文献类型：期刊文献

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

作者：Shi, Tong[1,2];Zhang, Lu[1,2];Liang, Mindong[1,2];Wang, Weishan[3,4];Wang, Kefeng[1,2];Jiang, Yue[1,2];Liu, Jing[1,2];He, Xinwei[1,2];Yang, Zhiheng[1,2];Chen, Haihong[1,2];Li, Chuan[1,2];Lv, Dongyuan[1,2];Zhou, Liming[1,2];Chen, Biqin[5];Li, Dan[5];Zhang, Li-Xin[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]Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Resources, Beijing 100101, Peoples R China;[4]Chinese Acad Sci, Inst Microbiol, CAS Key Lab Pathogen Microbiol & Immunol, Beijing 100101, Peoples R China;[5]Inner Mongolia Kingdomway Pharmaceut Co Ltd, Hohhot 010206, Peoples R China

年份：2021

卷号：6

期号：4

起止页码：335

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000751846300003)】；

基金：This work was supported by the National Natural Science Foundation of China [31870040] , the National Key Research and Development Project (2020YFA0907804, 2020YFA0907304) , the "111" Project of China [B18022] , the Fundamental Research Funds for the Central Uni-versities [22221818014] , and the Open Project Funding of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Rhodobacter sphaeroides; Promoter library; Transcriptomics; Co-enzyme Q 10; Red fluorescent protein

摘要：The versatile photosynthetic alpha-proteobacterium Rhodobacter sphaeroides, has recently been extensively engineered as a novel microbial cell factory (MCF) to produce pharmaceuticals, nutraceuticals, commodity chemicals and even hydrogen. However, there are no well-characterized high-activity promoters to modulate gene transcription during the engineering of R. sphaeroides. In this study, several native promoters from R. sphaeroides JDW-710 (JDW-710), an industrial strain producing high levels of co-enzyme Q10 (Q10) were selected on the basis of transcriptomic analysis. These candidate promoters were then characterized by using gusA as a reporter gene. Two 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 R. sphaeroides. In addition, a Prsp_7571derived 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 R. sphaeroides. Finally, 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/L. These well-characterized promoters should be highly useful in current synthetic biology platforms for refactoring the biosynthetic pathway in R. sphaeroides-derived MCFs.