Heme models of peroxidase enzymes: deuteroferriheme-catalyzed chlorination of monochlorodimedone by sodium chlorite

The iron(III) complex of deuteroporphyrin(IX), deuteroferriheme, catalyzes the chlorination, by sodium chlorite, of the active methylene compound monochlorodimedone (MCD) to dichlorodimedone. Rate studies, carried out on a stopped-flow spectrophotometric time scale, show the chlorination to be zero-order in MCD, first-order in ClO2- and to display a complex dependence on heme. The active chlorinating agent is believed to be hypochlorite, OCl-, formed as a result of the initial two-electron oxidation of heme to peroxidatic intermediate by chlorite ion. This scheme is supported by the fact that the normal (4:1) heme:ClO2- molar stoichiometry is reduced in the presence of MCD to values approaching 2:1. This suggests that MCD is an effective scavenger of OCl-, which, in the absence of active methylene compound, serves as a two-electron oxidant of heme. The zero-order dependence of rate on MCD is attributed to the slow formation of OCl-, consequent to a mechanism in which the rate-limiting step is viewed to be the regeneration of free heme from peroxidatic intermediate, probably via a catalatic pathway. Support for such a mechanism is provided by the fact that addition of ascorbate greatly enhances the rate of MCD chlorination, presumably by accelerating the rate of heme regeneration via perioxidation reduction of the heme intermediate.