文献类型：期刊文献

英文题名：Characterizing Y224 conformational flexibility in FtmOx1-catalysis using 19F NMR spectroscopy

作者：Wang, Xinye[1];Yang, Lingyun[2];Wang, Shenlin[1];Wang, Jun[3];Li, Kelin[4];Naowarojna, Nathchar[5];Ju, Yi[1];Ye, Ke[1];Han, Yuchen[1];Yan, Wupeng[3];Liu, Xueting[1];Zhang, Lixin[1];Liu, Pinghua[4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China;[3]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China;[4]Boston Univ, Dept Chem, Boston, MA 02215 USA;[5]Sakon Nakhon Rajabhat Univ, Fac Sci & Technol, Program Chem, Sakon Nakhon 47000, Thailand

年份：2025

卷号：15

期号：2

起止页码：386

外文期刊名：CATALYSIS SCIENCE & TECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001372796600001)】；

基金：This work was supported by the National Key Research and Development Program of China (2020YFA0907800 and 2022YFC2105400), the National Natural Science Foundation of China (22307037, 31720103901, 21977029, 81903529, and 32101008), the Open Project Funding of the State Key Laboratory of Bioreactor Engineering, and the 111 Project (B18022). This work was supported in part by the National Institutes of Health (GM140040 to P. L.). The authors thank the staff members at the BL02U1 and BL19U1 beamline at the Shanghai Synchrotron Radiation Facility (SSRF), China, for their assistance in X-ray crystal data collection. A CD spectrometer was provided by NSF MRI grant CHE-112654.

语种：英文

摘要：alpha-Ketoglutarate-dependent non-haem iron (alpha KG-NHFe) enzymes play a crucial role in natural product biosynthesis, and in some cases exhibiting multifunctional catalysis capability. This study focuses on alpha KG-NHFe enzyme FtmOx1, which catalyzes endoperoxidation, dealkylation, and alcohol oxidation reactions in verruculogen biosynthesis. We explore the hypothesis that the conformational dynamics of the active site Y224 confer the multifunctional activities of FtmOx1-catalysis. Utilizing Y224-to-3,5-difluorotyrosine-substituted FtmOx1, produced via the amber codon suppression method, we conducted 19F NMR characterization to investigate FtmOx1's structural flexibility. Subsequent biochemical and X-ray crystallographic analyses provided insights into how specific conformations of FtmOx1-substrate complexes influence their catalytic activities. These findings underscore the utility of 19F NMR as a powerful tool for elucidating the complex mechanisms of multifunctional enzymes, offering potential avenues for developing biocatalytic processes to produce novel therapeutic agents harnessing their unique catalytic properties.