Bacillus cereus Toxin Repertoire: Diversity of (Iso)cereulide(s)

The emetic Bacillus cereus toxin cereulide (1) poses a significant safety risk in the food industry, causing emesis and nausea after consumption of contaminated foods. Analogously to cereulide, the structures of various isocereulides, namely, isocereulides A–G, have been recently reported and could also be identified in B. cereus-contaminated food samples. The HPLC fractiona-tion of B. cereus extracts allows us to isolate additional isocereulides. By applying MSⁿ sequencing, post-hydrolytic dipeptide, amino acid and α-hydroxy acid analyses using UPLC-ESI-TOF-MS to purify the analytes, seven new isocereulides H–N (2–8) could be elucidated in their chemical struc-tures. The structure elucidation was supported by one-dimensional and two-dimensional NMR spectra of the isocereulides H (2), K (5), L and N (6 + 8) and M (7). The toxicity of 2–8 was investigated in a HEp-2 cell assay to determine their respective 50% effective concentration (EC50). Thus, 2–8 exhibited EC50 values ranging from a 0.4-to 1.4-fold value compared to cereulide (1). Missing struc-ture-activity correlations indicate the necessity to determine the toxic potential of all naturally pre-sent isocereulides as single compounds to be able to perform a thorough toxicity evaluation of B. cereus-contaminated foods in the future.