title: Properties of aspartate racemase, a pyridoxal 5'-phosphate-independent amino acid racemase.
abstract: Aspartate racemase from Streptococcus thermophilus contains no pyridoxal 5'-phosphate or other cofactors such as FAD, NAD+, and metal ions. It was affected by neither carbonyl reagents such as hydroxylamine nor sodium borohydride but was strongly inhibited by iodoacetamide and other thiol reagents. Aspartate, cysteate, and cysteine sulfinate were the only substrates. The Km values for L- and D-aspartate were 35 and 8.7 mM, respectively. The enzyme catalyzed the exchange of alpha-hydrogen of the substrate with the solvent hydrogen. Racemization of L-aspartate in 2H2O showed an overshooting in the optical rotation of aspartate before the substrate was fully racemized. This shows that the removal of alpha-hydrogen of the substrate is at least partially rate-determining. When L- or D-aspartate was incubated with aspartate racemase in tritiated water, tritium was incorporated preferentially into the product enantiomer. The results strongly suggest that aspartate racemase contains two hydrogen acceptors.
authors.affiliation: Institute for Chemical Research, Kyoto University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Yamauchi
authors.forename: S Y
authors.initials: SY
authors.lastname: Choi
authors.forename: H
authors.initials: H
authors.lastname: Okada
authors.forename: M
authors.initials: M
authors.lastname: Yohda
authors.forename: H
authors.initials: H
authors.lastname: Kumagai
authors.forename: N
authors.initials: N
authors.lastname: Esaki
authors.forename: K
authors.initials: K
authors.lastname: Soda
mesh_terms: D000591:Amino Acid Isomerases / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D008670:Metals / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000378:metabolism*; D013291:Streptococcus / Q000201:enzymology; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds
chemical_list: D008670:Metals; D013438:Sulfhydryl Compounds; D009243:NAD; D005182:Flavin-Adenine Dinucleotide; D011732:Pyridoxal Phosphate; D000591:Amino Acid Isomerases; C019749:aspartate racemase