Induction of autolysis in Streptococcus faecium

Autolysis of exponential-phase Streptococcus faecium cells was promoted by pretreating the bacteria (freezing-thawing; -70 degrees C) in Tris buffer, followed by incubation at 37 degrees C in the same buffer. The effect was dependent on Tris concentration. The pretreatment provoked ultrastructurally visible damage with extensive loss of K+ and leakage of UV-absorbing components. No autolysis was observed when the bacteria frozen-thawed in Tris were incubated in the presence of the autolysin inhibitor N-bromosuccinimide nor when they had been grown in the presence of chloramphenicol or tetracycline. Furthermore, two autolytic-defective mutants, EC31 and EC78, isolated from S. faecium, did not autolyse when frozen-thawed and incubated in Tris. Freezing-thawing in Tris, however, imparted extensive cell damage to the mutants and to the antibiotic-treated bacteria as well as considerable leakage of K+ and UV-absorbing materials. These observations indicate that the lysis of S. faecium reported above is due to the activity of the endogenous bacterial autolysin. Induction of autolysis of S. faecium by freezing-thawing was also observed, although to a lesser extent, when Tris was replaced by imidazole.