文献类型：期刊文献

英文题名：Rational spatial rewiring of key enzymes enhances α-santalene production in Saccharomyces cerevisiae

作者：Tan, Wenwen[1,2];Tong, Shengkun[2];Gao, Yaojie[2];Wang, Jing[1];Zhang, Jingyu[1,3];Xie, Zhiping[2];Dai, Huanqin[4,5];Liang, Yu[1,2];Tan, Gao-Yi[1];Zhang, Lixin[1,3];Tong, Yaojun[2]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab Joint Int Res Lab Me, Shanghai 200240, Peoples R China;[3]Shanghai Collaborat Innovat Ctr Biomfg Technol SCI, Shanghai 200237, Peoples R China;[4]Chinese Acad Sci, Inst Microbiol, State Key Lab Mycol, Beijing, Peoples R China;[5]Univ Chinese Acad Sci, Med Sch, Beijing 100049, Peoples R China

年份：2025

卷号：436

外文期刊名：BIORESOURCE TECHNOLOGY

收录：;EI(收录号：20253018853095);WOS:【SCI-EXPANDED(收录号:WOS:001542630000002)】；

基金：This work was supported by grants from the National Key Research and Development Program of China (2021YFA0909500, 2020YFA0907200, and 2020YFA0907800), the National Natural Science Foundation of China (32170080, 32370026, and 32121005), Shanghai Pilot Program for Basic Research-Shanghai Jiao Tong University (21TQ1400204), Science and Technology Commission of Shanghai Municipality (24HC2810200). Open Funding Project of the State Key Laboratory of Microbial Metabolism (MMLKF22-03), and the 111 Project (B18022).

语种：英文

外文关键词：Subcellular enzyme localization; Spatial engineering; alpha-Santalene; Saccharomyces cerevisiae; Metabolic engineering; Synthetic biology

摘要：Spatial compartmentalization in eukaryotic cell factories often constrains the efficiency of metabolic pathways. Here, we systematically mapped the subcellular localization of nine core enzymes in the alpha-santalene biosynthetic pathway of Saccharomyces cerevisiae, identifying metabolic bottlenecks associated with nuclear and endoplasmic reticulum (ER) localization. Through rational spatial engineering, including bioinformatically guided HMG1 truncation to achieve ER release and nuclear export signal (NES) tagging of key enzymes, we successfully rewired enzyme localization to enhance pathway flux. Coupled with promoter engineering to downregulate ERG9, addition-copy integration for IDI1 and ERG20 overexpression, and targeted medium optimization to improve cellular osmotolerance, we achieved substantial synergistic effects on production, leading to a 132-fold increase in alpha-santalene titer, reaching 568.59 mg/L in fed-batch fermentation. Our results demonstrate that combining subcellular localization engineering with classic metabolic and process optimization offers a robust and generalizable strategy for high-level terpenoid biosynthesis in S. cerevisiae. This approach not only advances the performance of S. cerevisiae cell factories but also holds promise for broader application across other yeast species and eukaryotic microbial hosts.