a-Synuclein: A fusion chaperone significantly boosting the enzymaticperformance of PET hydrolase

作     者：Renwen Tian Yan Sun Renwen Tian;Yan Sun

作者机构：Department of Biochemical EngineeringSchool of Chemical Engineering and Technology and Key Laboratory of Systems Bioengineering and Frontiers Science Center for Synthetic Biology(Ministry of Education)Tianjin UniversityTianjin 300350China 

出 版 物：《Chinese Journal of Chemical Engineering》 (中国化学工程学报（英文版）)

年 卷 期：2023年第64卷第12期

页      面：18-25页

核心收录：

学科分类：081703[工学-生物化工] 08[工学] 0817[工学-化学工程与技术] 0804[工学-仪器科学与技术] 0805[工学-材料科学与工程（可授工学、理学学位）] 0703[理学-化学] 

基　　金：the National Key Research and Development Program of China(2018YFA0900702). 

主　　题：PET hydrolase a-Synuclein Fusion enzyme Stability Pollution Degradation 

摘      要：Extensive use of polyethylene terephthalate (PET) has brought about global environmental problems. Arecently reported PET hydrolase (PETase) discovered from Ideonella sakaiensis showed high potentialfor degrading PET at moderate temperatures, but its activity and stability need further improvementfor practical applications. Herein, we proposed to use a-synuclein (aS) as a fusion chaperone and createdsix PETase-aS fusion enzymes with linkers of different types and lengths. All the fusion enzymes exhibited improved enzymatic performance, presenting 1.5 to 2.6-fold higher activity towards bis-2(hydroxyethyl) terephthalate than PETase, as well as significantly increased stabilities. Fluorescencespectroscopy indicated that the chaperone fusion tightened the overall conformation and resulted inthe opening of the substrate binding pocket, which led to the improved thermal stability and catalyticactivity of the fusion enzymes. Remarkably, one of the fusion proteins, PETase-[(GS)(EK)]10-aS, showed3.2 to 5.1 times higher PET degradation capability than PETase. The significantly boosted PET degradationperformance was not only attributed to the enhanced enzymatic activity and stability, but also possiblydue to the binding affinity of the fused aS domain for PET. These findings demonstrated that aS was aneffective fusion chaperone for significantly enhancing the enzymatic performance of PETase.