文献类型：期刊文献

英文题名：Boosting the epoxidation of squalene to produce triterpenoids in Saccharomyces cerevisiae

作者：Du, Meng-Meng[1];Zhang, Ge-Ge[1];Zhu, Zhan-Tao[1];Zhao, Yun-Qiu[1];Gao, Bei[1];Tao, Xin-Yi[1];Wang, Feng-Qing[1];Wei, Dong-Zhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, POB 311, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2023

卷号：16

期号：1

外文期刊名：BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS

收录：;EI(收录号：20231914070428);WOS:【SCI-EXPANDED(收录号:WOS:000982152800001)】；

基金：This work was financially supported by the Natural Science Foundation of Shanghai (No.20ZR1415100), the National Natural Science Foundation of China (No. 21776075), and the National Natural Science Foundation of China (No. 21978084).

语种：英文

外文关键词：2,3-Oxidosqualene; 2,3:22,23-Dioxidosqualene; Triterpenoid; ERG7; Two-stage fermentation

摘要：Background Polycyclic triterpenoids (PTs) are common in plants, and have attracted considerable interest due to their remarkable biological activities. Currently, engineering the ergosterol synthesis pathway in Saccharomyces cerevisiae is a safe and cost-competitive way to produce triterpenoids. However, the strict regulation of ERG1 involved in the epoxidation of squalene limits the triterpenoid production. Results In this study, we found that the decrease in ERG7 protein level could dramatically boost the epoxidation of squalene by improving the protein stability of ERG1. We next explored the potential factors that affected the degradation process of ERG1 and confirmed that ERG7 was involved in the degradation process of ERG1. Subsequently, expression of four different triterpene cyclases utilizing either 2,3- oxidosqualene or 2,3:22,23-dioxidosqualene as the substrate in ERG7-degraded strains showed that the degradation of ERG7 to prompt the epoxidation of squalene could significantly increase triterpenoid production. To better display the potential of the strategy, we increased the supply of 2,3-oxidosqualene, optimized flux distribution between ergosterol synthesis pathway and ss-amyrin synthesis pathway, and modified the GAL-regulation system to separate the growth stage from the production stage. The best-performing strain ultimately produced 4216.6 +/- 68.4 mg/L of ss-amyrin in a two-stage fed-fermentation (a 47-fold improvement over the initial strain).