Purification and properties of alpha-ketoglutarate reductase from Micrococcus aerogenes.
Micrococcus aerogenes grown in media containing glutamate has high levels of glutamate dehydrogenase and alpha-ketoglutarate reductase. The latter enzyme catalyzes the reversible reduction of alpha-ketoglutarate to alpha-hydroxyglutarate in the presence of reduced nicotinamide adenine dinucleotide (NADH). The enzyme has a high specificity for both substrates in either direction and displays Michaelis-Menten kinetics at moderate substrate concentrations. K(m) values of 0.12 to 0.17 mm alpha-ketoglutarate and 0.3 mm NADH for the forward reaction were calculated from data obtained at low substrate concentrations. At high concentrations, this reaction was inhibited by both substrates. The reverse reaction, which proceeded at 0.1 to 0.2 times the rate of the forward reactions, was inhibited by one of the products, alpha-ketoglutarate. K(m) values for the substrates of this reaction were 10 mm for alpha-hydroxyglutarate and 1 mm for nicotinamide adenine dinucleotide. alpha-Ketoglutarate reductase has a molecular weight of 7.5 x 10(4) to 8.2 x 10(4) and is composed of identical polypeptide chains with a molecular weight of 3.6 x 10(4) to 3.8 x 10(4).
D000179:Acrylates; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002854:Chromatography, Paper; D003470:Culture Media; D004586:Electrophoresis; D005782:Gels; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D005977:Glutarates / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D006882:Hydroxyapatites; D007656:Ketoglutaric Acids / Q000378:metabolism; D008837:Micrococcus / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D000644:Quaternary Ammonium Compounds; D013045:Species Specificity; D013053:Spectrophotometry; D013395:Sucrose; D013431:Sulfates
D000179:Acrylates; D003470:Culture Media; D005782:Gels; D005971:Glutamates; D005977:Glutarates; D006882:Hydroxyapatites; D007656:Ketoglutaric Acids; D000644:Quaternary Ammonium Compounds; D013431:Sulfates; D009243:NAD; D009249:NADP; D013395:Sucrose; D000429:Alcohol Oxidoreductases; D005969:Glutamate Dehydrogenase