食品科学 ›› 2019, Vol. 40 ›› Issue (23): 8-13.doi: 10.7506/spkx1002-6630-20181112-122

• 基础研究 • 上一篇    下一篇

VD3与大豆分离蛋白相互作用的多重光谱分析与计算

陈爽,王小丹,李瑞,孙洪蕊,王喜波,江连洲   

  1. (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)
  • 出版日期:2019-12-15 发布日期:2019-12-24
  • 基金资助:
    国家大豆产业技术体系项目(CARS-04-PS28)

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

CHEN Shuang, WANG Xiaodan, LI Rui, SUN Hongrui, WANG Xibo, JIANG Lianzhou   

  1. (1. School of Food Science, Northeast Agricultural University, Harbin 150030, China; 2. Jilin Academy of Agricultural Sciences, Changchun 130000, China)
  • Online:2019-12-15 Published:2019-12-24

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1.245×104(293 K)、1.250×104(298 K)、3.531×104(306 K)L/mol,对应的结合位点数分别为0.973 3、0.992 4和1.094 2;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。

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

Abstract: The interaction between vitamin D3 (VD3) and soy protein isolate (SPI) was studied by multiple spectroscopies (?uorescence, synchronous fluorescence, ultraviolet-visible (UV-Vis), and Fourier infrared spectroscopy). The fluorescence spectroscopy results showed that the intrinsic fluorescence of SPI was statically quenched by VD3. The binding constants (KA) at different temperatures were 1.245 × 104 (293 K), 1.250 × 104 (293 K) and 3.531 × 104 (306 K) L/mol, and the numbers of binding sites (n) were 0.973 3, 0.992 4 and 1.094 2, respectively. The binding distance (r) was 2.92, and the binding promoted fluorescence 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 VD3 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 VD3 could indude secondary structure changes in SPI.

Key words: soy protein isolate, vitamin D3, ?uorescence spectroscopy, synchronous fluorescence, ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy

中图分类号: