title: Characterization of the polyoxin biosynthetic gene cluster from Streptomyces cacaoi and engineered production of polyoxin H.
abstract: A gene cluster (pol) essential for the biosynthesis of polyoxin, a nucleoside antibiotic widely used for the control of phytopathogenic fungi, was cloned from Streptomyces cacaoi. A 46,066-bp region was sequenced, and 20 of 39 of the putative open reading frames were defined as necessary for polyoxin biosynthesis as evidenced by its production in a heterologous host, Streptomyces lividans TK24. The role of PolO and PolA in polyoxin synthesis was demonstrated by in vivo experiments, and their functions were unambiguously characterized as O-carbamoyltransferase and UMP-enolpyruvyltransferase, respectively, by in vitro experiments, which enabled the production of a modified compound differing slightly from that proposed earlier. These studies should provide a solid foundation for the elucidation of the molecular mechanisms for polyoxin biosynthesis, and set the stage for combinatorial biosynthesis using genes encoding different pathways for nucleoside antibiotics.
authors.affiliation: Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200030, China.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Chen
authors.forename: Tingting
authors.initials: T
authors.lastname: Huang
authors.forename: Xinyi
authors.initials: X
authors.lastname: He
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Meng
authors.forename: Delin
authors.initials: D
authors.lastname: You
authors.forename: Linquan
authors.initials: L
authors.lastname: Bai
authors.forename: Jialiang
authors.initials: J
authors.lastname: Li
authors.forename: Mingxuan
authors.initials: M
authors.lastname: Wu
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.forename: Zhoujie
authors.initials: Z
authors.lastname: Xie
authors.forename: Huchen
authors.initials: H
authors.lastname: Zhou
authors.forename: Xiufen
authors.initials: X
authors.lastname: Zhou
authors.forename: Huarong
authors.initials: H
authors.lastname: Tan
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
mesh_terms: D002874:Chromosome Mapping; D005816:Genetic Complementation Test; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family; D016296:Mutagenesis; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D011741:Pyrimidine Nucleosides / Q000096:biosynthesis / Q000235:genetics; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
chemical_list: D020033:Protein Isoforms; D011741:Pyrimidine Nucleosides; C002457:polyoxin