Studies on methyl chloride dehalogenase and O-demethylase in cell extracts of the homoacetogen strain MC based on a newly developed coupled enzyme assay

摘要: An enzyme assay was developed to determine the activities of methyl chloride dehalogenase and O-demethylase homoacetogen strain MC. The formation tetrahydrofolate from or vanillate coupled oxidation methylene mediated by reductase purified Peptostreptococcus productus (strain Marburg) subsequent methenyl catalyzed dehydrogenase same organism. To drive endergonic with NAD+ as an electron acceptor, NADH formed in this reaction reoxidized exergonic lactate reaction. NADPH followed photometrically at 350 nm; e350 about 29.5 mM–1cm–1 (pH 6.5). Using assay, cofactor requirements, apparent kinetic parameters, pH temperature optima both enzymes, effect inhibitors were determined. activity dependent on presence ATP; arsenate severely inhibited absence ATP. described allows purification characterization is also appropriate for enzymatic determination tetrahydrofolate.