The effect of catalase on the inactivation of tyrosinase by ascorbic acid and by cysteine or glutathione

Abstract

When tyrosine was incubated with tyrosinase in the presence of ascorbic acid, dopa and 5-hydroxydopa were formed and the enzyme was inactivated. In the presence of catalase, more dopa and 5-hydroxydopa were formed because enzyme inactivation was prevented. Incubation of dopa and cysteine with small amounts of mushroom tyrosinase led to rapid inactivation of the enzyme. This inactivation was accelerated in the presence of catalase. New systems developed have been useful in demonstrating the role of hydrogen peroxide in tyrosinase inactivation by several compounds of importance in melanin biochemistry. Cysteine and glutathione inactivated tyrosinase. Addition of catalase increased the inactivation at high thiol concentrations, but decreased the inactivation at low concentrations. Ascorbic acid and 5-hydroxydopamine also inactivated tyrosinase, but with these compounds inactivation was completely prevented by addition of catalase. The inactivation by dopamine was negligible under the experimental conditions. Inactivation of tyrosinase by ascorbic acid and by 5-OH-dopamine was found dependent on oxygen, whereas inactivation by cysteine and glutathione was independent of oxygen. Large amounts of serum albumin protected tyrosinase from inactivation by ascorbic acid and 5-OH-dopamine, but did not prevent inactivation by cysteine and glutathione. The presence of substrate had a protective effect on the inactivation of tyrosinase by cysteine.