PURIFICATION AND PROPERTIES OF BENZYLALCOHOL DEHYDROGENASE FROM PSEUDOMONAS SP

An NAD-linked alcohol dehydrogenase (Alcohol:NAD oxidoreductase, EC 1.1.1.1.), which catalyzes the interconversion of benzylalcohol and benzaldehyde has been isolated from Pseudomonas putida T-2 grown on toluene as sole source of carbon and energy. The purified enzyme was shown to be homogeneous by sedimentation in the ultracentrifuge, and by disc gel electrophoresis. The molecular weight was approximately 110,000. The enzyme was unstable with a half-life of few hours under conventional conditions. The instability was overcome by using the acetone buffer. The enzyme was also protected effectively by ammonium or sodium sulfates. The purified preparation of the enzyme reacted with a range of primary alcohols with aromatic and cyclohexene-1 ring, but not with aliphatic alcohols such as ethanol although it showed only slight activity toward C6-C8 aliphatic normal alcohols. Enzymatic activity was extremely sensitive to sulfhydryl reagents such as p-chloromercuribenzoate and N-ethylmaleimide. © 1969.