Increasingγ-CD conversion rates by improving thermostability of Bacillus ***-44876γ-CGTase

作     者：Xiaoxiao Li Danni Zheng Jing Wu Zhengyu Jin Birte Svensson Yuxiang Bai 

作者机构：State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxiJiangsu214122China School of Food Science and TechnologyJiangnan UniversityWuxiJiangsu214122China Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu ProvinceJiangnan UniversityWuxiJiangsu214122China Enzyme and Protein ChemistryDepartment of Biotechnology and BiomedicineTechnical University of DenmarkDK-2800 KgsLyngbyDenmark 

出 版 物：《Food Bioscience》 (食品生物科学（英文）)

年 卷 期：2023年第51卷第1期

页      面：346-354页

核心收录：

学科分类：0817[工学-化学工程与技术] 08[工学] 

基　　金：supported by National Natural Science Foundation of China(No.32072268 and No.32201967) Natural Science Foundation of Jiangsu Province(BK20211581 and BK20221072) the International Joint Research Laboratory for Starch Related Enzymes at Jiangnan University with DTU Bioengineering 

主　　题：Starch Protein engineering Three-dimensional structure 

摘      要：Poor thermostability is a limiting factor for applications ofγ-CGTase that affects starch utilization and yield ofγ-*** thermostability of Bacillus ***-44876γ-CGTase(BFγ-CGTase)was improved by addition of Ca^(2+)and site-directed ***,10 mM Ca^(2+)increased the half-life(t1/2)at 55 and 60℃ from 3.0 to 0.3 h to 17.4 and 2.0 h,*** spectra indicated that Ca^(2+)stabilized the tertiary structure of the BFγ-CGTase and hence improved the *** to serine of a glycine residue in anα-helix related to thermostability also improved the stability especially at the higher ***,in 10 mM Ca^(2+)the G208S mutant further increased t1/2 to 20.0 h and 4.0 h at 55 and 60℃,*** G208S mutant in 10 mM Ca^(2+)producedγ-CD from tapioca starch with 40%increased yield of that from BFγ-*** work involving rational protein engineering provided a new tool for enzymaticγ-CD production.