Enhanced catalytic activity of polyethylene terephthalate hydrolase by structure-guided loop-focused iterative mutagenesis strategy

IF 12.2 1区环境科学与生态学 Q1 ENGINEERING, ENVIRONMENTAL Journal of Hazardous Materials Pub Date : 2025-03-05 DOI:10.1016/j.jhazmat.2025.137837

Wei Lin, Yunxin Zheng, Jiaxing Zhang, Yu Zhou, Mengfan Wang, Shengping You, Rongxin Su, Wei Qi

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Abstract

The accumulation of polyethylene terephthalate (PET) waste has caused significant environmental pollution. Although biological depolymerization offers a promising solution, its efficiency remains constrained by the limited activity of PET-degrading enzymes. In this study, we designed a Structure-guided Loop-focused Iterative Mutagenesis (SLIM) strategy and rationally engineered the PET degradation enzyme ICCG for higher activity. The strategy was designed by demonstrating the critical role of the β8-α6 loop in type Ⅰ enzymes, which has currently not been reported. The best variant obtained, YITA (H183Y/L202I/I208T/T153A), exhibited 4.46-fold higher hydrolytic activity on amorphous PET at 72 °C compared to ICCG, outperforming other PET hydrolases, and exhibited superior degradation activity on real substrates such as cake containers and PET fibers. Conformational analysis revealed the key role of the remodeled β8-α6 loop in substrate binding and overall stability. Collectively, this study explores a promising approach to modifying PET hydrolase and lays a theoretical foundation for advancing bio-circular economy.

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来源期刊

Journal of Hazardous Materials 工程技术-工程：环境

CiteScore

25.40

自引率

5.90%

发文量

3059

审稿时长

58 days

期刊介绍： The Journal of Hazardous Materials serves as a global platform for promoting cutting-edge research in the field of Environmental Science and Engineering. Our publication features a wide range of articles, including full-length research papers, review articles, and perspectives, with the aim of enhancing our understanding of the dangers and risks associated with various materials concerning public health and the environment. It is important to note that the term "environmental contaminants" refers specifically to substances that pose hazardous effects through contamination, while excluding those that do not have such impacts on the environment or human health. Moreover, we emphasize the distinction between wastes and hazardous materials in order to provide further clarity on the scope of the journal. We have a keen interest in exploring specific compounds and microbial agents that have adverse effects on the environment.