表没食子儿茶素没食子酸酯对乙醇脱氢酶的激活作用:多光谱法和分子对接法研究  

Activation of Epigallocatechin Gallate on Alcohol Dehydrogenase:Multispectroscopy and Molecular Docking Methods

作　　者：张潇丹 刘丽莉[1] 于影 程伟伟 徐宝成[1] 贺家亮[1] 陈树兴[1,2] ZHANG Xiao-dan;LIU Li-li;YU Ying;CHENG Wei-wei;XU Bao-cheng;HE Jia-liang;CHEN Shu-xing(College of Food and Bioengineering,Henan University of Science and Technology,National Experimental Teaching Demonstration Center for Food Processing and Security,Henan International Joint Laboratory of Food Processing and Quality and Safety Control,Research and Utilization of Functional Food Resources Science and Technology Innovation Team of Henan Provincial Department of Education,Luoyang 471023,China;Food Laboratory of Zhongyuan,Luohe 462300,China)

机构地区：[1]河南科技大学食品与生物工程学院,食品加工与安全国家级教学示范中心,食品加工与质量安全控制河南省国际联合实验室,功能食品资源研究与利用河南省教育厅科技创新团队,河南洛阳471023 [2]中原食品实验室,河南漯河462300

出　　处：《光谱学与光谱分析》2023年第11期3622-3628,共7页Spectroscopy and Spectral Analysis

基　　金：2021年洛阳市社会发展专项(2101021A);国家重点研发计划项目(2022YFF1101600);河南省重大科技专项(221100110500);国家自然科学基金项目(U1704114)资助。

摘　　要：乙醇脱氢酶(ADH)在酒精代谢过程中起着关键作用,通过激活ADH活性可以促进人体对乙醇的吸收,进而解酒保肝。对ADH与表没食子儿茶素没食子酸酯(EGCG)的相互作用进行探究,采用紫外可见光谱、荧光光谱、傅里叶变换红外光谱等多光谱法和分子对接法研究了ADH与EGCG的结合机理,采用差示扫描量热仪测定ADH和EGCG-ADH复合物的热变性温度,进而分析二者的热稳定性变化,并通过扫描电镜表征EGCG-ADH复合物的形貌和结构。研究表明,EGCG激活了ADH的催化活性,激活率为33.33%。EGCG作用于ADH引起其微环境变化和二级结构变化,形成了结合位点数接近于1的复合物,范德华力和氢键对其稳定性起着重要作用;相较于ADH,EGCG-ADH的二级结构中α-螺旋含量降低而β-折叠含量升高;分子对接结果进一步证实了EGCG苯环羟基与周围氨基酸之间的氢键有利于保持配合物的稳定性,而EGCG和ADH之间存在的范德华力和正烷基是ADH活性被激活的主要原因。结果证明EGCG通过与ADH结合,激活ADH的催化活性,可为制备更加安全高效的解酒剂替代品提供理论指导。Alcohol dehydrogenase(ADH)plays a key role in the pathway of alcohol metabolism.By activating ADH activity,the absorption of alcohol can be promoted to relieve alcoholism and protect the liver.This paper studied the interaction between ADH and epigallocatechin gallate(EGCG).The binding mechanism of ADH and EGCG was investigated by UV-Vis spectrum,fluorescence spectrum,Fourier infrared spectrum and molecular docking method.The thermal denaturation temperatures of ADH and EGCG-ADH complex were measured by differential scanning calorimeter,and then the thermal stability changes of the complex EGCG were analyzed.EGCG was characterized by scanning electron microscope.The results showed that EGCG activated the catalytic activity of ADH,and the activation rate was 33.33%.The effect of EGCG on ADH caused its microenvironment and secondary structure changes,forming a complex with anumber of binding sites close to 1,and van der waals force and hydrogen bond played an important role in its stability.Compared with ADH,theα-helix content in the secondary structure of the complex decreased,and theβ-sheet content increased.In addition,the molecular docking results further confirmed that the hydrogen bond between the hydroxyl group of the EGCG benzene ring and the surrounding amino acids is beneficial to maintain the stability of the complex.In addition,the van der waals force and n-alkyl between EGCG and ADH are the main reasons for the activation of ADH activity.The above results proved that EGCG can activate the catalytic activity of ADH by combining with ADH,which can provide theoretical guidance for the preparation of safer and more efficient alternatives to hangovers.

关 键 词：乙醇脱氢酶 表没食子儿茶素没食子酸酯 相互作用 光谱法 分子对接 

分 类 号：TS251.95[轻工技术与工程—农产品加工及贮藏工程]