Enhanced Poly(ethylene terephthalate)Hydrolase Activity by Protein Engineering  

作　　者：Yuan Ma Mingdong Yao Bingzhi Li Mingzhu Ding Bo He Si Chen Xiao Zhou Yingjin Yuan 

机构地区：[1]Key Laboratory of Systems Bioengineering(Ministry of Education),School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China [2]SynBio Research Platform,Collaborative Innovation Center of Chemical Science and Engineering,Tianjin University,Tianjin 300072,China

出　　处：《Engineering》2018年第6期888-893,共6页工程（英文）

基　　金：National Program on Key Basic Research Project by the Ministry of Science and Technology of China(2014CB745100);the National Natural Science Foundation of China(21676190 and 21621004);the Innovative Talents and Platform Program of Tianjin(16PTGCCX00140 and 16PTSYJC00050).

摘　　要：Poly(ethylene terephthalate)hydrolase(PETase)from Ideonella sakaiensis exhibits a strong ability to degrade poly(ethylene terephthalate)(PET)at room temperature,and is thus regarded as a potential tool to solve the issue of polyester plastic pollution.Therefore,we explored the interaction between PETase and the substrate(a dimer of the PET monomer ethylene terephthalate,2PET),using a model of PETase and its substrate.In this study,we focused on six key residues around the substrate-binding groove in order to create novel high-efficiency PETase mutants through protein engineering.These PETase mutants were designed and tested.The enzymatic activities of the R61A,L88F,and I179F mutants,which were obtained with a rapid cell-free screening system,exhibited 1.4 fold,2.1 fold,and 2.5 fold increases,respectively,in comparison with wild-type PETase.The I179F mutant showed the highest activity,with the degradation rate of a PET film reaching 22.5 mg perμmol·L^-1 PETase per day.Thus,this study has created enhanced artificial PETase enzymes through the rational protein engineering of key hydrophobic sites,and has further illustrated the potential of biodegradable plastics.

关 键 词：Polyesterase PET DEGRADATION CELL-FREE PROTEIN synthesis POLYESTER PETase 

分 类 号：N[自然科学总论]