文献类型：期刊文献

英文题名：Model-Guided Systematic Metabolic Engineering for Enhanced Spinosad Biosynthesis in Saccharopolyspora spinosa NHF132

作者：Wang, Shuliu[1,2];Liu, Yuxin[1,2];Zhang, Qian[1,2];Jiang, Yue[1,2];Zeng, Xiaoqian[1,2];Zhang, Chengyu[1,2];Cheng, Jiagao[1,2,3];Wang, Weishan[4];Zhang, Lixin[1,2];Tan, Gao-Yi[1,2]

机构：[1]East China Univ Sci & Technol ECUST, State Key Lab Bioreactor Engn SKLBE, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol ECUST, Sch Biotechnol, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab Chem Biol, Shanghai 200237, Peoples R China;[4]Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Divers & Innovat Utilizat, Beijing 100101, Peoples R China

年份：2025

卷号：12

期号：47

外文期刊名：ADVANCED SCIENCE

收录：;EI(收录号：20254119284972);WOS:【SCI-EXPANDED(收录号:WOS:001579028900001)】；

基金：This work was supported by the National Natural Science Foundation of China (32370064 and 32121005); the Shanghai Science and Technology Commission (24HC2820200); the 111 Project (B18022); Key Research & Development Program of Shandong Province (2022SFGC0104); Focus on Research and Development Plan in Shandong Province (2022CXGC020206), and the 2023 Double World-class Project-Key Program-Intelligent Biomanufacturing. The authors are grateful to Prof. Zaigao Tan at Shanghai Jiao Tong University for providing the pSET152-kasop*-BauA-SPL42-MCR-C plasmid containing the NCM pathway. The authors are thankful to Prof. Yinhua Lu at Shanghai Normal University for providing the pWT297 plasmid for genome editing in Sa. spinosa.

语种：英文

外文关键词：genome-scale metabolic model; Saccharopolyspora spinosa; secondary metabolite; spinosad; systems metabolic engineering

摘要：Spinosad (a mixture of spinosyns A and D) is a macrocyclic lactone green bioinsecticide produced by Saccharopolyspora spinosa. It is known for its high efficiency, low toxicity, and broad-spectrum activity. Although numerous strategies have been employed to enhance spinosad production, intricate regulation of secondary metabolism and inefficient genetic manipulation impede systematic and comprehensive metabolic engineering in this spinosad-producing strain. In this study, a genome-scale metabolic model (GEM) for Sa. spinosa NHF132 is developed to dissect the intricate secondary metabolic pathways of spinosad biosynthesis, analyzing interactions among precursors, key enzymes, and competing or bypass pathways. Guided by the model, the impact of rhamnose precursor overexpression, gene cluster amplification, short-chain acyl-CoA enhancement, and chassis optimization on spinosad production is systematically evaluated. By integrating these metabolic engineering strategies, engineered strain NHF132-BAC-SP43-NCM achieved a spinosad titer of 1816.8 mg L-1, a 553.3% increase over the starting strain, with substantial improvements in yield and product proportion. The model-driven framework for metabolic engineering of complex secondary metabolites in actinomycetes substantially increased spinosad production and offered valuable insights for other complex natural products.