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新型玉米赤霉烯酮内酯水解酶 ZHRnZ 的特性及其热稳定性改造。

Characteristics of a Novel Zearalenone Lactone Hydrolase ZHRnZ and Its Thermostability Modification.

机构信息

State Key Laboratory of Animal Nutrition and Feeding, Poultry Nutrition and Feed Technology Innovation Team, College of Animal Science and Technology, China Agricultural University, No. 2. West Road Yuanmingyuan, Beijing 100193, China.

Eyasclub, College of Animal Science and Technology, China Agricultural University, No. 2. West Road Yuanmingyuan, Beijing 100193, China.

出版信息

Int J Mol Sci. 2024 Sep 6;25(17):9665. doi: 10.3390/ijms25179665.

Abstract

Zearalenone (ZEN) is a toxic secondary metabolite produced by the fungi, which widely contaminates grains, food, and feed, causing health hazards for humans and animals. Therefore, it is essential to find effective ZEN detoxification methods. Enzymatic degradation of ZEN is believed to be an eco-friendly detoxification strategy, specifically thermostable ZEN degradation enzymes are needed in the food and feed industry. In this study, a novel ZEN lactone hydrolase ZHRnZ from was discovered using bioinformatic and molecular docking technology. The recombinant ZHRnZ showed the best activity at pH 9.0 and 45 °C with more than 90% degradation for ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL) and α-zearalanol (α-ZAL) after incubation for 15 min. We obtained 10 mutants with improved thermostability by single point mutation technology. Among them, mutants E122Q and E122R showed the best performance, which retained more than 30% of their initial activity at 50 °C for 2 min, and approximately 10% of their initial activity at 60 °C for 1 min. The enzymatic kinetic study showed that the catalytic efficiency of E122R was 1.3 times higher than that of the wild-type (WT). Comprehensive consideration suggests that mutant E122R is a promising hydrolase to detoxify ZEN in food and feed.

摘要

玉米赤霉烯酮(ZEN)是一种由真菌产生的有毒次生代谢物,广泛污染谷物、食品和饲料,对人类和动物的健康构成威胁。因此,寻找有效的 ZEN 解毒方法至关重要。酶降解 ZEN 被认为是一种环保的解毒策略,特别是在食品和饲料工业中需要耐热的 ZEN 降解酶。在本研究中,使用生物信息学和分子对接技术发现了一种来自 的新型 ZEN 内酯水解酶 ZHRnZ。重组 ZHRnZ 在 pH 值为 9.0 和 45°C 时表现出最佳活性,对 ZEN、α-玉米赤霉烯醇(α-ZOL)、β-玉米赤霉烯醇(β-ZOL)和 α-玉米赤霉烯醇(α-ZAL)的降解率超过 90%,孵育 15 分钟后。我们通过单点突变技术获得了 10 个具有提高热稳定性的突变体。其中,突变体 E122Q 和 E122R 表现出最佳性能,在 50°C 下保持超过 30%的初始活性 2 分钟,在 60°C 下保持约 10%的初始活性 1 分钟。酶动力学研究表明,E122R 的催化效率比野生型(WT)高 1.3 倍。综合考虑,突变体 E122R 是一种很有前途的水解酶,可以用于食品和饲料中 ZEN 的解毒。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e7ef/11395237/528823dd8baa/ijms-25-09665-g001.jpg

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