Some properties of an alcohol dehydrogenase partially purified from baker's yeast grown without added zinc.
Alcohol dehydrogenase was partially purified from yeast (Saccharomyces cerevisiae) grown in the presence of 20 muM-MnSO4 without added Zn2+ and from yeast grown in the presence of 1.8 muM-MnSO4. The enzyme from yeast grown with added Zn2+ has the same properties as the crystalline enzyme from commercial supplies of baker's yeast. The enzyme from yeast grown without added An2+ has quite different properties. It has a mol.wt. in the region of 72000 and an S 20 w of 5.8S. The values can be compared with a mol.wt. of 141000 and an S 20 w of 7.6S for the crystalline enzyme. ADP-ribose, a common impurity in commercial samples of NAD+, is a potent competitive inhibitor of the new enzyme (K1 = 0.5 muM), but is not so for the crystalline enzyme. The observed maximum rate of ethanol oxidation at pH 7.05 and 25 degrees C was decreased 12-fold by the presence of 0.06 mol of inhibitor/mol of NAD+ when using the enzyme from Zn2+-deficient yeast, but with crystalline enzyme the maximum rate was essentially unchanged by this concentration of inhibitor. The kinetic characteristics for the two enzymes with ethanol, butan-1-ol, acetaldehyde and butyraldehyde as substrates are markedly different. These kinetic differences are discussed in relation to the mechanism of catalysis for the enzyme from Zn2+-deficient yeast.
D000079:Acetaldehyde; D000247:Adenosine Diphosphate Sugars / Q000494:pharmacology; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000447:Aldehydes; D000440:Butanols; D000431:Ethanol; D007700:Kinetics; D008345:Manganese / Q000032:analysis; D008970:Molecular Weight; D009243:NAD; D012441:Saccharomyces cerevisiae / Q000032:analysis / Q000201:enzymology*; D015032:Zinc / Q000032:analysis / Q000378:metabolism*
D000247:Adenosine Diphosphate Sugars; D000447:Aldehydes; D000440:Butanols; D009243:NAD; D000431:Ethanol; D008345:Manganese; D000429:Alcohol Oxidoreductases; D000079:Acetaldehyde; D015032:Zinc