Protein engineering of stable IsPETase for PET plastic degradation by Premuse

文献类型： 外文期刊

第一作者： Meng, Xiangxi

作者： Meng, Xiangxi;Yang, Lixin;Liu, Hanqing;Li, Qingbin;Xu, Guoshun;Zhang, Yan;Guan, Feifei;Zhang, Yuhong;Zhang, Wei;Wu, Ningfeng;Tian, Jian;Li, Qingbin;Zhang, Yan

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关键词： Protein engineering; Poly(ethylene terephthalate) (PET) biodegra-dation; IsPETasc (PETase from Ideonella Sakaiensis)

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：5.162； 五年影响因子：5.137 ）

ISSN： 0141-8130

年卷期： 2021 年 180 卷

页码：

收录情况： SCI

摘要： Poly(ethylene terephthalate) (PET) is used widely by human beings, but is very difficult to degrade. Up to now, the PET degradation effect of PETase from ideonella sakaiensis 201-F6 (IsPETase) variants with low stability and activity was not ideal. In this study, a mutation design tool, Premuse, was developed to integrate the sequence alignment and quantitative selection of the preferred mutations based on natural sequence evolution. Ten single point mutants were selected from 1486 homologous sequences using Premuse, and then two mutations (W159H and F229Y) with improved stability were screened from them. The derived double point mutant, W159H/F229Y, exhibited a strikingly enhanced enzymatic performance. Its T-m and catalytic efficiency values (k(cat)/K-m) respectively increased by 10.4 degrees C and 2.0-fold using p-NPP as the substrate compared with wild type. The degradation activity for amorphous PET was increased by almost 40-fold in comparison with wild type at 40 degrees C in 24 h. Additionally, the variant could catalyze biodegradation of PET bottle preform at a mean rate of 23.4 mg(PET)/h/mg(enz)(yme). This study allowed us to design the mutation more efficiently, and provides a tool for achieving biodegradation of PET pollution under mild natural environments. (C) 2021 Published by Elsevier B.V.

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