Journal
TOXINS
Volume 13, Issue 10, Pages -Publisher
MDPI
DOI: 10.3390/toxins13100698
Keywords
Bacillus cereus; Bacillus cytotoxicus; CytK-1; cytotoxicity; Caco-2
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Funding
- International Office for Cooperation of Universite Catholique de Louvain, Belgium (UCLouvain)
- National Foundation for Scientific Research (FNRS)
- Special Research Fund (Ghent University) [BOFSTA2017004201]
- European Unions Horizon 2020 research and innovation programme project FoodEnTwin [810752]
- GlobalMinds project of Ghent University
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The study demonstrated that the cytotoxicity of Bacillus cytotoxicus is primarily attributed to CytK-1, with knockout of this gene significantly reducing toxicity and complementation restoring cytotoxicity.
The thermotolerant representative of the Bacillus cereus group, Bacillus cytotoxicus, reliably harbors the coding gene of cytotoxin K-1 (CytK-1). This protein is a highly cytotoxic variant of CytK toxin, initially recovered from a diarrheal foodborne outbreak that caused the death of three people. In recent years, the cytotoxicity of B. cytotoxicus has become controversial, with some strains displaying a high cytotoxicity while others show no cytotoxicity towards cell lines. In order to better circumscribe the potential pathogenic role of CytK-1, knockout (KO) mutants were constructed in two B. cytotoxicus strains, E8.1 and E28.3. The complementation of the cytK-1 KO mutation was implemented in a mutant strain lacking in the cytK-1 gene. Using the tetrazolium salt (MTT) method, cytotoxicity tests of the cytK-1 KO and complemented mutants, as well as those of their wild-type strains, were carried out on Caco-2 cells. The results showed that cytK-1 KO mutants were significantly less cytotoxic than the parental wild-type strains. However, the complemented mutant was as cytotoxic as the wild-type, suggesting that CytK-1 is the major cytotoxicity factor in B. cytotoxicus.
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