title: A new family of D-2-hydroxyacid dehydrogenases that comprises D-mandelate dehydrogenases and 2-ketopantoate reductases.
abstract: The gene for the D-mandelate dehydrogenase (D-ManDH) of Enterococcus faecalis IAM10071 was isolated by means of an activity staining procedure and PCR and expressed in Escherichia coli cells. The recombinant enzyme exhibited high catalytic activity toward various 2-ketoacid substrates with bulky hydrophobic side chains, particularly C3-branched substrates such as benzoylformate and 2-ketoisovalerate, and strict coenzyme specificity for NADH and NAD(+). It showed marked sequence similarity with known NADP-dependent 2-ketopantoate reductases (KPR). These results indicate that together with KPR, D-ManDH constitutes a new family of D-2-hydroxyacid dehydrogenases that act on C3-branched 2-ketoacid substrates with various specificities for coenzymes and substrates.
authors.affiliation: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Wada
authors.forename: Saho
authors.initials: S
authors.lastname: Iwai
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Tamura
authors.forename: Tomonori
authors.initials: T
authors.lastname: Ando
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shinoda
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Arai
authors.forename: Hayao
authors.initials: H
authors.lastname: Taguchi
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D013194:Staining and Labeling
chemical_list: D000429:Alcohol Oxidoreductases; C043630:D-mandelate dehydrogenase; C021934:2-dehydropantoate 2-reductase; C021379:D-2-hydroxyacid dehydrogenase