Enhancing the thermal stability and activity of zearalenone lactone hydrolase to promote zearalenone degradation via semi-rational design

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology Pub Date : 2024-08-22 DOI:10.1016/j.enzmictec.2024.110499

Xue Jiang , Sana Tehreem , Kashif Rahim , Meixing Wang , Pan Wu , Guimin Zhang

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引用次数: 0

Abstract

Zearalenone (ZEN) is a fungal toxin produced by Fusarium exospore, which poses a significant threat to both animal and human health due to its reproductive toxicity. Removing ZEN through ZEN lactonase is currently the most effective method reported, however, all published ZEN lactonases suffer from the poor thermal stability, losing almost all activity after 10 min of treatment at 55℃. In this study, we heterologously expressed ZHD11A from Phialophora macrospora and engineered it via semi-rational design. A mutant I160Y-G242S that can retain about 40 % residual activity at 55℃ for 10 min was obtained, which is the most heat-tolerant ZEN hydrolase reported to date. Moreover, the specific activity of the I160Y-G242S was also elevated 2-fold compared to ZHD11A from 220 U/mg to 450 U/mg, which is one of the most active ZEN lactonses reported. Dynamics analysis revealed that the decreased flexibility of the main-chain carbons contributes to increased thermal stability and the improved substrate binding affinity and catalytic turnover contribute to enhanced activity of variant I160Y-G242S. In all, the mutant I160Y-G242S is an excellent candidate for the industrial application of ZEN degradation.

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通过半合理设计提高玉米赤霉烯酮内酯水解酶的热稳定性和活性，促进玉米赤霉烯酮降解。

玉米赤霉烯酮（Zearalenone，ZEN）是一种由镰刀菌外生孢子产生的真菌毒素，由于具有生殖毒性，对动物和人类健康构成严重威胁。目前，通过 ZEN 内酯酶去除 ZEN 是最有效的方法，但已发表的所有 ZEN 内酯酶都存在热稳定性差的问题，在 55℃ 下处理 10 分钟后几乎丧失所有活性。在本研究中，我们异源表达了大孢子藻中的 ZHD11A，并通过半合理设计对其进行了工程化。我们得到了一个突变体 I160Y-G242S，它在 55℃ 处理 10 分钟后仍能保持约 40% 的残余活性，是迄今为止报道的最耐热的 ZEN 水解酶。此外，与 ZHD11A 相比，I160Y-G242S 的比活性也提高了 2 倍，从 220 U/mg 提高到 450 U/mg ，是目前报道的活性最高的 ZEN 内切酶之一。动力学分析表明，主链碳的灵活性降低有助于提高热稳定性，而底物结合亲和力和催化周转率的提高则有助于增强变体 I160Y-G242S 的活性。总之，突变体 I160Y-G242S 是 ZEN 降解工业应用的绝佳候选物。

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

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.