State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China; Collaborative Innovation Centre of Chemical Science and Engineering (Tianjin), Tianjin 300072, PR China; Tianjin Key Laboratory of Membrane Science and Desalination Technology, Tianjin University, Tianjin 300072, PR China.
Bioresour Technol. 2024 Aug;406:130929. doi: 10.1016/j.biortech.2024.130929. Epub 2024 Jun 3.
Polyethylene terephthalate (PET) has caused significant pollution issues. Compared to chemical degradation with high energy consumption and cost, enzymatic degradation offers a sustainable solution for PET waste recycling. However, the hydrolytic activity of current PET hydrolases still requires improvement. In this study, a cross-correlation-based accumulated mutagenesis (CAM) strategy was developed to enhance the hydrolysis activity. By mitigating epistatic effect and combinational mutations, we achieved a highly active variant LCC-YGA (H183Y/L124G/S29A) with 2.1-fold hydrolytic activity on amorphous PET films of LCC-ICCG. Conformational analysis elucidated how the introduction of distal mutations enhanced activity. The dynamic correlation among different regions facilitated a synergistic effect, enhancing binding pocket flexibility through remote interactions. Totally, this work offers novel insights and methods for PET hydrolases engineering and provides an efficient enzyme for PET degradation and recycling.
聚对苯二甲酸乙二醇酯（PET）造成了严重的污染问题。与高能耗和高成本的化学降解相比，酶降解为 PET 废物回收提供了一种可持续的解决方案。然而，目前 PET 水解酶的水解活性仍有待提高。在这项研究中，开发了一种基于互相关的累积诱变（CAM）策略来提高水解活性。通过减轻上位效应和组合突变，我们在无定形 PET 薄膜上实现了一个具有高活性的变体 LCC-YGA（H183Y/L124G/S29A），对 LCC-ICCG 的水解活性提高了 2.1 倍。构象分析阐明了引入远端突变如何增强活性。不同区域之间的动态相关性促进了协同作用，通过远程相互作用增强了结合口袋的灵活性。总的来说，这项工作为 PET 水解酶工程提供了新的见解和方法，并为 PET 的降解和回收提供了一种有效的酶。
