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

作　　者：Xinye Wang Yiyi Huang Weiyan Zhang Kangjie Lv Xiaoying Li Zhixin Wang Li Zhang Tom Hsiang Lixin Zhang Liming Ouyang Xueting Liu 

机构地区：[1]State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai,200237,China [2]Department of Chemistry,Boston University,Boston,MA,USA [3]School of Environmental Sciences,University of Guelph,50 Stone Road East,Guelph,Ontario,N1G 2W1,Canada

出　　处：《Synthetic and Systems Biotechnology》2024年第2期380-387,共8页合成和系统生物技术（英文）

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).

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.

关 键 词：Bifunctional terpene synthase Sesterterpene Carbocation transportation Density functional theory calculations Molecular dynamics simulations 

分 类 号：O64[理学—物理化学]