3-Hydroxy-3-methylglutaryl coenzyme a reductase from the latex of Hevea brasiliensis

Abstract

3-Hydroxy-3-methylglutaryl coenzyme A reductase (mevalonate: NADP + oxidoreductase acylating CoA; EC 1.1.1.34) was purified from fresh Hevea latex of clone RRIM 600. The latex was centrifuged and the sediment used for enzyme purification. The enzyme was solubilized by freeze-thawing in a buffer containing 1 % Brij W-1 and 20% glycerol. Affinity chromatography (HMG-CoA-Hexyl-Agarose) was used in the final purification step. The M r determined by SDS-PAGE was 44000 and that estimated from non-denaturing gel electrophoresis, 176000. The optimal pH was ca 7, with an apparent K m of 13 μM for (S)-HMG-CoA. A low concentration of dithiothreitol was required for maximal activity of the purified enzyme. © 1990.