Soluble reduced nicotinamide adenine dinucleotide oxidase from Bacillus cereus T spores and vegetative cells. I. Purification

Abstract

Spores and vegetative cells of Bacillus cereus T were disrupted by two procedures and soluble extracts prepared from the ruptured cells. Reduced nicotinamide adenine dinucleotide (NADH2) oxidases were purified from the extracts by ammonium sulfate fractionation, ion exchange on hydroxylapatite, and preparative acrylamide gel electrophoresis. The electrophoresis step revealed the presence of two distinct components with NADH2 oxidase activity in soluble extracts of spores while vegetative cell extracts contained only one. The faster moving component in spore extracts constituted about 30% of the NADH2 oxidase activity recovered and was identical with the vegetative cell enzyme in electrophoretic mobility on acrylamide gel. The slower moving spore component accounted for 70% of recovered activity and was found in soluble extracts regardless of the procedure used to rupture spores.