文献类型：期刊文献

英文题名：Enhancing Cephalosporin C Biosynthesis through a 2A Peptide-Based Multigene Coexpression System

作者：Chen, Zhen[1];Li, Yifan[1];Wei, Xiaorong[1];Hong, Wei[1];Guo, Yuanxin[1];Hang, Haifeng[1,2,3];Chu, Ju[1,2,3];Tian, Xiwei[1]

机构：[1]East China Univ Sci & Technol, Qingdao Innovat Inst, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Natl Ctr Bioengn & Technol Shanghai, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai 200237, Peoples R China

年份：2025

卷号：14

期号：11

起止页码：4547

外文期刊名：ACS SYNTHETIC BIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001600395100001)】；

基金：This work was financially supported by the National Key Research and Development Program of China (2024YFA0917703), the Taishan Scholars Program of Shandong Province (NO. tsqn202312316), the Shanghai Pilot Program for Basic Research (22TQ1400100-14), the Shanghai Science and Technology Innovation Action Plan (24HC2810100), the Natural Science Foundation of Shanghai (23ZR1416500), the Fundamental Research Funds for the Central Universities (JKV01251708). Thanks for the financial support from the Arawana Charity Foundation.

语种：英文

外文关键词：Acremonium chrysogenum; CPC; 2A peptide; multigene coexpression

摘要：Cephalosporin C (CPC) is a natural product that serves as the key precursor for various semisynthetic cephalosporins. Its industrial production primarily relies on Acremonium chrysogenum. However, as the exclusive microbial strain used for large-scale CPC fermentation, A. chrysogenum faces challenges in metabolic engineering owing to the absence of an efficient multigene coexpression system. This study presented, for the first time, the development and application of a 2A peptide-based multigene coexpression system in A. chrysogenum. The self-cleavage efficiencies of ten virus-derived 2A peptides were systematically evaluated, ranging from 66.5% to 88.2%, with P2A showing the best performance. The most efficient P2A peptide was then selected, enabling both high- and low-level precalibrated expression based on the transcriptome data and supporting the coexpression of three genes in A. chrysogenum, including the driver gene. This system was subsequently applied to coexpress the key CPC biosynthetic genes, cefEF and cefG, with the driver gene of ACRE_076110, resulting in a significant 3.19-fold increase in CPC titer compared to the wild-type strain. Furthermore, the strategy was successfully extended to an industrial high-yield strain, with CPC titer increasing from 6.09 g/L to 7.45 g/L, receiving a 22.2% improvement. Overall, this study provides a valuable tool for metabolic engineering efforts aimed at enhancing CPC production in A. chrysogenum.