[ODP-107] Involvement of autolysin in cell lysis of Clostridium perfringens cells
Clostridium perfringens is an anaerobic bacterium which lyses by the oxygen stress or other factors including Triton X-100. Autolysin (Acp) is thought to be involved in these cell lyses. We investigated the involvement of Acp in both spontaneous bacteriolysis and Triton-induced lysis of C. perfringens cells and the effect of Zn on these lyses. Wild type C. perfringens 13 and Acp-deficient mutant C. perfringens 13 acp::erm cells were cultured in GAM medium up to OD600 = 1.0 ± 0.1. Cell suspension in a microplate was incubated at 37 degree Celsius in the presence or absence of 0.05% Triton X-100 and then the absorbance at 600 nm was measured. In some experiments, cell lysis assay was performed in the presence of various concentrations of ZnCl2. Wild type C. perfringens 13 cells lysed spontaneously (40-60% lysis occurred by 60 min incubation) whereas C. perfringens 13 acp::erm cells did not. This result suggested the involvement of Acp in the oxygen induced cell lysis. The spontaneous cell lysis of C. perfringens 13 was inhibited in the presence of more than 2 mM ZnCl2, indicating that the enzymatic activity of Acp was inhibited by Zn. Because not only C. perfringens 13 celell lysisls but also C. perfringens 13 acp::erm cells lysed by the presence of Triton, Acp may not be involved in the Triton-induced bacteriolysis.