Engineering the thermostability of D-lyxose isomerase from Caldanaerobius polysaccharolyticus via multiple computer-aided rational design for efficient synthesis of D-mannose

作     者：Hao Wu Ming Yi Xiaoyi Wu Yating Ding Minghui Pu Li Wen Yunhui Cheng Wenli Zhang Wanmeng Mu 

作者机构：School of Food Science and BioengineeringChangsha University of Science&TechnologyChangsha410114China State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxi214122China 

出 版 物：《Synthetic and Systems Biotechnology》 (合成和系统生物技术（英文）)

年 卷 期：2023年第8卷第2期

页      面：323-330页

核心收录：

学科分类：0831[工学-生物医学工程（可授工学、理学、医学学位）] 0710[理学-生物学] 081704[工学-应用化学] 1001[医学-基础医学(可授医学、理学学位)] 07[理学] 08[工学] 0817[工学-化学工程与技术] 070303[理学-有机化学] 0703[理学-化学] 0836[工学-生物工程] 

基　　金：supported by the National Natural Science Foundation of China(32201963) Scientific Research Foundation of Hunan Provincial Education Department(22C0137) 

主　　题：D-lyxose isomerase Thermostability D-mannose Molecular modification 

摘      要：D-Mannose is an attractive functional sugar that exhibits many physiological benefits on human *** demand for low-calorie sugars and sweeteners in foods are increasingly available on the *** sugar isomerases,such as D-lyxose isomerase(D-LIase),can achieve an isomerization reaction between D-mannose and ***,the weak thermostability of D-LIase limits its efficient conversion from D-fructose to ***,few studies are available that have investigated the molecular modification of D-LIase to improve its thermal *** this study,computer-aided tools including FireProt,PROSS,and Consensus Finder were employed to jointly design D-LIase mutants with improved thermostability for the first ***,the obtained five-point mutant M5(N21G/E78P/V58Y/C119Y/K170P)showed high thermal stability and cat-alytic *** half-life of M5 at 65◦C was 10.22 fold,and the catalytic efficiency towards 600 g/L of D-fructose was 2.6 times to that of the wild type enzyme,*** dynamics simulation and intramolecular forces analysis revealed a thermostability mechanism of highly rigidity conformation,newly formed hydrogen bonds andπ-cation interaction between and within protein domains,and redistributed surface electrostatic charges for the mutant *** research provided a promising D-LIase mutant for the industrial production of D-mannose from D-fructose.