Improving the Catalytic Activity and Thermostability of FAST-PETase with a Multifunctional Short Peptide.

IF 4.8 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biomolecules Pub Date : 2025-06-18 DOI:10.3390/biom15060888

Jun Yang, Binyang Deng, Pingan Liao, Siyu Lin, Liqi Zheng, Xing Yang, Fei Wang, Chao Zhai, Lixin Ma

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引用次数: 0

Abstract

Previous reports indicated that self-assembling amphipathic peptide S1v1 (AEAEAHAH)₂ significantly enhances the soluble expression, thermostability, and activity of the target proteins when fused to them. In order to obtain high-efficiency enzymes for the large-scale degradation of polyethylene terephthalate (PET), this multifunctional peptide was fused to the N- and C-terminus of FAST-PETase, a variant of Ideonella sakaiensis PETase (IsPETase), with a PT-linker (TTVTTPQTS) harbored between the target protein and the multifunctional peptide. Consistent with previous reports, S1v1 increased the solubility of FAST-PETase slightly. Moreover, it increased the activity of FAST-PETase dramatically. The amount of terephthalic acid (TPA) and mono(2-hydroxyethyl) terephthalic acid (MHET) released from PET substrate after 24 h of digestion at 50°C by fusion enzymes bearing N- and C-terminal S1v1 tag was approximately 2.9- and 4.6-fold that of FAST-PETase, respectively. Furthermore, the optimal temperature and thermostability of the fusion proteins increased in comparison with FAST-PETase. The present study provides a novel strategy to improve the depolymerization efficiency of FAST-PETase.

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用多功能短肽提高FAST-PETase的催化活性和热稳定性。

先前的报道表明，自组装两亲肽S1v1 (AEAEAHAH)2在与靶蛋白融合时显著提高了其可溶性表达、热稳定性和活性。为了获得大规模降解聚对苯二甲酸乙二醇酯（PET）的高效酶，将该多功能肽融合到sakaiiideonella PETase （IsPETase）的一种变体FAST-PETase的N端和c端，并在靶蛋白和多功能肽之间嵌入一个pt连接子（TTVTTPQTS）。与先前的报道一致，S1v1略微提高了FAST-PETase的溶解度。此外，它还能显著提高FAST-PETase的活性。在50°C条件下，携带N-和C-末端S1v1标签的融合酶酶切24 h后，PET底物中对苯二甲酸（TPA）和单（2-羟乙基）对苯二甲酸（MHET）的释放量分别约为FAST-PETase的2.9倍和4.6倍。此外，与FAST-PETase相比，融合蛋白的最适温度和热稳定性都有所提高。本研究为提高FAST-PETase的解聚效率提供了一种新的策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomolecules Biochemistry, Genetics and Molecular Biology-Molecular Biology

CiteScore

9.40

自引率

3.60%

发文量

1640

审稿时长

18.28 days

期刊介绍： Biomolecules (ISSN 2218-273X) is an international, peer-reviewed open access journal focusing on biogenic substances and their biological functions, structures, interactions with other molecules, and their microenvironment as well as biological systems. Biomolecules publishes reviews, regular research papers and short communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.