Applied Microbiology and Biotechnology

, Volume 90, Issue 6, pp 1973–1979

Identification and characterization of a putative endolysin encoded by episomal phage phiSM101 of Clostridium perfringens

  • Hirofumi Nariya
  • Shigeru Miyata
  • Eiji Tamai
  • Hiroshi Sekiya
  • Jun Maki
  • Akinobu Okabe
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-011-3253-z

Cite this article as:
Nariya, H., Miyata, S., Tamai, E. et al. Appl Microbiol Biotechnol (2011) 90: 1973. doi:10.1007/s00253-011-3253-z

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.

Keywords

Clostridium perfringensBacteriophageEndolysinN-acetylmuramidaseN-acetylmuramoyl-l-alanine amidase

Supplementary material

253_2011_3253_MOESM1_ESM.pdf (364 kb)
ESM 1(PDF 364 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Hirofumi Nariya
    • 1
  • Shigeru Miyata
    • 1
  • Eiji Tamai
    • 2
  • Hiroshi Sekiya
    • 2
  • Jun Maki
    • 2
  • Akinobu Okabe
    • 1
  1. 1.Department of Microbiology, Faculty of MedicineKagawa UniversityKita-gunJapan
  2. 2.Department of Infectious Disease, College of Pharmaceutical SciencesMatsuyama UniversityMatsuyamaJapan