Lactoperoxidase, peroxide, thiocyanate antimicrobial system: correlation of sulfhydryl oxidation with antimicrobial action

Abstract

The antimicrobial activity of the lactoperoxidase, peroxide, thiocyanate system against Escherichia coli was directly related to the oxidation of bacterial sulfhydryls. Lactoperoxidase catalyzed the oxidation of thiocyanate, which resulted in the accumulation of hypothiocyanite ion, OSCN-. A portion of the bacterial sulfhydryls were oxidized by OSCN- to yield sulfenic acid and sulfenyl thiocyanate derivatives. The remaining sulfhydryls were not oxidized, although OSCN- was present in large excess. The oxidation of sulfhydryls to sulfenyl derivatives inhibited bacterial respiration. This inhibition could be reversed by adding sulfhydryl compounds to reduce the sulfenyl derivatives and the excess OSCN-. Also, this inhibition could be reversed by washing the cells so as to remove the excess unreacted OSCN-. After washing, the bacteria underwent a time-dependent recovery of their sulfhydryl content. This recovery resulted in recovery of the ability to respire. The inhibited cells were viable if diluted and plated shortly after the incubation with the lactoperoxidase, peroxide, thiocyanate system. On the other hand, long-term incubation in the presence of the excess OSCN- resulted in loss of viability. Also, the inhibition of respiration became irreversible. During this long-term incubation, the excess OSCN- was consumed and the sulfenyl derivatives disappeared.