Inhibition of air-exposure stress–Induced autolysis in Clostridium perfringens by Zn2+

Nozomu Matsunaga, Seira Egusa, Riyo Aono, Eiji Tamai, Yasuo Hitsumoto, and Seiichi Katayama

Clostridium perfringens is a pathogenic anaerobe that causes gas gangrene and food poisoning. Although autolysin-
mediated reorganization of the bacterial cell wall is crucial for cell division, excessive autolysin activity
induced by stressors can lead to cell lysis. In C. perfringens, air exposure is a significant stressor that causes cell
lysis, and Acp (N-acetylglucosaminidase) is known to be a major autolysin. To further facilitate C. perfringens
research, a technology to prevent air-induced cell lysis must be developed. This study investigated the role of
Acp in air-induced autolysis and explored potential inhibitors that would prevent cell lysis during experimental
procedures. Morphological analyses confirmed that Acp functions as an autolysin in C. perfringens, as acpdeficient
strains exhibited filamentous growth. The mutants exhibited negligible autolysis under air-exposure
stress, confirming the involvement of Acp in the autolytic process. We also evaluated the effects of various divalent
cations on Acp activity in vitro and identified Zn2+ as a potent inhibitor. Brief treatment with a Zn2+-
containing buffer induced dose-dependent cell elongation and autolysis inhibition in C. perfringens. These
findings demonstrate that simple Zn2+ treatment before experiments stabilizes C. perfringens cells, reducing
autolysis under aerobic conditions and facilitating various biological studies, except morphological analyses.

Acta Medica Okayama

Okayama University Medical School

79

5

345

352

0386-300X

10.18926/AMO/69435

41126465