Enhanced Poly(ethylene terephthalate) Hydrolase Activity by Protein Engineering. (December 2018)

• Record Type: Journal Article
• Authors: Ma, Yuan; Yao, Mingdong; Li, Bingzhi; Ding, Mingzhu; He, Bo; Chen, Si; Zhou, Xiao; Yuan, Yingjin
• Is Part Of: Engineering. Volume 4:Number 6(2018)
• Page Start: 888
• Page End: 893
• Publication Date: 2018-12
• Subjects: Polyesterase -- PET degradation -- Cell-free protein synthesis -- Polyester -- PETase; Engineering -- Periodicals; Engineering -- China -- Periodicals; 620.005
• DOI: 10.1016/j.eng.2018.09.007
• Languages: English
• ISSNs: 2095-8099

Abstract

Abstract: 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 per μmol·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.