title: Structural and functional organization of the fengycin synthetase multienzyme system from Bacillus subtilis b213 and A1/3.
abstract: BACKGROUND
  Bacillus subtilis strains produce a broad spectrum of lipopeptides that are potent biosurfactants and have specific antimicrobial and antiviral activities. The cyclic lipodecapeptide fengycin is one such compound. Although the fengycin biosynthetic genes in B. subtilis 168 (pps genes) and F29-3 (fen genes) have been well characterized, only limited information is available about the biochemical features of the fengycin synthetase multienzyme system.
  
  
  RESULTS
  Five multifunctional peptide synthetases (Fen1-5) that catalyze biosynthesis of the peptide portion of fengycin have been purified from crude extracts of the B. subtilis b213 and A1/3 strains. These enzymes activate all fengycin amino-acid components as aminoacyl adenylates or aminoacyl thioesters. Fen1, Fen2 and Fen3 are each approximately 286 kDa, Fen4 is approximately 400 kDa and Fen 5 is approximately 140kDa; each enzyme activates a different set of L-amino acids. A five-gene cluster (fen1-5) was detected in the B. subtilis A1/3 genome that shows high homology to the pps and fen genes in B. subtilis strains 168 and F29-3. Disruption of fen4 resulted in a loss of fengycin production. The fengycin synthetase enzymes isolated from B. subtilis b213 were assigned to the corresponding A1/3 fen genes by their amino-terminal sequences.
  
  
  CONCLUSIONS
  The structural and functional organization of the fengycin synthetase system from B. subtilis b213 has been characterized in detail and correlated with the corresponding pps and fen genes in B. subtilis strains 168, A1/3 and F29-3. Biosynthesis of the peptide part of fengycin involves five multifunctional modular proteins that assemble the lipopeptide chain using a nonribosomal, multiple carrier thiotemplate mechanism.
authors.affiliation: Max-Volmer-Institut für Biophysikalische Chemie und Biochemie, Fachgebiet Biochemie und Molekulare Biologie, Technische Universität Berlin, Franklinstrasse 29, D-10587 Berlin, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Steller
authors.forename: D
authors.initials: D
authors.lastname: Vollenbroich
authors.forename: F
authors.initials: F
authors.lastname: Leenders
authors.forename: T
authors.initials: T
authors.lastname: Stein
authors.forename: B
authors.initials: B
authors.lastname: Conrad
authors.forename: J
authors.initials: J
authors.lastname: Hofemeister
authors.forename: P
authors.initials: P
authors.lastname: Jacques
authors.forename: P
authors.initials: P
authors.lastname: Thonart
authors.forename: J
authors.initials: J
authors.lastname: Vater
mesh_terms: D000595:Amino Acid Sequence; D000903:Antibiotics, Antineoplastic / Q000096:biosynthesis; D000935:Antifungal Agents / Q000096:biosynthesis; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis; D055666:Lipopeptides; D008074:Lipoproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D005810:Multigene Family; D009154:Mutation; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D010456:Peptides, Cyclic; D017386:Sequence Homology, Amino Acid
chemical_list: D000903:Antibiotics, Antineoplastic; D000935:Antifungal Agents; D001426:Bacterial Proteins; D055666:Lipopeptides; D008074:Lipoproteins; D009097:Multienzyme Complexes; D010456:Peptides, Cyclic; C049972:fengycin; C009365:surfactin peptide; D010453:Peptide Synthases; C110972:fengycin synthetase