文献类型：期刊文献

英文题名：Break Through the Thermostability of Glucose Oxidase in Extremely Thermal Environments with a Novel Dynamic Ensemble Design Protocol

作者：Zhang, Lujia[1,3]; Miao, Tingwei[1]; Zhi, Fengdong[1]; Yang, Xin[1]; Yuan, Zhaoting[1]; Zhang, Chuanxi[4]; Feng, Yinghui[1]; Wei, Hao[4]; Gao, Bei[2]

机构：[1] Shanghai Engineering Research Center of Molecular Therapeutics & New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai, 200062, China; [2] School of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China; [3] NYU-ECNU Center for Computational Chemistry at NYU Shanghai, Shanghai, 200062, China; [4] Department of Micro/Nano Electronics, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China

年份：2024

外文期刊名：SSRN

收录：EI(收录号：20240240751)

语种：英文

外文关键词：Computational methods - Design - Economic and social effects - Enzyme activity - Glucose - Glucose oxidase - Glucose sensors - Molecular dynamics

摘要：Enhancing the thermostability of glucose oxidase (GOD) is crucial for food science applications. However, traditional design methods based on a single GOD structure are ineffective in extreme thermal environments. Here, we established a method for precisely locating residue by analysing the dynamic conformations of GoxM8 (M8) and further enhancing thermostability while maintaining activity. Our novel dynamic ensemble approach, coupled with FireProt computational analyses, was used to obtain the best mutant, V402F, from diverse conformations of M8. V402F residual activity was six times that of M8 at 80 °C for 2 min, and enzyme activity was even slightly increased. Experimental validation and computational analysis of stability mechanisms demonstrated the deficiencies of previous design strategies for flexible enzymes, proving the validity of our approach. Thus, we present a GOD variant with improved thermostability, as well as a more precise and efficient design strategy for GOD and other flexible enzymes. ? 2024, The Authors. All rights reserved.