title: Functional and structural characterization of soluble recombinant epsilon toxin of Clostridium perfringens D, causative agent of enterotoxaemia.
abstract: Clostridium perfringens types B and D are responsible for enterotoxaemia, one of the major causes of cattle mortality and is therefore of great economic concern. The epsilon toxin produced by the organism is the major antigenic determinant and has been directly implicated for the disease causation. In the present paper, we evaluated the biological activity of the recombinant epsilon toxin (rEtx) produced as soluble protein in Escherichia coli. The rEtx was purified to near homogeneity by a one-step anion-exchange chromatography. The immunological identity of purified rEtx was confirmed by Western blotting using a monoclonal antibody against the native toxin. The rEtx formed heptamer in the Madin-Darby canine kidney (MDCK) cells and synaptosomal membrane of mouse brain and was cytotoxic to the MDCK cells with a CT(50) of 30 ng/ml. The rEtx was highly stable and its thermostability profile related well with its biological activity. The rEtx was purified in large amounts and exhibited all the properties of native toxin and therefore can be used for the development of vaccine against the pathogen.
authors.affiliation: Gene Regulation Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Deepika Dayal
authors.initials: DD
authors.lastname: Mathur
authors.forename: Sachin
authors.initials: S
authors.lastname: Deshmukh
authors.forename: Himani
authors.initials: H
authors.lastname: Kaushik
authors.forename: Lalit C
authors.initials: LC
authors.lastname: Garg
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003016:Clostridium perfringens / Q000378:metabolism*; D004285:Dogs; D004926:Escherichia coli / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; C039811:Clostridium perfringens epsilon-toxin; D011994:Recombinant Proteins