单宁酸对小麦醇溶蛋白结构及功能性质的影响
Effects of Tannic Acid on the Structure and Functional Properties of Wheat Gliadin
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摘要: 采用荧光分光光度计、紫外分光光度计(UV-visible)、傅立叶红外光谱仪(FTIR)等探究单宁酸与小麦醇溶蛋白通过非共价与共价方式结合的机理以及蛋白结构与功能特性的改变。结果表明,在非共价作用下,单宁酸对小麦醇溶蛋白的淬灭类型为生成复合物的静态淬灭作用,两者以氢键作用驱动结合,且以1:1摩尔比结合。同时,在碱性环境下,单宁酸与小麦醇溶蛋白上的自由氨基发生席夫碱反应以共价方式形成复合物。此外,不同条件下,复合物的形成会使得小麦醇溶蛋白三级结构发生去折叠现象、二级结构展开。碱性环境下复合物的形成导致小麦醇溶蛋白二级结构展开程度更高。单宁酸的加入使复合物的抗氧化能力提高,且非共价复合物抗氧化活性更高。在热稳定性方面,两者共价结合使小麦醇溶蛋白的热稳定性提升1.03℃,而两者非共价结合使小麦醇溶蛋白热稳定性降低5.39℃。Abstract: The binding mechanism between gliadin and tannic acid was investigated by Fluorescence spectrophotometer,UV-visible spectrophotometer(UV-visible),and Fourier transform infrared(FTIR),and changes on structure and functional properties of wheat gliadin were also investigated. The results showed that the quenching type of wheat gliadin by tannic acid under noncovalent action was the static quenching of the compound,which was driven by hydrogen bond and combined with molar ratio 1:1. At the same time,tannic acid reacted with a free amino group on gliadin to form covalent conjugate under alkaline condition. In addition,the formation of the non-covalent mixture or covalent conjugate both caused the unfolding of gliadin. Furtherly,the formation of covalent conjugate resulted in higher effect on the gliadin secondary structure. The addition of tannic acid made the antioxidant capacity of the complex increase,and the non-covalent mixture exhibited higher antioxidant activity. Finally,gliadin-TA covalent conjugate increased the thermal stability of gliadin by 1.03℃,while the non-covalent mixture reduced the thermal stability of gliadin by 5.39℃.