Purification and properties of Lactobacillus plantarum inducible malic enzyme.
Inducible malic enzyme (l-malate:NAD oxidoreductase [decarboxylating], EC 1.1.1.39) was isolated from Lactobacillus plantarum and purified about 100-fold with 27% yield of the original activity. Kinetic studies with the purified malic enzyme yielded the following results: pH optimum, 7.6 to 8.0; K(m) for l-malate, 0.38 mM; K(m) for NAD, 0.072 mM; and K(m) for MnCl(2), 0.048 mM. It was shown that this enzyme was inhibited by high concentrations of substrate and nicotinamide adenine dinucleotide (NAD), indicating it may be regulated by substrate or NAD. Molecular weight of 130,000 +/- 10,000 was determined by Sephadex gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point, determined by isoelectrofocusing, was 4.3 at 7 C. Isoelectrofocusing also resolved three active peaks which focused at pH 4.19, 4.31, and 4.40.
D002474:Cell-Free System; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D005612:Freeze Drying; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007778:Lactobacillus / Q000201:enzymology*; D008291:Malate Dehydrogenase / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D008298:Maleates / Q000378:metabolism; D008365:Manometry; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D013053:Spectrophotometry; D013237:Stereoisomerism
D008298:Maleates; D009243:NAD; D008291:Malate Dehydrogenase