Inhibition of Streptococcus mutans by the lactoperoxidase antimicrobial system

Abstract

Inhibition of bacterial metabolism by the lactoperoxidase (LP)-hydrogen peroxide (H2O2)-thiocyanate system was studied with representatives of serotypes a through g of Streptococcus mutans. The aims were to determine whether the amount of H2O2 released from these catalase-negative bacteria is sufficient to activate the LP system and whether these oral bacteria are resistant to inhibition by the LP system, which is active in human saliva. When the washed, stationary-phase cells were incubated aerobically with LP, thiocyanate, and glucose (Glc), greater than 90% inhibition of Glc utilization and lactate production was obtained with strains that released large amounts of H2O2 (BHT, FA-1, OMZ-176); 20 to 50% inhibition was obtained with strains that released about half as much H2O2 (B-13, Ingbritt); and no inhibition was obtained with strains that released only small amounts of H2O2 (AHT, HS-6, GS-5, LM-7, OMZ-175, 6715-15). Inhibition was most effective at pH 5, whereas release of H2O2 and accumulation of the inhibitor (hypothiocyanite ion) were highest at pH 8. With H2O2-releasing cells from early stationary phase, preincubation with Glc abolished inhibition, though it did not influence H2O2 release. Cells harvested 24 h later were depleted of sulfhydryl compounds. Inhibition of these cells was abolished by preincubation with Glc and certain sulfhydryl or disulfide compounds (reduced or oxidized glutathione, cysteine or cystine). This preincubation increased cell sulfhydryl content but had no effect on H2O2 release. All strains were inhibited when incubated with LP, thiocyanate, and added (exogenous) H2O2. Smaller amounts of H2O2 were required to inhibit at pH 5, and larger amounts were required to inhibit cells preincubated with Glc or with Glc and the sulfhydryl or disulfide compounds. The results indicate that pH, amount of H2O2, cell sulfhydryl content, and stored-carbohydrate content determine susceptibility to inhibition.