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Cereulide, the emeticBacillus cereus toxin, is synthesized by cereulide synthetase via a nonribosomal peptide synthetase (NRPS) mechanism. Previous studies focused on the identification, structural organization, and biochemical characterization of theces gene locus encoding cereulide synthetase; however, detailed information about the transcriptional organization of theces genes was lacking. The present study shows that thecesPTABCD genes are transcribed as a 23-kb polycistronic transcript, whilecesH , encoding a putative hydrolase, is transcribed from its own promoter. Transcription initiation was mapped by primer extension and rapid amplification of cDNA ends (RACE). Deletion analysis of promoter elements revealed a main promoter located upstream of thecesP coding sequence, encoding a 4′-phosphopantetheinyl transferase. This promoter drives transcription ofcesPTABCD . In addition, intracistronic promoter regions in proximity to the translational start sites ofcesB andcesT were identified but were only weakly active under the chosen assay conditions. The identified main promoter was amplified from the emetic reference strainB. cereus F4810/72 and fused to luciferase genes in order to study promoter activity in complex environments and to establish a biomonitoring system to assess cereulide production in different types of foods.ces promoter activity was strongly influenced by the food matrix and varied by 5 orders of magnitude. The amount of cereulide toxin extracted from spiked foods correlated well with the bioluminescence data, thus illustrating the potential of the established reporter system for monitoring ofces gene expression in complex matrices.

applied and environmental microbiology

Identification of the Main Promoter Directing Cereulide Biosynthesis in Emetic <i>Bacillus cereus</i> and Its Application for Real-Time Monitoring of <i>ces</i> Gene Expression in Foods

Identification of the Main Promoter Directing Cereulide Biosynthesis in Emetic Bacillus cereus and Its Application for Real-Time Monitoring of ces Gene Expression in Foods