Heterologous Biosynthesis and “Omics”-Guided Metabolic Engineering for Overproduction of Spinosad in Streptomyces

Authors: 
Tan, G. Y., Shenyang Research Institute of Chemical Industry Ltd., Co., SINOCHEM Group
Liu, X., Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Wuhan University School of Pharmaceutical Sciences
Deng, K., Wuhan University
Tao, H., Wuhan University
Liu, T., E-mail: liutg@whu.edu.cn
Deng, Z., Wuhan Institute of Biotechnology
Chen, K., Shenyang Research Institute of Chemical Industry Ltd., Co.
Li, X., Shenyang Research Institute of Chemical Industry Ltd., Co.

Spinosad, produced by Saccharopolyspora spinosa, is an important macrolide antibiotics with potent insecticidal properties. As a safe and environment-friendly pesticide, spinosad has been widely used in agriculture. However, due to the low productivity, the industrial production of spinosad is quite limited in China. In this study, a large size of 110kb engineered BAC plasmid, which harbored biosynthetic genes of Spinosad, has been introduced into Streptomyces host, such as Streptomyces albus J1074 and Streptomyces lividansTK24. Then, by RT-PCR analysis, the transcription of all the biosynthetic genes can be detected. In the flask fermentation experiment, the heterologous production of spinosad in Streptomyces host has been detected by Q-Exactive high-resolution mass spectrometer. To furtherly improved heterologous production of targeted product, the “omics”-guided metabolic engineering of spinosad production has been applied. By using proteomics and metabolomics approaches, several of rate-limiting steps of spinosad heterologous production have been revealed. The successful heterologous production in this work lays the foundation for overproduction of antibiotics, and future metabolic engineering work will be applied for high efficient production of spinosad in Streptomyces.