title: A genomic screening approach to the structure-guided identification of drug candidates from natural sources.
abstract: The potential of actinomycetes to produce natural products has been exploited for decades. Recent genomic sequence analyses have revealed a previously unrecognized biosynthetic potential and diversity. In order to rationally exploit this potential, we have developed a sequence-guided genetic screening strategy. In this "genome mining" approach, genes that encode tailoring enzymes from natural product biosyntheses pathways serve as indicator genes for the identification of strains that have the genetic potential to produce natural products of interest. We chose halogenases, which are known to be involved in the synthesis of halometabolites as representative examples. From PCR screening of 550 randomly selected actinomycetes strains, we identified 103 novel putative halogenase genes. A phylogenetic analysis of the corresponding putative halogenases, and the determination of their sequential context with mass spectrometric analysis of cultures filtrates revealed a distinct correlation between the sequence and secondary metabolite class of the halometabolite. The described screening strategy allows rapid access to novel natural products with predetermined structural properties.
authors.affiliation: Combinature Biopharm AG, Robert-Roessle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hornung
authors.forename: Marcelo
authors.initials: M
authors.lastname: Bertazzo
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Dziarnowski
authors.forename: Kathrin
authors.initials: K
authors.lastname: Schneider
authors.forename: Katrin
authors.initials: K
authors.lastname: Welzel
authors.forename: Sven-Eric
authors.initials: SE
authors.lastname: Wohlert
authors.forename: Meike
authors.initials: M
authors.lastname: Holzenkämpfer
authors.forename: Graeme J
authors.initials: GJ
authors.lastname: Nicholson
authors.forename: Andreas
authors.initials: A
authors.lastname: Bechthold
authors.forename: Roderich D
authors.initials: RD
authors.lastname: Süssmuth
authors.forename: Andreas
authors.initials: A
authors.lastname: Vente
authors.forename: Stefan
authors.initials: S
authors.lastname: Pelzer
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001688:Biological Products; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D023281:Genomics; D009682:Magnetic Resonance Spectroscopy; D008403:Mass Screening; D008969:Molecular Sequence Data; D015394:Molecular Structure; D004364:Pharmaceutical Preparations; D017386:Sequence Homology, Amino Acid
chemical_list: D001688:Biological Products; D017931:DNA Primers; D004364:Pharmaceutical Preparations