PURIFICATION OF ALCOHOL-DEHYDROGENASE FROM ENTAMOEBA-HISTOLYTICA AND SACCHAROMYCES-CEREVISIAE USING ZINC-AFFINITY CHROMATOGRAPHY

We have developed a single-step method for the purification of NADP(+) -dependent alcohol dehydrogenase from Entamoeba histolytica and NAD(+) -dependent alcohol dehydrogenase from Saccharomyces cerevisiae. It is based oil the affinity for zinc of both enzymes. The amebic enzyme was purified almost 800 times with a recovery of 54% and the yeast enzyme was purified 30 times with a recovery of 100%. The kinetic constants of the purified enzymes were similar to those reported for other purif ication methods. With mammalian alcohol dehydrogenase, we obtained a 4 0-kDa band suggestive of purified alcohol dehydrogenase, but we failed to retain enzymatic activity in this preparation. Our results suggest that the described method is more applicable to the purification of t etrameric alcohol dehydrogenase. (C) 1997 Academic Press.