Structure-Function Analysis of a Quinone-Dependent Dehydrogenase Capable of Deoxynivalenol Detoxification.
Structure-Function Analysis of a Quinone-Dependent Dehydrogenase Capable of Deoxynivalenol Detoxification.
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DOI：
10.1021/acs.jafc.2c01083
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发表时间：
2022-05
影响因子：
6.1
通讯作者：
Hua Yang;Ruxue Yan;Yue Li;Zhaoxin Lu;X. Bie;Haizhen Zhao;F. Lu;Meirong Chen
中科院分区：
文献类型：
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作者：
Hua Yang;Ruxue Yan;Yue Li;Zhaoxin Lu;X. Bie;Haizhen Zhao;F. Lu;Meirong Chen
吡咯喹啉醌（PQQ）依赖性脱氢酶DepA通过将C3-OH转化为酮基来解毒脱氧雪腐镰刀菌烯醇（DON）。在此，两个晶体结构的DepA及其与PQQ的复合物被确定，连同生化证据证实的DepA与PQQ和DON的相互作用，并揭示了一个独特的酪氨酸残基重要的底物选择。此外，在DepA活性所必需的活性位点上的四个环被鉴定，其中三个环被PQQ稳定，并且在两种结构中不可见的第四个环被认为对于结合DON是重要的，它们一起构成活性位点的盖子。环的初步工程显示了其对酶改进的潜力。该研究为PQQ依赖性脱氢酶如何具有DON转化功能提供了结构见解，并首次显示了PQQ依赖性脱氢酶活性的盖结构的必要性，为基于结构的设计奠定了基础，以提高催化效率。
The pyrroloquinoline quinone (PQQ)-dependent dehydrogenase DepA detoxifies deoxynivalenol (DON) by converting the C3-OH into a keto group. Herein, two crystal structures of DepA and its complex with PQQ were determined, together with biochemical evidence confirming the interactions of DepA with PQQ and DON and revealing a unique tyrosine residue important for substrate selection. Furthermore, four loops over the active site essential for DepA activity were identified, of which three loops were stabilized by PQQ, and the fourth loop invisible in both structures was considered important for binding DON, together constituting a lid for the active site. Preliminary engineering of the loop showed its potential for enzyme improvement. This study provides structural insights into how a PQQ-dependent dehydrogenase is equipped with the function of DON conversion and for the first time shows the necessity of a lid structure for PQQ-dependent dehydrogenase activity, laying foundation for structure-based design to enhance catalysis efficiency.