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A novel toxin homologous to large clostridial cytotoxins found in culture supernatant of Clostridium perfringens type C

Kyoto Biken Laboratories, Inc., 24-16 Makishima-cho, Uji, Kyoto 611-0041, Japan

Correspondence
Katsuhiko Amimoto
por{at}kyotobiken.co.jp

An unknown cytotoxin was identified in the culture supernatant of Clostridium perfringens type C. The cytotoxin, named TpeL, which was purified using mAb-based affinity chromatography, had a lethal activity of 62 minimum lethal dose (MLD) mg^–1 in mice and a cytotoxic activity of 6.2x10⁵ cytotoxic units (CU) mg^–1 in Vero cells. The nucleotide sequence of TpeL was determined. The entire ORF had a length of 4953 bases, and the same nucleotide sequence was not recorded in the GenBank/EMBL/DDBJ databases. The molecular mass calculated from the deduced amino acid sequence was 191 kDa, and a signal peptide region was not found within the ORF. The deduced amino acid sequence exhibited 30–39 % homology to Clostridium difficile toxins A (TcdA) and B (TcdB), Clostridium sordellii lethal toxin (TcsL) and Clostridium novyi alpha-toxin (TcnA). The amino acid sequence of TpeL is shorter than these toxins, and the homologous region was located at the N-terminal site. Eighteen strains of C. perfringens types A, B and C were surveyed for the presence of the tpeL gene by PCR. The tpeL gene was detected in all type B (one strain) and C strains (five strains), but not in any type A strains (12 strains). TpeL was detected in culture filtrates of the five type C strains by dot-blot analysis, but not in the type B strain. It was concluded that TpeL is a novel toxin similar to the known large clostridial cytotoxins. Furthermore, the data indicated that TpeL is produced by many C. perfringens type C strains.

The GenBank/EMBL/DDBJ accession number for the tpeL sequence reported in this paper is AB262081.

Multiple sequence alignment data of this toxin and large clostridial cytotoxins are available as a supplementary figure with the online version of this paper.