title: Development of a Biosensor Concept to Detect the Production of Cluster-Specific Secondary Metabolites.
abstract: Genome mining of actinomycete bacteria aims at the discovery of novel bioactive secondary metabolites that can be developed into drugs. A new repressor-based biosensor to detect activated secondary metabolite biosynthesis gene clusters in Streptomyces was developed. Biosynthetic gene clusters for undecylprodigiosin and coelimycin in the genome of Streptomyces lividans TK24, which encoded TetR-like repressors and appeared to be almost "silent" based on the RNA-seq data, were chosen for the proof-of-principle studies. The bpsA reporter gene for indigoidine synthetase was placed under control of the promotor/operator regions presumed to be controlled by the cluster-associated TetR-like repressors. While the biosensor for undecylprodigiosin turned out to be nonfunctional, the coelimycin biosensor was shown to perform as expected, turning on biosynthesis of indigoidine in response to the concomitant production of coelimycin. The developed reporter system concept can be applied to those cryptic gene clusters that encode metabolite-sensing repressors to speed up discovery of novel bioactive compounds in Streptomyces.
authors.affiliation: Department of Biotechnology, Norwegian University of Science and Technology , 7491 Trondheim, Norway.|The Department of Laboratory Medicine, Children's and Women's Health (LBK), Norwegian University of Science and Technology , 7491 Trondheim, Norway.
authors.forename: Yi-Qian
authors.initials: YQ
authors.lastname: Sun
authors.affiliation: Center for Biotechnology, Bielefeld University , Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Busche
authors.affiliation: Center for Biotechnology, Bielefeld University , Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Rückert
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland , Actinobacteria Metabolic Engineering Group, 66123 Saarbrücken, Germany.|Universität des Saarlandes , Pharmaceutical Biotechnology, 66123 Saarbrücken, Germany.
authors.forename: Constanze
authors.initials: C
authors.lastname: Paulus
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland , Actinobacteria Metabolic Engineering Group, 66123 Saarbrücken, Germany.
authors.forename: Yuriy
authors.initials: Y
authors.lastname: Rebets
authors.affiliation: Institute of Molecular Biology, Slovak Academy of Sciences , 845 51 Bratislava, Slovak Republic.
authors.forename: Renata
authors.initials: R
authors.lastname: Novakova
authors.affiliation: Center for Biotechnology, Bielefeld University , Universitätsstraße 27, 33615 Bielefeld, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Kalinowski
authors.affiliation: Helmholtz Institute for Pharmaceutical Research Saarland , Actinobacteria Metabolic Engineering Group, 66123 Saarbrücken, Germany.|Universität des Saarlandes , Pharmaceutical Biotechnology, 66123 Saarbrücken, Germany.
authors.forename: Andriy
authors.initials: A
authors.lastname: Luzhetskyy
authors.affiliation: Institute of Molecular Biology, Slovak Academy of Sciences , 845 51 Bratislava, Slovak Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Kormanec
authors.affiliation: Department of Pharmacognosy, University of Vienna , 1090 Vienna, Austria.
authors.forename: Olga N
authors.initials: ON
authors.lastname: Sekurova
authors.affiliation: Department of Pharmacognosy, University of Vienna , 1090 Vienna, Austria.
authors.forename: Sergey B
authors.identifier: 0000-0002-9324-245X
authors.initials: SB
authors.lastname: Zotchev
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D055808:Drug Discovery / Q000379:methods; D055442:Metabolome; D005810:Multigene Family / Q000235:genetics*; D011353:Prodigiosin / Q000031:analogs & derivatives; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C528938:undecylprodiginine; D011353:Prodigiosin
keywords: Streptomyces; TetR repressor; biosensor; orphan gene clusters; secondary metabolite biosynthesis