Abstract：
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.
摘要：
聚对苯二甲酸乙二醇酯(PET)已引起严重的污染问题。与高能耗和成本的化学降解相比，酶降解为PET废物回收提供了可持续的解决方案。然而,目前PET水解酶的水解活性仍需要改进。在这项研究中,开发了一种基于互相关的累积诱变（CAM）策略来增强水解活性。通过减轻上位效应和组合突变，我们在LCC-ICCG的无定形PET膜上获得了具有2.1倍水解活性的高活性变体LCC-YGA(H183Y/L124G/S29A)。构象分析阐明了引入远端突变如何增强活性。不同地区之间的动态相关性促进了协同效应，通过远程交互增强绑定口袋的灵活性。完全正确,这项工作为PET水解酶工程提供了新的见解和方法，并为PET降解和回收提供了一种有效的酶。
