title: Bacteria of the Roseobacter clade show potential for secondary metabolite production.
abstract: Members of the Roseobacter clade are abundant and widespread in marine habitats and have very diverse metabolisms. Production of acylated homoserine lactones (AHL) and secondary metabolites, e.g., antibiotics has been described sporadically. This prompted us to screen 22 strains of this group for production of signaling molecules, antagonistic activity against bacteria of different phylogenetic groups, and the presence of genes encoding for nonribosomal peptide synthetases (NRPS) and polyketide synthases (PKS), representing enzymes involved in the synthesis of various pharmaceutically important natural products. The screening approach for NRPS and PKS genes was based on polymerase chain reaction (PCR) with degenerate primers specific for conserved sequence motifs. Additionally, sequences from whole genome sequencing projects of organisms of the Roseobacter clade were considered. Obtained PCR products were cloned, sequenced, and compared with genes of known function. With the PCR approach genes showing similarity to known NRPS and PKS genes were found in seven and five strains, respectively, and three PKS and NRPS sequences from genome sequencing projects were obtained. Three strains exhibited antagonistic activity and also showed production of AHL. Overall production of AHL was found in 10 isolates. Phylogenetic analysis of the 16S rRNA gene sequences of the tested organisms showed that several of the AHL-positive strains clustered together. Three strains were positive for three or four categories tested, and were found to be closely related within the genus Phaeobacter. The presence of a highly similar hybrid PKS/NRPS gene locus of unknown function in sequenced genomes of the Roseobacter clade plus the significant similarity of gene fragments from the strains studied to these genes argues for the functional requirement of the encoded hybrid PKS/NRPS complex. Our screening results therefore suggest that the Roseobacter clade is indeed employing PKS/NRPS biochemistry and should thus be further studied as a potential and largely untapped source of secondary metabolites.
authors.affiliation: Institute for Chemistry and Biology of the Marine Environment (ICBM), University of Oldenburg, PO Box 2503, D-26111 Oldenburg, Germany.
authors.forename: Torben
authors.initials: T
authors.lastname: Martens
authors.forename: Lone
authors.initials: L
authors.lastname: Gram
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Grossart
authors.forename: Daniel
authors.initials: D
authors.lastname: Kessler
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
authors.forename: Meinhard
authors.initials: M
authors.lastname: Simon
authors.forename: Silke C
authors.initials: SC
authors.lastname: Wenzel
authors.forename: Thorsten
authors.initials: T
authors.lastname: Brinkhoff
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D010453:Peptide Synthases / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D048630:Polyketide Synthases / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D041904:Roseobacter / Q000145:classification / Q000201:enzymology / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
chemical_list: D048630:Polyketide Synthases; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase