Identification by electron spin resonance of free radicals formed during the oxidation of 4-hydroxyanisole catalyzed by tyrosinase.

We have observed the formation of free radicals during the oxidation of the melanocytotoxic agent 4-hydroxyanisole with the enzyme tyrosinase as a catalyst. The first free radical to form is identified as the 4-methoxy-1,2-benzosemiquinone radical anion. The peak concentration of this radical increases with tyrosinase concentration; a minimum concentration of 50 micrograms/ml of tyrosinase was needed to observe this radical. The peak concentration of this radical is independent of 4-hydroxyanisole concentration. This radical is produced by reverse dismutation of the primary product, 4-methoxy-1,2-benzoquinone and 4-methoxycatechol produced indirectly.