Purification and some of the properties of a novel secondary alcohol dehydrogenase from Alcaligenes eutrophus

Abstract

Alcaligenes eutrophus utilizing nerolidol, a sesquiterpene alcohol,as the sole source of carbon contains an inducible NAD(P)⁺-linked secondary alcohol dehydrogenase (SADH). The enzyme was purified to homogeneity by a combination of salt precipitation, ion exchange and affinity matrix chromatographies. The apparent molecular mass of the enzyme was estimated to be 139 KDa with four identical subunits of 38.5 KDa. The enzyme carried out both oxidation and reduction reactions. At pH 5.5, enzyme catalyzed the stereospecific reduction of prochiral ketones to secondary alcohols. The pH optimum for the oxidation reaction was 9.5. NADP⁺ and NADPH were respectively preferred over NAD⁺ and NADH for oxidation and reduction reactions. Some of the properties of this enzyme were found to be significantly different from those thus far described.