Phospholipase C produced by clostridium botulinum types C and D: Comparison of gene, enzymatic, and biological activities with those of Clostridium perfringens alpha-toxin

Ni Nengah Dwi Fatmawati, Yoshihiko Sakaguchi, Tomonori Suzuki, Masataka Oda, Kenta Shimizu, Yumiko Yamamoto, Jun Sakurai, Osamu Matsushita, Keiji Oguma

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Clostridium botulinum type C and D strains recently have been found to produce PLC on egg yolk agar plates. To characterize the gene, enzymatic and biological activities of C. botulinum PLCs (Cb-PLCs), the cb-plc genes from 8 strains were sequenced, and 1 representative gene was cloned and expressed as a recombinant protein. The enzymatic and hemolytic activities of the recombinant Cb-PLC were measured and compared with those of the Clostridium perfringens alpha-toxin Each of the eight cb-plc genes encoded a 399 amino acid residue protein preceded by a 27 residue signal peptide. The protein consists of 2 domains, the N- and C-domains, and the overall amino acid sequence identity between Cb-PLC and alpha-toxin was greater than 50%, suggesting that Cb-PLC is homologous to the alpha-toxin. The key residues in the N-domain were conserved, whereas those in the C-domain which are important in membrane interaction were different than in the alpha-toxin. As expected, Cb-PLC could hydrolyze egg yolk phospholipid, p-nitrophenylphosphorylcholine, and sphingomyelin, and also exhibited hemolytic activity; however, its activities were about 4- to over 200-fold lower than those of alpha-toxin. Although Cb-PLC showed weak enzymatic and biological activities, it is speculated that Cb-PLC might play a role in the pathogenicity of botulism or for bacterial survival.

Original languageEnglish
Pages (from-to)9-18
Number of pages10
JournalActa medica Okayama
Volume67
Issue number1
Publication statusPublished - Apr 10 2013

Keywords

  • Botulinum phospholipase C
  • Botulinum toxin
  • Hemolytic activity
  • Phospholipase C activity
  • Sphingomyelinase activity

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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