N. Cabrera et al., PURIFICATION OF ALCOHOL-DEHYDROGENASE FROM ENTAMOEBA-HISTOLYTICA AND SACCHAROMYCES-CEREVISIAE USING ZINC-AFFINITY CHROMATOGRAPHY, Protein expression and purification, 10(3), 1997, pp. 340-344
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.