title: Cereulide synthetase gene cluster from emetic Bacillus cereus: structure and location on a mega virulence plasmid related to Bacillus anthracis toxin plasmid pXO1.
abstract: BACKGROUND
  Cereulide, a depsipeptide structurally related to valinomycin, is responsible for the emetic type of gastrointestinal disease caused by Bacillus cereus. Recently, it has been shown that this toxin is produced by a nonribosomal peptide synthetase (NRPS), but its exact genetic organization and biochemical synthesis is unknown.
  
  
  RESULTS
  The complete sequence of the cereulide synthetase (ces) gene cluster, which encodes the enzymatic machinery required for the biosynthesis of cereulide, was dissected. The 24 kb ces gene cluster comprises 7 CDSs and includes, besides the typical NRPS genes like a phosphopantetheinyl transferase and two CDSs encoding enzyme modules for the activation and incorporation of monomers in the growing peptide chain, a CDS encoding a putative hydrolase in the upstream region and an ABC transporter in the downstream part. The enzyme modules responsible for incorporation of the hydroxyl acids showed an unusual structure while the modules responsible for the activation of the amino acids Ala and Val showed the typical domain organization of NRPS. The ces gene locus is flanked by genetic regions with high homology to virulence plasmids of B. cereus, Bacillus thuringiensis and Bacillus anthracis. PFGE and Southern hybridization showed that the ces genes are restricted to emetic B. cereus and indeed located on a 208 kb megaplasmid, which has high similarities to pXO1-like plasmids.
  
  
  CONCLUSIONS
  The ces gene cluster that is located on a pXO1-like virulence plasmid represents, beside the insecticidal and the anthrax toxins, a third type of B. cereus group toxins encoded on megaplasmids. The ces genes are restricted to emetic toxin producers, but pXO1-like plasmids are also present in emetic-like strains. These data might indicate the presence of an ancient plasmid in B. cereus which has acquired different virulence genes over time. Due to the unusual structure of the hydroxyl acid incorporating enzyme modules of Ces, substantial biochemical efforts will be required to dissect the complete biochemical pathway of cereulide synthesis.
authors.affiliation: Lehrstuhl für Mikrobielle Okologie, Department für Biowissenschaftliche Grundlagen, WZW, Technische Universität München, Freising, Germany. Monika.Ehling-Schulz@wzw.tum.de
authors.forename: Monika
authors.initials: M
authors.lastname: Ehling-Schulz
authors.forename: Martina
authors.initials: M
authors.lastname: Fricker
authors.forename: Harald
authors.initials: H
authors.lastname: Grallert
authors.forename: Petra
authors.initials: P
authors.lastname: Rieck
authors.forename: Martin
authors.initials: M
authors.lastname: Wagner
authors.forename: Siegfried
authors.initials: S
authors.lastname: Scherer
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics*; D001408:Bacillus anthracis / Q000235:genetics* / Q000472:pathogenicity; D001409:Bacillus cereus / Q000235:genetics* / Q000472:pathogenicity; D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D047630:Depsipeptides / Q000096:biosynthesis*; D016521:Electrophoresis, Gel, Pulsed-Field; D008969:Molecular Sequence Data; D005810:Multigene Family; D009693:Nucleic Acid Hybridization; D009876:Operon; D010453:Peptide Synthases / Q000235:genetics*; D010957:Plasmids; D017434:Protein Structure, Tertiary; D014774:Virulence
chemical_list: D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D047630:Depsipeptides; C030325:anthrax toxin; C089294:cereulide; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase