Purification and Kinetic Studies of Yeast Alcohol Dehydrogenase I.
An efficient procedure for the purification of yeast alcohol dehydrogenase I (ADHI) was developed. By using Blue-Sepharose 4B affinity chromatography, ADHI from yeast (S. cerevisiae) was purified 200-fold with an overall yield of 47% to homogeneity as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis and starch gel electrophoresis. Results of studies on the product and dead-end inhibition kinetics were in agreement with and ordered Bi Bi mechanism as proposed by Wratten and Cleland. Dioxane was found to be a competitive inhibitor of this enzyme. The complexity of the reaction mechanism of this enzyme is discussed.
Lian
De-Jun
DJ
Shanghai Institute of biochemistry, Academia Sinica, Shanghai 200031, China.
Li
Lin
L
Xu
Gen-Jun
GJ