文献类型：期刊文献

英文题名：Serine 85 functions as a catalytic acid in the reprotonation process during EvAS-catalyzed astellifadiene biosynthesis

作者：Zhang, Weiyan[1];Peng, Kaitong[1];Lan, Keying[1];Xu, Kangwei[2];Wu, Ruibo[2];Hsiang, Tom[3];Nie, Shaoping[4];Zhang, Lixin[1];Wang, Xinye[1,5];Liu, Xueting[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Sun Yat Sen Univ, Sch Pharmaceut Sci, Guangzhou 510006, Guangdong, Peoples R China;[3]Univ Guelph, Sch Environm Sci, 50 Stone Rd East, Guelph, ON N1G 2W1, Canada;[4]Nanchang Univ, State Key Lab Food Sci & Technol, Nanchang 330031, Jiangxi, Peoples R China;[5]Ludong Univ, Sch Life Sci, Yantai 264025, Shandong, Peoples R China

年份：2024

卷号：60

期号：80

起止页码：11319

外文期刊名：CHEMICAL COMMUNICATIONS

收录：;EI(收录号：20243917084928);WOS:【SCI-EXPANDED(收录号:WOS:001315008400001)】；

基金：This work was supported by the National Key Research and Development Program of China (2019YFA0906201, 2020YFA090032 and 2022YFC2105400), the National Natural Science Foundation of China China (21977029, 81903529, 31720103901, 22307037), the 111 Project (B18022), the Science and Technology Commission of Shanghai Municipality (21NL2600100), and the Open Project Funding of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Biosynthesis - Catalysis

摘要：The deprotonation-reprotonation sequence introduces additional cyclization branches in terpene biosynthesis. However, the underlying mechanism remains poorly understood. In this study, we employed a combined approach of molecular dynamics (MD) simulations and site-directed mutagenesis on astellifadiene synthase EvAS from Emericella variecolor to investigate the role of a protonated S85 residue. This residue acts as a catalytic acid, previously unreported, that facilitates the reprotonation step in astellifadiene biosynthesis. Mutating S85 led to the production of a new tricyclic sesterterpene. A joint computational chemistry and site-directed mutagenesis approach identified a protonated S85 residue as the catalytic acid in astellifadiene synthase EvAS.