文献类型：期刊文献

英文题名：Promoter screening and identification for metabolic regulation in Acremonium chrysogenum

作者：Liu, Ling[1,2];Li, Runge[1];Zhang, Xinrun[1];Chen, Zhen[1];Mohsin, Ali[1];Hang, Haifeng[1,2];Tian, Xiwei[1,2,3];Chu, Ju[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Qingdao Innovat Inst, Qingdao, Peoples R China;[3]POB 329,130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2024

卷号：19

期号：3

外文期刊名：BIOTECHNOLOGY JOURNAL

收录：;EI(收录号：20241215771124);WOS:【SCI-EXPANDED(收录号:WOS:001184534700001)】；

基金：This work was financially supported by the National Key Research Development Program of China (2021YFC2100203), the Taishan Scholars Program, the Natural Science Foundation of Shanghai (Grant No. 23ZR1416500), and the Frontiers Science Center for Materiobiology and Dynamic Chemistry (JKVJ1231036).

语种：英文

外文关键词：Acremonium chrysogenum; bidirectional promoter; cephalosporin C; Golden Gate assembly; promoter library

摘要：Acremonium chrysogenum is the major industrial producer of cephalosporin C (CPC), which is used as raw material for the production of significant cephalosporin antibiotics. Due to the lack of diverse promoter elements, the development of metabolic engineering transformation is relatively slow, resulting in a limited improvement on CPC production. In this study, based on the analysis of the transcriptome profile, 27 candidate promoters were selected to drive the expression of the reporter genes. The promoter activities of this library ranged from 0.0075 to 101 times of the control promoter P-AngpdA. Simultaneously, a rapid screening method for potential bidirectional promoters was developed and 4 strong bidirectional promoters from 27 candidate options were identified and validated. Finally, the Golden Gate method was employed to combine promoter modules from the library with various target genes. Through a mixed transformation and screening process, high-yielding strains AG-6, AG-18, and AG-41 were identified, exhibiting an increase in CPC production of 30%, 35%, and 29%, respectively, compared to the control strain Ac-triangle axl2:: eGFP. Therefore, the utilization of this promoter library offers a broader range of synthetic biology toolkits for the genetic engineering transformation of A. chrysogenum, thus establishing a solid foundation for the precise regulation of gene expression.