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揭示脱氧雪腐镰刀菌烯醇氧化酶 DepA 的广泛底物特异性及其在脱毒单端孢霉烯族真菌毒素中的作用。

Unveiling the Broad Substrate Specificity of Deoxynivalenol Oxidation Enzyme DepA and Its Role in Detoxifying Trichothecene Mycotoxins.

机构信息

Guelph Research and Development Centre, Agriculture and Agri-Food Canada, Guelph, ON N1G 5C9, Canada.

出版信息

Toxins (Basel). 2024 Mar 5;16(3):136. doi: 10.3390/toxins16030136.

DOI:10.3390/toxins16030136
PMID:38535802
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10975547/
Abstract

DepA, a pyrroloquinoline quinone (PQQ)-dependent enzyme isolated from mutans 17-2-E-8, exhibits versatility in oxidizing deoxynivalenol (DON) and its derivatives. This study explored DepA's substrate specificity and enzyme kinetics, focusing on DON and 15-acetyl-DON. Besides efficiently oxidizing DON, DepA also transforms 15-acetyl-DON into 15-acetyl-3-keto-DON, as identified via LC-MS/MS and NMR analysis. The kinetic parameters, including the maximum reaction rate, turnover number, and catalytic efficiency, were thoroughly evaluated. DepA-PQQ complex docking was deployed to rationalize the substrate specificity of DepA. This study further delves into the reduced toxicity of the transformation products, as demonstrated via enzyme homology modeling and in silico docking analysis with yeast 80S ribosomes, indicating a potential decrease in toxicity due to lower binding affinity. Utilizing the response surface methodology and central composite rotational design, mathematical models were developed to elucidate the relationship between the enzyme and cofactor concentrations, guiding the future development of detoxification systems for liquid feeds and grain processing. This comprehensive analysis underscores DepA's potential for use in mycotoxin detoxification, offering insights for future applications.

摘要

DepA 是从 mutans 17-2-E-8 中分离出来的一种吡咯喹啉醌 (PQQ) 依赖性酶，它在氧化脱氧雪腐镰刀菌烯醇 (DON) 及其衍生物方面表现出多功能性。本研究探讨了 DepA 的底物特异性和酶动力学特性，重点研究了 DON 和 15-乙酰-DON。DepA 除了能够有效地氧化 DON 外，还能将 15-乙酰-DON 转化为 15-乙酰-3-酮-DON，这是通过 LC-MS/MS 和 NMR 分析确定的。全面评估了动力学参数，包括最大反应速率、 turnover 数和催化效率。DepA-PQQ 复合物对接被用于合理化 DepA 的底物特异性。本研究还进一步探讨了转化产物毒性降低的情况，通过酶同源建模和与酵母 80S 核糖体的计算机对接分析表明，由于结合亲和力降低，毒性可能降低。利用响应面法和中心复合旋转设计，建立了数学模型来阐明酶和辅因子浓度之间的关系，为液体饲料和谷物加工的解毒系统的未来发展提供指导。这项全面的分析强调了 DepA 在真菌毒素解毒方面的应用潜力，为未来的应用提供了思路。

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本文引用的文献

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J Agric Food Chem. 2022 Jun 8;70(22):6764-6774. doi: 10.1021/acs.jafc.2c01083. Epub 2022 May 25.
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