Screening and engineering of high-activity promoter elements through transcriptomics and red fluorescent protein visualization in Rhodobacter sphaeroides

作     者：Tong Shi Lu Zhang Mindong Liang Weishan Wang Kefeng Wang Yue Jiang Jing Liu Xinwei He Zhiheng Yang Haihong Chen Chuan Li Dongyuan Lv Liming Zhou Biqin Chen Dan Li Li-Xin Zhang Gao-Yi Tan 

作者机构：State Key Laboratory of Bioreactor Engineering(SKLBE)And School of BiotechnologyEast China University of Science and Technology(ECUST)Shanghai200237China State Key Laboratory of Microbial Resources and CAS Key Laboratory of Pathogenic Microbiology and ImmunologyInstitute of MicrobiologyChinese Academy of Sciences(CAS)Beijing100101China Inner Mongolia Kingdomway Pharmaceutical Co.LtdHohhot010206China 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2021年第6卷第4期

页      面：335-342页

核心收录：

学科分类：081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 070302[理学-分析化学] 0703[理学-化学] 

基　　金：This work was supported by the National Natural Science Foundation of China the National Key Research and Development Project(2020YFA0907804,2020YFA0907304) the“111”Project of China[B18022] the Fundamental Research Funds for the Central Universities,and the Open Project Funding of the State Key Laboratory of Bioreactor Engineering 

主　　题：Rhodobacter sphaeroides Promoter library Transcriptomics Co-enzyme Q_(10) Red fluorescent protein 

摘      要：The versatile photosyntheticα-proteobacterium Rhodobacter sphaeroides,has recently been extensively engineered as a novel microbial cell factory(MCF)to produce pharmaceuticals,nutraceuticals,commodity chemicals and even ***,there are no well-characterized high-activity promoters to modulate gene transcription during the engineering of *** this study,several native promoters from *** JDW-710(JDW-710),an industrial strain producing high levels of co-enzyme Q10(Q10)were selected on the basis of transcriptomic *** candidate promoters were then characterized by using gusA as a reporter *** native promoters,Prsp_7571 and Prsp_6124,showed 620%and 800%higher activity,respectively,than the tac promoter,which has previously been used for gene overexpression in *** addition,a Prsp_7571-derived synthetic promoter library with strengths ranging from 54%to 3200%of that of the tac promoter,was created on the basis of visualization of red fluorescent protein(RFP)expression in ***,as a demonstration,the synthetic pathway of Q10 was modulated by the selected promoter T334*in JDW-710;the Q10 yield in shake-flasks increased 28%and the production reached 226 mg/*** well-characterized promoters should be highly useful in current synthetic biology platforms for refactoring the biosynthetic pathway in ***-derived MCFs.