Purification of an NADH-(dichlorophenol-indophenol) oxidoreductase from Bacillus stearothermophilus.
An NADH-(dichlorophenol-indophenol) oxidoreductase was purified 104-fold and in 25% overall yield from the thermophilic bacterium Bacillus stearothermophilus, strain PH24. After solubilization in 2M-NaCl at 70 degrees C, the enzyme was purified by ion-exchange and hydroxyapatite chromatography, followed by affinity chromatography on immobilized Cibacron Blue 3GA. The purified enzyme had a mol.wt. of 43 000 and had an absorption spectrum characteristic of flavoprotein. The enzyme activity was enhanced by FMN and by CN-. The enzyme was inhibited by EDTA and by rho-chloromercuribenzoic acid.
D015086:2,6-Dichloroindophenol; D002853:Chromatography, Liquid; D003579:Cytochrome Reductases / Q000302:isolation & purification*; D004789:Enzyme Activation / Q000187:drug effects; D005486:Flavin Mononucleotide / Q000494:pharmacology; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D008722:Methods; D009245:NADH Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D011190:Potassium Cyanide / Q000494:pharmacology
D005486:Flavin Mononucleotide; D015086:2,6-Dichloroindophenol; D003579:Cytochrome Reductases; D009245:NADH Dehydrogenase; D011190:Potassium Cyanide