A Putative Amidase Endolysin Encoded by <i>Clostridium perfringens</i> St13 Exhibits Specific Lytic Activity and Synergizes with the Muramidase Endolysin Psm
<i>Clostridium perfringens</i> is an often-harmful intestinal bacterium that causes various diseases ranging from food poisoning to life-threatening fulminant disease. Potential treatments include phage-derived endolysins, a promising family of alternative antimicrobial agents. We survey...
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| Main Authors: | , , , , , |
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| Format: | Book |
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MDPI AG,
2021-03-01T00:00:00Z.
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| Summary: | <i>Clostridium perfringens</i> is an often-harmful intestinal bacterium that causes various diseases ranging from food poisoning to life-threatening fulminant disease. Potential treatments include phage-derived endolysins, a promising family of alternative antimicrobial agents. We surveyed the genome of the <i>C. perfringens</i> st13 strain and identified an endolysin gene, <i>psa</i>, in the phage remnant region. Psa has an N-terminal catalytic domain that is homologous to the amidase_2 domain, and a C-terminal domain of unknown function. <i>psa</i> and gene derivatives encoding various Psa subdomains were cloned and expressed in <i>Escherichia coli</i> as N-terminal histidine-tagged proteins. Purified His-tagged full-length Psa protein (Psa-his) showed <i>C. perfringens</i>-specific lytic activity in turbidity reduction assays. In addition, we demonstrated that the uncharacterized C-terminal domain has cell wall-binding activity. Furthermore, cell wall-binding measurements showed that Psa binding was highly specific to <i>C. perfringens</i>. These results indicated that Psa is an amidase endolysin that specifically lyses <i>C. perfringens</i>; the enzyme's specificity is highly dependent on the binding of the C-terminal domain. Moreover, Psa was shown to have a synergistic effect with another <i>C. perfringens</i>-specific endolysin, Psm, which is a muramidase that cleaves peptidoglycan at a site distinct from that targeted by Psa. The combination of Psa and Psm may be effective in the treatment and prevention of <i>C. perfringens</i> infections. |
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| Item Description: | 10.3390/antibiotics10030245 2079-6382 |