1. A new family of D-2-hydroxyacid dehydrogenases that comprises D-mandelate dehydrogenases and 2-ketopantoate reductases.
- Author
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Wada Y, Iwai S, Tamura Y, Ando T, Shinoda T, Arai K, and Taguchi H
- Subjects
- Alcohol Oxidoreductases chemistry, Alcohol Oxidoreductases genetics, Amino Acid Sequence, Base Sequence, Electrophoresis, Polyacrylamide Gel, Enterococcus faecalis genetics, Gene Expression Regulation, Bacterial, Kinetics, Molecular Sequence Data, Polymerase Chain Reaction, Sequence Alignment, Staining and Labeling, Alcohol Oxidoreductases metabolism, Enterococcus faecalis enzymology
- 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.
- Published
- 2008
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