ABILITY OF VARIOUS CHEMICALS TO REDUCE COPPER AND TO INACTIVATE MUSHROOM TYROSINASE1

Treatment of mushroom tyrosinase with reducing agents such as hydrogen peroxide, ascorbic acid, phenylhydrazine, gallic acid, ferrocyanide and NH2OH resulted in inactivation of the enzyme. Under the conditions tested, 50% inactivation of the enzyme was obtained with 4 μM H2O2, 20 μM ascorbic acid, 40μM phenylhydrazine, 6 mM gallic acid, 12 mM ferrocyanide and 22 mM NH2OH. The ability of the reducing agents to reduce Cu2+ in a chemical model system was determined and it was found that gallic acid, phenylhydrazine, ascorbic acid and NH2OH are relatively good reductants of Cu2+ while H2O2 and ferrocyanide are relatively poor ones. The copper content of mushroom tyrosinase before and after inactivation by each of the reducing agents was determined. The copper content of the enzyme inactivated by H2O2, NH2OH, phenylhydrazine, ferrocyanide, gallic acid and ascorbic acid was 100%, 90%, 90%, 85%, 85% and 76% compared to that of the control (enzyme not treated). It was concluded that the degree of inactivation of mushroom tyrosinase by the reducing agents was not correlated with the decrease in the copper content of the enzyme nor with their ability to reduce Cu2+ in a chemical model system.