文献类型：期刊文献

英文题名：Harnessing respiratory chain engineering enhances erythromycin production in Saccharopolyspora erythraea employing chemically defined medium

作者：Jiang, Xing[1,2,3];Liu, Yazhi[1,2,3];Wang, Shuohan[1,2,3];Ke, Xiang[1,2,3];Shang, Qinghai[4];Li, Feng[4];Yu, Zhenhua[4];Li, Xu[1];Tian, Xiwei[1,2,3];Chu, Ju[1,2,3]

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;[4]HEC Pharm Co Ltd, Yichang 443300, Peoples R China

年份：2025

卷号：526

外文期刊名：CHEMICAL ENGINEERING JOURNAL

收录：;EI(收录号：20254719576347);WOS:【SCI-EXPANDED(收录号:WOS:001628515700011)】；

基金：This work was financially supported by National Key Research and Development Program of China (2022YFC2105403) , the Taishan Scholars Programs 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 Frontiers Science Center for Materiobiology and Dynamic Chemistry (JKVJ1231036) . We thank Professor Yinhua Lu (Shanghai Normal University, Shanghai, China) for providing pSET-dCas9-actII4-NT-S1.

语种：英文

外文关键词：Respiratory chain; Erythromycin; Saccharopolyspora erythraea; Terminal oxidase

摘要：The biosynthesis of erythromycin in Saccharopolyspora erythraea is highly energy-intensive, and the respiratory chain playing a key role in ATP generation and NADH oxidation. To understand the role of the respiratory chain in metabolic regulation and improve erythromycin production, the exploration of optimizing cellular energy metabolism and redox balance through respiratory chain engineering was carried out. In this study, a systematic investigation of respiratory chain components of S.erythraea was performed based on updated genomic annotations and literature data for the first time. Targeted gene suppression and overexpression were applied to identify the key elements essential for energy metabolism and erythromycin biosynthesis. Suppression of terminal oxidase and NDH-2 impaired growth and secondary metabolism, underscoring their role in NADH oxidation. Conversely, overexpression of respiratory chain genes allowed growth comparable to the parental strain. Among engineered strains, cytochrome bd oxidase overexpression (E3::cydABCD) was found to be the most effective. This strain increased erythromycin titer from 804 to 1017 mg/L in shake flasks culture, reduced the NADH/NAD+ ratio from 0.36 to 0.24, and elevated ATP levels from 98.6 to 145.4 mu M/gDCW. In a 5 L bioreactor, a titer of 2394 mg/L was achieved by E3::cydABCD after fermentation optimization-representing the highest reported titer using chemically defined medium. Metabolic flux analysis revealed enhanced energy and cofactor metabolism contributed to improved erythromycin production. This work demonstrates that respiratory chain engineering is an effective strategy to optimize redox balance and enhance industrial antibiotic production.