Synergism of lysozyme, proteases and inorganic monovalent anions in the bacteriolysis of oral Streptococcus mutans GS5.

Streptococcus mutans GS5 was grown in synthetic medium containing radioactive thymidine to monitor deoxyribonucleic acid release. At neutral pH, cell lysis of hen egg-white lysozyme- or lysozyme-protease-treated cells was dependent upon the nature and concentration of the additive inorganic anions, HCO-3, SCN-, Cl- or F-. At acidic pH, NaHCO3, but not NaSCN, NaCl or NaF, was effective in promoting cell lysis which was due not only to the change in pH but also to the new HCO-3 anion concentration at the new pH. In both pH 4 and 5.2 reaction mixtures, the lysozyme and trypsin acted synergistically with NaHCO3 and the amount of lysis produced was markedly greater than in reaction mixtures containing lysozyme and bicarbonate but no protease. At apparent sub-lytic concentrations of NaHCO3, lysis was achieved by adding an appropriate concentration of one of NaSCN, NaCl or NaF to the lysozyme-protease-damaged cells. Thiocyanate proved to be most effective among the anions requiring lower concentrations to elicit lysis compared to chloride or fluoride for a fixed sub-lytic concentration of bicarbonate. As the NaHCO3 concentration increased, the lysis in the presence of these other anions increased until maximum levels of released deoxyribonucleic acid (DNA) were attained. In addition, the higher the NaHCO3 concentration, the more marked was the change in the degree of cell lysis. At a selected concentration at which NaHCO3 was not effective with any one salt, lysis could be achieved by combining all four inorganic anions at this concentration. The results suggest that the various anions present in oral fluids may together be sufficient to trigger lysis of oral microorganisms.