摘要
为探究酚酸对β-葡萄糖苷酶活性的影响,通过圆二色谱、分子对接等方法,分析了不同质量浓度(0~0.5 g/L)对羟基苯甲酸(p-hydroxybenzoic acid,PHBA)对β-葡萄糖苷酶的激活机制,重点研究了酶动力学和分子相互作用。结果表明,PHBA显著提高了β-葡萄糖苷酶的活性,且呈浓度依赖性模式。二级结构分析发现,在0.5 g/L的PHBA作用下,与对照组相比,α-螺旋含量显著降低了52.14%(P<0.05),而β-折叠、β-转角和无规则卷曲结构分别增加了90.20%、27.24%和50.56%,表明β-葡萄糖苷酶的活性位点可能位于α-螺旋内。荧光猝灭结果表明,PHBA与β-葡萄糖苷酶可能有1个或者1类结合位点,二者结合是自发的。分子相互作用力显示,氢键、范德华力以及疏水相互作用是稳定PHBA-β-葡萄糖苷酶复合物的主要作用力。分子动力学模拟进一步验证了PHBA-β-葡萄糖苷酶复合物在100 ns轨迹中的结构稳定性(RMSD<0.4 nm),证实了PHBA作为β-葡萄糖苷酶激活剂的潜力,为富含酚酸的果蔬发酵产品中β-葡萄糖苷酶活性提升及功能性成分开发提供了理论依据。
To investigate the effect of phenolic acids on the activity ofβ-glucosidase,this study systematically evaluated the activation mechanism of p-hydroxybenzoic acid(PHBA)at varying concentrations(0~0.5 g/L)through circular dichroism spectroscopy and molecular docking,with a focus on enzyme conformational dynamics and molecular interactions.The results showed that PHBA significantly enhancedβ-glucosidase activity in a concentration-dependent manner.Secondary structural analysis revealed that the content ofα-helix significantly decreased by 52.14%(P<0.05),whileβ-sheet,β-turn,and random coil structures increased by 90.20%,27.24%,and 50.56%,respectively,at 0.5 g/L PHBA compared to the control group.These structural shifts suggestted that the enzyme's active site might reside withinα-helix regions.Fluorescence quenching assays confirmed spontaneous binding between PHBA andβ-glucosidase,with one or one class of binding sites identified.Molecular interaction analyses revealed that hydrogen bonding,van der Waals forces,and hydrophobic interactions predominantly stabilized the PHBA-enzyme complex.Molecular dynamics simulations further validated the structural stability of the PHBA-β-glucosidase complex over 100 ns trajectories(RMSD<0.4 nm),confirming PHBA as a potentβ-glucosidase activator.This study elucidates the molecular basis of PHBA-driven enzyme activation and provides a theoretical foundation for enhancingβ-glucosidase activity in phenolic acid-rich fermented fruit and vegetable products.
作者
李顺峰
陈云莉
兰雪怡
冯路瑶
王安建
王小媛
张丽华
LI Shunfeng;CHEN Yunli;LAN Xueyi;FENG Luyao;WANG Anjian;WANG Xiaoyuan;ZHANG Lihua(Research Center of Agro-products Processing Science and Technology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;College of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001,China;Key Laboratory of Cold Chain Food Processing and Safety Control,Ministry of Education,Zhengzhou University of Light Industry,Zhengzhou 450001,China)
出处
《食品工业科技》
北大核心
2026年第4期89-96,共8页
Science and Technology of Food Industry
基金
河南省重点研发专项项目(231111112500,241111111800)
郑州轻工业大学基础研究基金资助项目(23XJCYJ018)。
关键词
对羟基苯甲酸
Β-葡萄糖苷酶
圆二色谱
荧光猝灭
分子对接
p-hydroxybenzoic acid
β-glucosidase
circular dichroism
fluorescence quenching
molecular docking
