Properties of a Purified Halophilic Malic Dehydrogenase

Abstract

Holmes, P. K. (University of Illinois, Urbana), and H. Orin Halvorson . Properties of a purified halophilic malic dehydrogenase. J. Bacteriol. 90: 316–326. 1965.—The malic dehydrogenase (MDH) from Halobacterium salinarium required high concentrations of monovalent ions for stability and activity. Studies of inactivation rates at different salt concentrations suggested that approximately 25% NaCl (w/v) is required to stabilize MDH. From 50 to 100% reactivation, depending on the salt concentration present during inactivation, could occur in 2.5 to 5 m NaCl or KCl. The optimal salt concentration for activity of MDH was a function of the p H, and ranged from 1 to 3 m NaCl or KCl. The effect of salt concentration on the p H-activity curves occurred chiefly below p H 7.0. Inactivation of MDH with heat or thiol reagents showed that the enzyme was more labile in the state induced by absence of salt. The activation of MDH by salts was attributed to a decreased rate of dissociation of MDH and reduced nicotinamide adenine dinucleotide (NADH 2 ). The inactivation of the enzyme in the absence of salt could be largely prevented by the presence of NADH 2 . The S 20.w of MDH decreased threefold at low salt concentrations. The enzyme was assumed to be in its native compact configuration only in the presence of a high concentration of salt.