Chemical and enzymatic oxidation of 4-methylcatechol in the presence and absence of L-serine. Spectrophotometric determination of intermediates.

A pathway for 4-methylcatechol oxidation by tyrosinase (monophenol, dihydroxy-L-phenylalanine:oxygen oxidoreductase, EC 1.14.18.1) in the presence of L-serine is proposed. Characterization of intermediates in this oxidative reaction and stoichiometry determination have both been performed. It has been possible to detect spectrophotometrically 4-methyl-o-benzoquinone as the first intermediate in this pathway after oxidizing 4-methylcatechol with epidermis tyrosinase or sodium periodate at pH 6.5. The steps for 4-methylcatechol transformation in the presence of L-serine could be: 4-methylcatechol----4-methyl-o-benzoquinone----5-methyl-4N-serine-catechol----5 - methyl-4N-serine-o-benzoquinone. Matrix analysis of the spectra obtained with a rapid-scan spectrophotometer verified that 4-methyl-o-benzoquinone was transformed into 5-methyl-4N-serine-o-benzoquinone at a constant ratio, the stoichiometry for this conversion being the equation: 2-(4-methyl-o-benzoquinone) + L-serine----4-methylcatechol + 5-methyl-4N-serine-o-benzoquinone.