文献类型：期刊文献

英文题名：Heterologous Biosynthesis of Spinosad: An Omics-Guided Large, Polyketide Synthase Gene Cluster Reconstitution in Streptomyces

作者：Tan, Gao-Yi[1,2,4];Deng, Kunhua[1,2,3];Liu, Xinhua[1,2,3];Tao, Hui[1,2,3];Chang, Yingying[1,2,3];Chen, Jia[1,2,3];Chen, Kai[5];Sheng, Zhi[5];Deng, Zixin[1,2,3,6,7];Liu, Tiangang[1,2,3]

机构：[1]Wuhan Univ, Key Lab Combinatorial Biosynth & Drug Discovery, Minist Educ, Wuhan 430071, Peoples R China;[2]Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Peoples R China;[3]Wuhan Inst Biotechnol, Hubei Engn Lab Synthet Microbiol, Wuhan 430075, Peoples R China;[4]East China Univ Sci & Technol, Sch Bioengn, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[5]Shenyang Res Inst Chem Ind Ltd Co, SINOCHEM Grp, Shenyang 110021, Peoples R China;[6]Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, 800 Dongchuan Rd, Shanghai 200240, Peoples R China;[7]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, 800 Dongchuan Rd, Shanghai 200240, Peoples R China

年份：2017

卷号：6

期号：6

起止页码：995

外文期刊名：ACS SYNTHETIC BIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000403864900009)】；

基金：We thank Prof. Meizhong Luo (National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University) for his help in the construction of the BAC library. We also thank Shuai Fu (J1 Biotech Co. Ltd, Wuhan, China) and Dr. Xiaoyan Xu (AB Sciex Pte. Ltd) for her help in proteomics and metabolomics analysis, respectively. This study was supported by National 973 Program of China (No. 2012CB721000), the Open Project of State Key Laboratory of Microbial Metabolism funded this work (No. MMLKF15-13), the Science and Technology Investment Program of SINOCHEM Group (No. 2015ADSW0039), and the Young Talents Program of National High-level Personnel of Special Support Program (The "Ten Thousand Talent Program") to T. Liu. The authors also specially acknowledge the support from the National Natural Science Foundation of China (No. 31500072), the Natural Science Foundation of HuBei Province (No. 2015CFB415), and China Postdoctoral Science Foundation Grant (No. 2014M562052).

语种：英文

外文关键词：spinosyn; Streptornyces; heterologous production; metabolomics; proteomics; module optimization

摘要：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 Streptornyces 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 mu g/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.