title: Heterologous Biosynthesis of Spinosad: An Omics-Guided Large Polyketide Synthase Gene Cluster Reconstitution in Streptomyces.
abstract: With the advent of the genomics era, heterologous gene expression has been used extensively as a means of accessing natural products (NPs) from environmental DNA samples. However, the heterologous production of NPs often has very low efficiency or is unable to produce targeted NPs. Moreover, due to the complicated transcriptional and metabolic regulation of NP biosynthesis in native producers, especially in the cases of genome mining, it is also difficult to rationally and systematically engineer synthetic pathways to improved NPs biosynthetic efficiency. In this study, various strategies ranging from heterologous production of a NP to subsequent application of omics-guided synthetic modules optimization for efficient biosynthesis of NPs with complex structure have been developed. Heterologous production of spinosyn in Streptomyces spp. has been demonstrated as an example of the application of these approaches. Combined with the targeted omics approach, several rate-limiting steps of spinosyn heterologous production in Streptomyces spp. have been revealed. Subsequent engineering work overcame three of selected rate-limiting steps, and the production of spinosad was increased step by step and finally reached 1460 μg/L, which is about 1000-fold higher than the original strain S. albus J1074 (C4I6-M). These results indicated that the omics platform developed in this work was a powerful tool for guiding the rational refactoring of heterologous biosynthetic pathway in Streptomyces host. Additionally, this work lays the foundation for further studies aimed at the more efficient production of spinosyn in a heterologous host. And the strategy developed in this study is expected to become readily adaptable to highly efficient heterologous production of other NPs with complex structure.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology , 130 Meilong Road, Shanghai 200237, China.
authors.forename: Gao-Yi
authors.initials: GY
authors.lastname: Tan
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Kunhua
authors.initials: K
authors.lastname: Deng
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Xinhua
authors.initials: X
authors.lastname: Liu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Tao
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.affiliation: Shenyang Research Institute of Chemical Industry Ltd., Co. , SINOCHEM Group, Shengyang 110021, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Chen
authors.affiliation: Shenyang Research Institute of Chemical Industry Ltd., Co. , SINOCHEM Group, Shengyang 110021, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Sheng
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.|State Key Laboratory of Microbial Metabolism, and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University , 800 Dongchuan Road, Shanghai 200240, China.
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology , Wuhan 430075, China.
authors.forename: Tiangang
authors.identifier: 0000-0001-8087-0345
authors.initials: T
authors.lastname: Liu
mesh_terms: D019149:Bioreactors / Q000382:microbiology; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D004338:Drug Combinations; D015723:Gene Library; D018942:Macrolides / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D055432:Metabolomics; D005810:Multigene Family / Q000235:genetics*; D048630:Polyketide Synthases / Q000235:genetics* / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D058615:Synthetic Biology
chemical_list: D004338:Drug Combinations; D018942:Macrolides; D048630:Polyketide Synthases; C415329:spinosad
keywords: Streptomyces; heterologous production; metabolomics; module optimization; proteomics; spinosyn