Abstract
As a fundamental study to develop an efficient method for the separation of α- and β-naphthols by combining enzymatic reaction and selective extraction, we investigated the kinetics and the mechanism of the hydrolysis of α-and β-naphthyl sulfates catalyzed by free sulfatase from Helix pomatia. These hydrolysis reactions occur competitively and obey a Michaelis-Menten type mechanism. The products, α- and β-naphthols, are inhibitors of both hydrolysis reactions. The enzyme was immobilized in porous gel lattices of albumin and glutaraldehyde. We measured the initial rates of the hydrolysis reactions catalyzed by the membrane-bound sulfatase and analyzed them in terms of the catalytic effectiveness factor. The kinetic parameters evaluated were almost the same as those for free enzyme, except that Km for α-naphthyl sulfate and kB/kA the ratio of the production rate constants were respectively 1/6.1 and 1/2.4 those of free enzyme.
