Enhanced Poly(ethylene terephthalate) Hydrolase Activity by Protein Engineering

Yuan Ma; Mingdong Yao; Bingzhi Li; Mingzhu Ding; Bo He; Si Chen; Xiao Zhou; Yingjin Yuan

doi:10.1016/j.eng.2018.09.007

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Engineering ›› 2018, Vol. 4 ›› Issue (6) : 888-893. DOI: 10.1016/j.eng.2018.09.007

Research

Research Synthetic Biology—Article

Enhanced Poly(ethylene terephthalate) Hydrolase Activity by Protein Engineering

Yuan Ma^a^,^b ,
Mingdong Yao^a^,^b ,
Bingzhi Li^a^,^b ,
Mingzhu Ding^a^,^b ,
Bo He^a^,^b ,
Si Chen^a^,^b ,
Xiao Zhou^a^,^b ,
Yingjin Yuan^a^,^b

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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.

Keywords

Polyesterase / PET degradation / Cell-free protein synthesis / Polyester / PETase

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Yuan Ma, Mingdong Yao, Bingzhi Li, Mingzhu Ding, Bo He, Si Chen, Xiao Zhou, Yingjin Yuan. Enhanced Poly(ethylene terephthalate) Hydrolase Activity by Protein Engineering. Engineering, 2018, 4(6): 888‒893 https://doi.org/10.1016/j.eng.2018.09.007

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Acknowledgements

This work was funded by 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), and the Innovative Talents and Platform Program of Tianjin (16PTGCCX00140 and 16PTSYJC00050).

Compliance with ethics guidelines

Yuan Ma, Mingdong Yao, Bingzhi Li, Mingzhu Ding, Bo He, Si Chen, Xiao Zhou, and Yingjin Yuan declare that they have no conflict of interest or financial conflicts to disclose.