〔Abstract〕 Objective To prepare a recombinant PETase with a dual-catalytic-triad and to evaluate its efficiency in the biodegradation of polyethylene terephthalate （PET）. Methods Based on the crystal structure of wild-type PETase， point mutations （T88H/L117D） were introduced via site-directed mutagenesis. The recombinant protein was prepared using prokaryotic expression and chromatography purification techniques. The enzymatic hydrolysis of the mutant PETase was assessed by relatively quantifying the products mono （2-hydroxyethyl） terephthalate （MHET） and terephthalic acid （TPA）. Results Both wild-type and mutant PETases accumulated as inclusion bodies， accounting for approximately 20% of the total bacterial protein. After solubilization in urea， the proteins were eluted at 300 mmol/L imidazole during affinity chromatography purification， with concentrations of 1. 824 and 1. 833 mg/mL and purities of 83. 11% and 84. 32%， respectively. Subsequent anion-exchange chromatography yielded highly pure enzymes in the 200 mmol/L NaCl fraction：2. 776 mg/mL （96. 86% purity） for the wild type and 1. 967 mg/mL （95. 13% purity） for the mutant. Following refolding， the final concentrations were 0. 484 mg/mL for the wild type and 0. 991 mg/mL for the mutant. Hydrolysis assays revealed that the mutant released MHET and TPA at （237. 67±17. 00）% and （197. 33±12. 01）% of the wild-type levels， respectively. Conclusion The T88H/L117D dual-catalytic-triad PETase is successfully prepared and it significantly enhanced PET-degrading ac‑ tivity， thus， it′s a promising biocatalyst for PET bioremediation.