Lactoperoxidase-catalyzed oxidation of thiocyanate: polarographic study of the oxidation products.

The lactoperoxidase-catalyzed oxidation of thiocyanate (SCN-) was studied by two different polarographic techniques: direct current polarography and linear sweep voltammetry. The main oxidation product at pH 6.5, with a half-wave potential (E1/2) of -0.39 to -0.44 V, was identified as hypothiocyanite (OSCN-) ion. The E1/2 for OSCN- was not available in the literature. The identification of OSCN- was based on a close correlation between the current of the OSCN- peak and the concentration of chemically assayed OSCN-. Also the specific rates of decay of the current and that of chemically detectable OSCN- were similar, and both curves followed apparent first-order kinetics. Subsequently, the addition of a reducing agent (2-mercaptoethanol) resulted in immediate disappearance of both chemically detectable OSCN- and the OSCN- wave in the polarograms. All three components of the lactoperoxidase (LPO) system (SCN-, H2O2, and LPO) were needed to produce the OSCN- peak. Addition of excess H2O2 or H2O2-LPO to an OSCN--SCN- mixture resulted in a formation of a new peak with a characteristic peak potential (Ep) of -0.20 to -0.25 V. The generation of this new peak was associated with a simultaneous, markedly enhanced decrease of OSCN- concentration, indicating a possible reaction between H2O2 (or H2O2-LPO) and OSCN-. No equivalent reaction was obtained by the addition of buffer alone. This new peak may represent higher oxy acids of SCN- (O2SCN-, O3SCN-), formed in the oxidation of OSCN- by H2O2 or by H2O2-LPO. This type of reaction can explain why, in solutions which already contain OSCN- (e.g., in saliva), the addition of H2O2 results in the formation of highly reactive, short-lived antimicrobial products in addition to OSCN-.