Effects of EGTA on cell surface structures of Corynebacterium glutamicum
The mycolic acid layer and S-layer of Corynebacterium glutamicum have been considered as permeability barriers against lytic agents. EGTA, a calcium chelator, inhibited C. glutamicum growth at relatively lower concentrations compared with other Gram-positive bacteria. We investigated the effect of EGTA on C. glutamicum cell surface structures. Simultaneous addition of EGTA and lysozyme resulted in cell lysis, whereas addition of these reagents separately had no such effect. Analysis of cell surface proteins showed that CspB, an S-layer protein, was released into the culture media and degraded to several sizes upon EGTA treatment. These findings suggest that EGTA treatment causes release and proteolysis of the CspB protein, resulting in increased cell surface permeability. FE-SEM visualization further confirmed alteration of cell surface structures in EGTA-treated cells. This is the first report suggesting the importance of calcium ions in cell surface integrity of C. glutamicum.
KeywordsCell wall Corynebacterium glutamicum CspB EGTA S-layer
This work was supported in part by a Grant-in-Aid for challenging Exploratory Research (25660088, to M. W.) from the Japan Society for Promotion of Science. The authors thank the members of the Suzukakedai Material Analysis Division, Technical Department, Tokyo Institute of Technology for FE-SEM analysis.
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