Collect. Czech. Chem. Commun. 1981, 46, 798-806

Purification and characterization of alcohol dehydrogenase from pig liver

Radim Pařízek, Jan Kovář and Ladislav Skurský

Department of Biochemistry, Purkyně University, 611 37 Brno


Alcohol dehydrogenase from pig liver was purified from a tissue homogenate under conditions of enzyme protection in a complex with NADH and isobutyramide and further by chromatography on ion-exchange celluloses and Sephadex G-100. pH 8 and an addition of dithiotreitol was found most effective for stabilization of the enzyme. Electrophoretic behavior revealed the existence of at least 4 isoenzymes with variable relative amounts of 3 minor components. Using a titration of the enzyme by NADH in the presence of isobutyramide the concentration of active sites and turnover number of the enzyme were determined. The basic kinetic characteristics show that the pig enzyme is less active than that of the horse origin, whereas its affinity to coenzymes and substrates is comparable. Pig alcohol dehydrogenase can be inhibited by inhibitors active against other animal alcohol dehydrogenases.