The catalytic cycle of tyrosinase: peroxide attack on the phenolate ring followed by O-O bond cleavage

The oxidation of phenols to ortho-quinones, catalyzed by tyrosinase, has been studied using the hybrid DFT method B3LYP. Since no X-ray structure exists for tyrosinase, information from the related enzymes hemocyanin and catechol oxidase were used to set up a chemical model for the calculations. Previous studies have indicated that the direct cleavage of O2 forming a Cu2(III,III) state is energetically very unlikely. The present study therefore followed another mechanism previously suggested. In this mechanism, dioxygen attacks the phenolate ring which is then followed by O-O bond cleavage. The calculations give a reasonable barrier for the O2 attack of only 12.3 kcal/mol, provided one of the copper ligands is able to move substantially away from its direct copper coordination. This can be achieved with six histidine ligands even if these ligands are held in their positions by the enzyme, but can also be achieved if one of the coppers only has two histidine ligands and the third ligand is water. The next step of O-O bond cleavage has a computed barrier of 14.4 kcal/mol, in reasonable agreement with the experimental overall rate for the catalytic cycle. For the other steps of the mechanism, only a preliminary investigation was made, indicating a few problems which require future QM/MM studies.