Study of umbelliferone hydroxylation to esculetin catalyzed by polyphenol oxidase.

We characterize umbelliferone, a derivative of 2,4-dihydroxycoumaric acid, as a substrate of polyphenol oxidase. This enzyme hydroxylates umbelliferone to esculetin, its o-diphenol, and then oxidizes it to o-quinone. The findings show that umbelliferone, an intermediate in one of the coumarin biosynthesis pathways, may be transformed into its o-diphenol, esculetin, which is also an intermediate in the same pathway.

Effects of tetrahydropterines on the generation of quinones catalyzed by tyrosinase.

Tetrahydrobiopterine (6BH(4)) can diminish the oxidative stress undergone by keratinocytes and melanocytes by reducing the o-quinones generated by the oxidation of the corresponding o-diphenols. We found that 6BH(4) and their analogs reduced all the o-quinones studied. The formal potentials of different quinone/diphenol pairs indicate that the o-quinones with withdrawing groups are more potent oxidants than those with donating groups.

Kinetic characterization of the oxidation of carbidopa and benserazide by tyrosinase and peroxidase.

Carbidopa and benserazide have been described as inhibitors of dopa decarboxylase and both have been used in the treatment of Parkinson's disease. Because of their chemical structure as polyphenols, these compounds can behave as substrates of tyrosinase and peroxidase. We demonstrate that these enzymes oxidize both substrates.

Kinetic characterization of the oxidation of esculetin by polyphenol oxidase and peroxidase.

Esculetin has been described as an inhibitor of tyrosinase and polyphenol oxidase and, therefore, of melanogenesis. In this work, we demonstrate that esculetin is not an inhibitor but a substrate of mushroom polyphenol oxidase (PPO) and horseradish peroxidase (POD), enzymes which oxidize esculetin, generating its o-quinone. Since o-quinones are very unstable, the usual way of determining the enzymatic activity (slope of recordings) is difficult.