase, which in comparison to a wild type 3-phosphoglycerate dehydrogenase has reduced feedback inhibition by L-serine where said nucleic acid is a fraqment of an SerA gene.
A mutant deregulated 3-phosphoglycerate dehydrogenase where said mutant 3-phosphoglycerate dehydrogenase is expressed from a nucleic acid consisting of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4 or SEQ ID NO:
- 5 that encode the amino acid sequence of SEQ ID NO:
- 7, SEQ ID NO:
- 8, SEQ ID NO:
- 9, SEQ ID NO:
- 10 or SEQ ID NO:
- 11 respectively and where said mutant deregulated 3-phosphoglycerate-dehydrogenase has a reduced feedback inhibition by L-serine compared to the wild type 3-phosphoglycerate dehydrogenase.
A probe for identifying and/or isolating genes which encode a deregulated 3-phosphoglycerate dehydrogenase participating in the biosynthesis of L-serine, said probe consisting of SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, SEQ ID NO. 18, or SEQ ID NO.19 and containing a marker suitable for detection.
A method for microbially producing L-serine from a carbohydrate, fat or oil, fatty acid, alcohol or organic acid, in a culture medium, containing nitrogen sources and phosphorous sources, which comprises the steps of:
- a) providing at least one nucleic acid encoding a deregulated 3-phosphoglycerate dehydrogenase, selected from the group consisting of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4 and SEQ ID NO. 5, isolated from a Coryneform bacterium, and transformed into a Coryneform bacterium, and then expressed to form the deregulated 3-phosphoglycerate dehydrogenase, whereby the gene expression and/or the activity of the corresponding encoded deregulated 3-phosphoglycerate dehydrogenase is increased with respect to the corresponding microorganism which has not been genetically altered;
- b) microbially producing L-serine by expressing the at least one nucleic acid which encodes a deregulated 3-phosphoglycerate dehydrogenase in said genetically modified microorganism from step a) to microbially convert said carbohydrate, fat or oil, fatty acid, alcohol or organic acid in said culture medium to L-serine;
- and
- c) isolating the correspondingly formed L-serine from the culture medium.
Dependencies are inferred from the claim text.