title: Targeted activation of silent natural product biosynthesis pathways by reporter-guided mutant selection.
abstract: The continuously increasing genome sequencing data has revealed numerous cryptic pathways, which might encode novel secondary metabolites with interesting biological activities. However, utilization of this hidden potential has been hindered by the observation that many of these gene clusters remain silent (or poorly expressed) under laboratory conditions. Here we present reporter-guided mutant selection (RGMS) as an effective and widely applicable method for targeted activation of silent gene clusters in the native producers. The strategy takes advantage of genome-scale random mutagenesis for generation of genetic diversity and a reporter-guided selection system for the identification of the desired target-activated mutants. It was first validated in the re-activation of jadomycin biosynthesis in Streptomyces venezuelae ISP5230, where high efficiency of activation was achieved. The same strategy was then applied to a hitherto unactivable pga gene cluster in Streptomyces sp. PGA64 leading to the identification of two new anthraquinone aminoglycosides, gaudimycin D and E.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China; University of Chinese Academy of Sciences, Beijing 100049, People׳s Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China.
authors.forename: Sihai
authors.initials: S
authors.lastname: Xiang
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China.
authors.forename: Liyuan
authors.initials: L
authors.lastname: Li
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China; University of Chinese Academy of Sciences, Beijing 100049, People׳s Republic of China.
authors.forename: Bin
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, University of Turku, Vatselankatu 2, FIN-20014 Turku, Finland.
authors.forename: Johanna
authors.initials: J
authors.lastname: Rajasärkkä
authors.affiliation: Department of Biochemistry, University of Turku, Vatselankatu 2, FIN-20014 Turku, Finland.
authors.forename: Kirsi
authors.initials: K
authors.lastname: Gröndahl-Yli-Hannuksela
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China.
authors.forename: Guomin
authors.initials: G
authors.lastname: Ai
authors.affiliation: Department of Biochemistry, University of Turku, Vatselankatu 2, FIN-20014 Turku, Finland.
authors.forename: Mikko
authors.initials: M
authors.lastname: Metsä-Ketelä
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing 100101, People׳s Republic of China. Electronic address: yangkq@im.ac.cn.
authors.forename: Keqian
authors.initials: K
authors.lastname: Yang
mesh_terms: D000880:Anthraquinones / Q000378:metabolism; D005798:Genes, Bacterial; D017930:Genes, Reporter; D007546:Isoquinolines / Q000378:metabolism; D005810:Multigene Family; D009154:Mutation; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: D000880:Anthraquinones; D007546:Isoquinolines
keywords: Activation; Gaudimycin; Genome mining; RGMS; Silent gene cluster