文献类型：期刊文献

英文题名：Expanding catalytic promiscuity of a bifunctional terpene synthase through a single mutation-induced change in hydrogen-bond network within the catalytic pocket

作者：Wang, Xinye[1];Huang, Yiyi[1];Zhang, Weiyan[1];Lv, Kangjie[1];Li, Xiaoying[1];Wang, Zhixin[1];Zhang, Li[2];Hsiang, Tom[3];Zhang, Lixin[1];Ouyang, Liming[1];Liu, Xueting[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Boston Univ, Dept Chem, Boston, MA USA;[3]Univ Guelph, Sch Environm Sci, 50 Stone Rd East, Guelph, ON N1G 2W1, Canada

年份：2024

卷号：9

期号：2

起止页码：380

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;EI(收录号：20240053712);WOS:【SCI-EXPANDED(收录号:WOS:001225661800001)】；

基金：We gratefully acknowledge the financial support from the National Key Research and Development Program of China (2019YFA0906201, 2020YFA090032, 2022YFC2105400) , the National Natural Science Foundation of China (22307037, 21907031, 81903529, 21977029, 31720103901, 21877124) , the Open Project Funding of the State Key Laboratory of Bioreactor Engineering, the 111 Project (B18022) .

语种：英文

外文关键词：Bifunctional terpene synthase; Sesterterpene; Carbocation transportation; Density functional theory calculations; Molecular dynamics simulations

摘要：Fungal bifunctional terpene synthases (BFTSs) catalyze the formation of numerous di-/sester-/tri-terpenes skeletons. However, the mechanism in controlling the cyclization pattern of terpene scaffolds is rarely deciphered for further application of tuning the catalytic promiscuity of terpene synthases for expanding the chemical space. In this study, we expanded the catalytic promiscuity of Fusarium oxysporum fusoxypene synthase (FoFS) by a single mutation at L89, leading to the production of three new sesterterpenes. Further computational analysis revealed that the reconstitution of the hydrogen-bond (H-bond) network of second-shell residues around the active site of FoFS influences the orientation of the aromatic residue W69 within the first-shell catalytic pocket. Thus, the dynamic orientation of W69 alters the carbocation transport, leading to the production of diverse ring system skeletons. These findings enhance our knowledge on understanding the molecular mechanisms, which could be applied on protein engineering terpene synthases on regulating the terpene skeletons.