title: High Level of Spinosad Production in the Heterologous Host Saccharopolyspora erythraea.
abstract: Spinosad, a highly effective insecticide, has an excellent environmental and mammalian toxicological profile. Global market demand for spinosad is huge and growing. However, after much effort, there has been almost no improvement in the spinosad yield from the original producer, Saccharopolyspora spinosa Here, we report the heterologous expression of spinosad using Saccharopolyspora erythraea as a host. The native erythromycin polyketide synthase (PKS) genes in S. erythraea were replaced by the assembled spinosad gene cluster through iterative recombination. The production of spinosad could be detected in the recombinant strains containing the whole biosynthesis gene cluster. Both metabolic engineering and UV mutagenesis were applied to further improve the yield of spinosad. The final strain, AT-ES04PS-3007, which could produce spinosad with a titer of 830 mg/liter, has significant potential in industrial applications.
  
  
  
  This work provides an innovative and promising way to improve the industrial production of spinosad. At the same time, it also describes a successful method of heterologous expression for target metabolites of interest by replacing large gene clusters.
authors.affiliation: Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, China Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Huang
authors.affiliation: Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China.
authors.forename: Mei-Hong
authors.initials: MH
authors.lastname: Li
authors.affiliation: Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China.
authors.forename: Ji-Dong
authors.initials: JD
authors.lastname: Wang
authors.affiliation: Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Bai
authors.affiliation: Central Research Institute, Zhejiang Hisun Pharmaceutical Co., Ltd., Taizhou, China zhoujun@hisunpharm.com zhengyg@zjut.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, China zhoujun@hisunpharm.com zhengyg@zjut.edu.cn.
authors.forename: Yu-Guo
authors.initials: YG
authors.lastname: Zheng
mesh_terms: D003001:Cloning, Molecular; D004338:Drug Combinations; D015870:Gene Expression; D007306:Insecticides / Q000378:metabolism*; D018942:Macrolides / Q000378:metabolism*; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics; D005810:Multigene Family; D016930:Saccharopolyspora / Q000378:metabolism*
chemical_list: D004338:Drug Combinations; D007306:Insecticides; D018942:Macrolides; C415329:spinosad