VD3与大豆分离蛋白相互作用的多重光谱分析与计算  

Multiple Spectroscopy Analysis and Calculation of the Interaction between Vitamin D3 and Soy Protein Isolate

作　　者：陈爽[1] 王小丹 李瑞 孙洪蕊 王喜波[1] 江连洲[1] CHEN Shuang;WANG Xiaodan;LI Rui;SUN Hongrui;WANG Xibo;JIANG Lianzhou(School of Food Science,Northeast Agricultural University,Harbin 150030,China;Jilin Academy of Agricultural Sciences,Changchun 130000,China)

机构地区：[1]东北农业大学食品学院,黑龙江哈尔滨150030 [2]吉林省农业科学院,吉林长春130000

出　　处：《食品科学》2019年第23期8-13,共6页Food Science

基　　金：国家大豆产业技术体系项目(CARS-04-PS28)

摘　　要：利用多重光谱技术(荧光光谱、同步荧光光谱、紫外-可见光谱、傅里叶变换红外光谱)研究VD3与大豆分离蛋白的相互作用。结果表明:VD3能对大豆分离蛋白的内源荧光进行静态猝灭。VD3与大豆分离蛋白在不同温度下相互作用的表观结合常数分别为1.245×10^4(293 K)、1.250×10^4(298 K)、3.531×10^4(306 K)L/mol,对应的结合位点数分别为0.9733、0.9924和1.0942;结合距离r=2.92,其结合时通过非辐射能力转移而促使蛋白质荧光猝灭。热力学数据分析结果表明:VD3与大豆分离蛋白的反应是自发的吸热过程,其相互作用的主要作用力是静电相互作用和疏水相互作用。同步荧光光谱和紫外-可见光谱结果显示,VD3的添加使大豆分离蛋白构象发生改变,芳香氨基酸残基的微环境由疏水性向亲水性变化。傅里叶变换红外光谱结果表明VD3引起大豆分离蛋白的二级结构发生改变。The interaction between vitamin D 3(VD3)and soy protein isolate(SPI)was studied by multiple spectroscopies(uorescence,synchronous uorescence,ultraviolet-visible(UV-Vis),and Fourier infrared spectroscopy).The uorescence spectroscopy results showed that the intrinsic uorescence of SPI was statically quenched by VD3.The binding constants(KA)at different temperatures were 1.245×10^4(293 K),1.250×10^4(293 K)and 3.531×10^4(306 K)L/mol,and the numbers of binding sites(n)were 0.9733,0.9924 and 1.0942,respectively.The binding distance(r)was 2.92,and the binding promoted uorescence quenching of the protein through non-radiative energy transfer.According to analysis of the thermodynamic data,the reaction of VD3 with SPI was a spontaneous endothermic process,and the interaction was dominated by electrostatic interaction and hydrophobic interaction.The synchronous fluorescence and UV-Vis spectra illustrated that the addition of VD 3 changed the conformation of SPI,and the microenvironment of aromatic amino acid residues changed from hydrophobic to hydrophilic status.Fourier transform infrared spectroscopy revealed that VD 3 could indude secondary structure changes in SPI.

关 键 词：大豆分离蛋白 VD3 荧光光谱 同步荧光光谱 紫外-可见光谱 傅里叶变换红外光谱 

分 类 号：TS201.2[轻工技术与工程—食品科学]