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] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China; [2] iHuman Institute, Shanghaitech University, Shanghai, 201210, China; [3] School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China; [4] Department of Chemistry, Boston University, Boston, MA, 02215, United States; [5] Program of Chemistry, Faculty of Science and Technology, Sakon Nakhon Rajabhat University, Sakon Nakhon, 47000, Thailand

年份：2024

卷号：15

期号：2

起止页码：386

外文期刊名：Catalysis Science and Technology

收录：EI(收录号：20245117538244)

语种：英文

外文关键词：Biosynthesis - Complexation - Conformations - Nuclear magnetic resonance spectroscopy

摘要：α-Ketoglutarate-dependent non-haem iron (αKG-NHFe) enzymes play a crucial role in natural product biosynthesis, and in some cases exhibiting multifunctional catalysis capability. This study focuses on αKG-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-difluorotyrosinesubstituted 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. ? The Royal Society of Chemistry 2025.