Document Detail


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.
MedLine Citation:
PMID:  20038713     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cereulide, the emetic Bacillus 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 the ces gene locus encoding cereulide synthetase; however, detailed information about the transcriptional organization of the ces genes was lacking. The present study shows that the cesPTABCD genes are transcribed as a 23-kb polycistronic transcript, while cesH, 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 the cesP coding sequence, encoding a 4'-phosphopantetheinyl transferase. This promoter drives transcription of cesPTABCD. In addition, intracistronic promoter regions in proximity to the translational start sites of cesB and cesT were identified but were only weakly active under the chosen assay conditions. The identified main promoter was amplified from the emetic reference strain B. 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 of ces gene expression in complex matrices.
Authors:
Monica K Dommel; Elrike Frenzel; Bernd Strasser; Claudia Blöchinger; Siegfried Scherer; Monika Ehling-Schulz
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-12-28
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  76     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2010 Feb 
Date Detail:
Created Date:  2010-02-08     Completed Date:  2010-04-02     Revised Date:  2010-09-27    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1232-40     Citation Subset:  IM    
Affiliation:
Microbiology Unit, Center for Nutrition and Food Research ZIEL, Technische Universität München, 85350 Freising, Germany.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Bacillus cereus / genetics*,  growth & development,  metabolism,  pathogenicity*
Bacterial Toxins / biosynthesis*,  toxicity
Base Sequence
DNA Primers / genetics
DNA, Bacterial / genetics
Depsipeptides / biosynthesis*,  toxicity
Environmental Monitoring / methods
Food Microbiology*
Foodborne Diseases / etiology
Gene Expression
Genes, Bacterial
Genes, Reporter
Humans
Molecular Sequence Data
Multigene Family
Peptide Synthases / genetics,  metabolism
Promoter Regions, Genetic
Vomiting / etiology
Chemical
Reg. No./Substance:
0/Bacterial Toxins; 0/DNA Primers; 0/DNA, Bacterial; 0/Depsipeptides; 0/cereulide; EC 6.3.2.-/Peptide Synthases; EC 6.3.2.-/non-ribosomal peptide synthase
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