标题:Enhanced Heterologous Spinosad Production from a 79-kb Synthetic Multioperon Assembly
作者:Song, Chaoyi; Luan, Ji; Cui, Qingwen; Duan, Qiuyue; Li, Zhen; Gao, Yunsheng; Li, Ruijuan; Li, Aiying; Shen, Yuemao; Li, Yuezhong; 更多
作者机构:[Song, Chaoyi; Luan, Ji; Cui, Qingwen; Duan, Qiuyue; Li, Zhen; Gao, Yunsheng; Li, Ruijuan; Li, Aiying; Shen, Yuemao; Li, Yuezhong; Zhang, Youming; Fu, 更多
通讯作者:Zhang, YM;Fu, J;Wang, HL
通讯作者地址:[Zhang, YM; Fu, J; Wang, HL]Shandong Univ, Sch Life Sci, Helmholtz Inst Biotechnol, State Key Lab Microbial Technol, Binhai Rd 72, Qingdao 266237, Peo 更多
来源:ACS SYNTHETIC BIOLOGY
出版年:2019
卷:8
期:1
页码:137-147
DOI:10.1021/acssynbio.8b00402
关键词:biosynthesis; gene clusters; synthetic operons; DNA engineering;; secondary metabolites; heterologous expression
摘要:Refactoring biosynthetic pathways for enhanced secondary metabolite production is a central challenge for synthetic biology. Here we applied advanced DNA assembly methods and a uniform overexpression logic using constitutive promoters to achieve efficient heterologous production of the complex insecticidal macrolide spinosad. We constructed a 79-kb artificial gene cluster in which 23 biosynthetic genes were grouped into 7 operons, each with a strong constitutive promoter. Compared with the original gene cluster, the artificial gene cluster resulted in a 328-fold enhanced spinosad production in Streptomyces albus J1074. To achieve this goal, we applied the ExoCET DNA assembly method to build a plasmid from 13 GC-rich fragments with high efficiency in one step. Together with our previous direct cloning and recombineering tools, we present new synthetic biology options for refactoring large gene clusters for diverse applications.
收录类别:SCOPUS;SCIE
资源类型:期刊论文
原文链接:https://www.scopus.com/inward/record.uri?eid=2-s2.0-85060163648&doi=10.1021%2facssynbio.8b00402&partnerID=40&md5=883b378ac23a7a00e0128abe2a1bef8c
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