大豆球蛋白(11S)和β-伴大豆球蛋白(7S)是大豆蛋白的主要组分,比较11S和7S与多酚相互作用的差异,对揭示多酚对大豆蛋白结构与功能性质的影响机制十分必要。该研究采用紫外-可见光谱、荧光光谱、红外光谱和等温量热滴定法研究11S和7S分别与单宁酸(tannins acid,TA)、表没食子儿茶素没食子酸酯((–)-epigallocatechin-3 gallate,EGCG)和没食子酸(gallic acid,GA)3种不同多酚之间的相互作用。结果表明,3种多酚均能增强11S和7S蛋白的紫外吸收并引起吸收峰偏移,同时改变氨基酸微环境。3种多酚均通过静态猝灭机制与蛋白质形成稳定复合物,显著降低其内源荧光强度,其中TA的猝灭率最高,11S-TA和7S-TA的猝灭率分别为93.9%和97.9%。3种多酚与11S/7S蛋白的相互作用均为自发放热过程,且11S与多酚的结合位点数多于7S,11S与3种多酚的结合位点数均为10,而7S与TA、EGCG、GA的结合位点数分别为8.6、6.7、5。结合力类型上,11S与TA之间以氢键和静电作用为主,与EGCG和GA之间则以疏水作用和范德华力为主,而7S与3种多酚之间均以氢键和静电作用为主。3种多酚诱导蛋白质二级结构改变,其中TA的影响最显著,表现为7S-TA的α-螺旋含量从30.17%降低至9.57%,无规则卷曲从22.07%增加至38.69%,11S-TA的α-螺旋含量从30.23%降低至10.14%,无规则卷曲从23.40%增加至40.87%。此外,3种多酚均能提升11S和7S的抗氧化活性(TA>EGCG>GA),并显著改善其乳化性能。
Soy glycinin (11S) and β-conglycinin (7S) are the main components of soybean protein.Comparing the differences in the interaction between 11S/7S and polyphenols is essential for revealing the mechanism by which polyphenols affect the structural and functional properties of soybean protein.In this study, the interactions between 11S/7S and tannic acid (TA), epigallocatechin gallate (EGCG), and gallic acid (GA) were analyzed by ultraviolet-visible spectroscopy, fluorescence spectroscopy, infrared spectroscopy, and isothermal titration calorimetry.The results showed that all three polyphenols enhanced the ultraviolet absorption of 11S and 7S proteins and caused the shift of absorption peaks, while changing the amino acid microenvironment.Polyphenols formed stable complexes with proteins through static quenching mechanisms, significantly reducing their endogenous fluorescence intensity.Among them, TA exhibited the highest quenching rate, with 11S-TA and 7S-TA having quenching rates of 93.9% and 97.9%, respectively.All interactions were spontaneous exothermic processes.Moreover, the number of binding sites between 11S and polyphenols was greater than that between 7S and polyphenols.Specifically, the number of binding sites for 11S with the three polyphenols was 10 each, whereas the binding site numbers for 7S with TA, EGCG, and GA were 8.6, 6.7, and 5, respectively.In terms of binding force types, 11S mainly interacted with TA through hydrogen bonds and electrostatic interactions, with EGCG/GA through hydrophobic interactions and van der Waals forces, while 7S mainly interacted with all polyphenols through hydrogen bonds and electrostatic interactions.The three polyphenols induced changes in the secondary structure of the proteins, with TA having the most significant impact.Specifically, the α-helix content of 7S-TA decreased from 30.17% to 9.57%, while the random coil increased from 22.07% to 38.69%.Similarly, the α-helix content of 11S-TA decreased from 30.23% to 10.14%, and the random coil increased from 23.40% to 40.87%.In addition, all three polyphenols enhanced the antioxidant activity of 11S and 7S (TA>EGCG>GA), and significantly improved their emulsifying properties.
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