A highly specific glyoxylate reductase derived from a formate dehydrogenase.
A Glu141Asn mutant Paracoccus sp. 12-A formate dehydrogenase catalyzes marked glyoxylate reduction. Additional replacement of the His332-Gln313 pair with His-Glu, which is a consensus acid/base catalyst in D-hydroxyacid dehydrogenases, further improved the catalytic activity of the enzyme as to glyoxylate reduction through enhancement of the hydrogen transfer step in the catalytic process, slightly shifting the optimal pH for the reaction. On the other hand, the replacement induced no marked activity toward other 2-ketoacid substrates, and diminished the enzyme activity as to formate oxidation. Consequently, the formate dehydrogenase was converted to a highly specific and active glyoxylate reductase through only the two amino acid replacements.
Shinoda
Takeshi
T
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Arai
Kazuhito
K
Taguchi
Hayao
H
D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D019943:Amino Acid Substitution; D002384:Catalysis; D020134:Catalytic Domain / Q000235:genetics*; D005560:Formate Dehydrogenases / Q000737:chemistry / Q000235:genetics*; D006038:Glyoxylates / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D010084:Oxidation-Reduction; D010230:Paracoccus / Q000201:enzymology* / Q000235:genetics
D006038:Glyoxylates; D000429:Alcohol Oxidoreductases; C020830:glyoxylate reductase; D005560:Formate Dehydrogenases; C031150:glyoxylic acid