Molecular cloning, sequence and characterization of a novel streptococcal phosphoglycerate dehydrogenase gene.
The nucleotide sequence of a Streptococcus mutans serA gene that encodes D-3-phosphoglycerate dehydrogenase has been determined. The gene consisted of 1308-bp nucleotides coding for a 436-amino-acid polypeptide (48,546 Da). The deduced amino acid sequence showed a 66% identity with SerA from Bacillus subtilis and possessed specific residues (G-R-P-N-V-G) in the coenzyme-binding domain, alpha B helix. Recombinant streptococcal SerA was expressed using pMAL-c2 expression vector and purified by amylose resin affinity chromatography and DEAE-Sephacel column chromatography. This SerA enzyme catalyzed detectable reduction of alpha-ketoglutarate to 2-hydroxyglutaric acid. These findings indicate that the novel streptococcal phosphoglycerate dehydrogenase, SerA, is a member of a D-isomer-specific family of 2-hydroxyacid dehydrogenases.
Kawabata
S
S
Department of Oral Microbiology, Osaka University Faculty of Dentistry, Suita-Osaka, 565-0871 Japan.
Terao
Y
Y
Hamada
S
S
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D005810:Multigene Family; D050543:Phosphoglycerate Dehydrogenase; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017385:Sequence Homology; D013295:Streptococcus mutans / Q000201:enzymology* / Q000235:genetics*
D001426:Bacterial Proteins; D002237:Carbohydrate Dehydrogenases; D050543:Phosphoglycerate Dehydrogenase