Hydrated electron-induced inactivation of tyrosinase in aqueous solution by exposure to cobalt-60 gamma-rays. II. Catecholase activity

Tyrosinase (0.2 mg/ml) was irradiated with ⁶⁰Co gamma-rays. The catecholase activity was measured at varying radiation doses under various atmospheric conditions. Do was found to be 1.25 kGy and hit number to be 2 in N₂-saturated solution. OH radical scavengers t@?-BuOH and MeOH, had no effect. O₂ which is an enhancer of OH-induced enzyme inactivation had little effect. But N₂O as a e(aq) scavenger and Cu⁺⁺ markedly protected against the inactivation indicating that e(aq) was the main species to inactivate the enzymatic activity. By Ultrogel chromatography, it was found that the enzymatic activity was lost when this enzyme dissociated into its subunits. Thus, it was concluded that the radiation-induced inactivation was due to the reduction of Cu⁺⁺ as the active center and the chelater with e(aq) followed by the dissociation.