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The major virulence factors of <i>Clostridioides difficile</i> (<i>C. difficile</i>) are enterotoxins A (TcdA) and B (TcdB). The study of toxins is a crucial step in exploring the virulence of this pathogen. Currently, the toxin purification process is either laborious and time-consuming in <i>C. difficile</i> or performed in heterologous hosts. Therefore, we propose a streamlined method to obtain functional toxins in <i>C. difficile</i>. Two <i>C. difficile</i> strains were generated, each harboring a sequence encoding a His-tag at the 3′ end of <i>C. difficile</i> 630∆<i>erm tcdA</i> or <i>tcdB</i> genes. Each toxin gene is expressed using the P_tet promoter, which is inducible by anhydro-tetracycline. The obtained purification yields were 0.28 mg and 0.1 mg per liter for rTcdA and rTcdB, respectively. In this study, we successfully developed a simple routine method that allows the production and purification of biologically active rTcdA and rTcdB toxins with similar activities compared to native toxins.