A turbidimetric and electron microscopic study of the effects of several parameters on the lysis of Streptococcus faecalis by lysozyme

Abstract

The lytic effect of lysozyme on Streptococcus faecalis ATCC 9790 was studied by spectrophotometry and electron microscopy and it was found to be highly dependent on the ionic strength of the suspending media and on the ratio lysozyme to bacterial cell mass. When 7.2 X 10(8) bacteria/mL are exposed to 0.4 mg/mL of lysozyme in media with low ionic strength, the enzyme is bound in great amounts, as deduced from protein determinations and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS--PAGE); the binding prevents bacteriolysis in spite of the removal of the cell wall. Extensive lysis of S. faecalis could be obtained by reducing the ratio of lysozyme to bacterial cell mass. Stabilization of S. faecalis by lysozyme was also observed when exponential phase cells incubated under conditions that promote spontaneous autolysis (incubation in 0.05 M tris(hydroxymethyl)aminomethane buffer, pH 8.0, ionic strength = 0.01675) do not lyse and do not leak material which absorbs at 260 nm when lysozyme was present at the highest concentration.