RRC ID 17436
Author Nariya H, Miyata S, Tamai E, Sekiya H, Maki J, Okabe A.
Title Identification and characterization of a putative endolysin encoded by episomal phage phiSM101 of Clostridium perfringens.
Journal Appl Microbiol Biotechnol
Abstract Clostridium perfringens produces potent toxins and histolytic enzymes, causing various diseases including life-threatening fulminant diseases in humans and other animals. Aiming at utilizing a phage endolysin as a therapeutic alternative to antibiotics, we surveyed the genome and bacteriophage sequences of C. perfringens. A phiSM101 muramidase gene (psm) revealed by this study can be assumed to encode an N-acetylmuramidase, since the N-terminal catalytic domain deduced from the gene shows high homology of those of N-acetylmuramidases. The psm gene is characteristic in that it is present in phiSM101, an episomal phage of enterotoxigenic C. perfringens type A strain, SM101, and also in that homologous genes are present in the genomes of all five C. perfringens toxin types. The psm gene was cloned and expressed in Escherichia coli as a protein histidine-tagged at the N-terminus (Psm-his). Psm-his was purified to homogeneity by nickel-charged immobilized metal affinity chromatography and anion-exchange chromatography. The purified enzyme lysed cells of all C. perfringens toxin types but not other clostridial species tested, as was shown by a turbidity reduction assay. These results indicate the Psm-his is useful as a cell-wall lytic enzyme and also suggest that it is potentially useful for biocontrol of this organism.
Volume 90(6)
Pages 1973-9
Published 2011-6-1
DOI 10.1007/s00253-011-3253-z
PMID 21484204
MeSH Bacteriolysis* Bacteriophages / enzymology* Bacteriophages / isolation & purification Chromatography, Affinity / methods Cloning, Molecular Clostridium perfringens / virology* Endopeptidases / genetics Endopeptidases / isolation & purification Endopeptidases / metabolism* Escherichia coli / genetics Gene Expression Glycoside Hydrolases / genetics Glycoside Hydrolases / isolation & purification Glycoside Hydrolases / metabolism* Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism
IF 3.53
Times Cited 24
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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General Microbes JCM 1296 JCM 1403 JCM 1419