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Structural characterization and functional evaluation of lactoferrin-polyphenol conjugates formed by free-radical graft copolymerization

机译:自由基接枝共聚形成的乳铁蛋白-多酚共轭物的结构表征和功能评价

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Covalent modifications of lactoferrin (LF) with epigallocatechin gallate (EGCG), chlorogenic acid (CA) and gallic acid (GA) were performed by adopting a free-radical grafting procedure in aqueous media. The resulting LF-polyphenol conjugates were characterized in terms of structural and functional properties. Results showed that the covalent binding amount into the LF molecule of EGCG, CA and GA was 68, 58 and 17 nmol mg(-1), respectively. Covalent insertion of polyphenols into the LF molecule was verified by SDS-PAGE and MALDI-TOF-MS analysis, and in particular the molecular weight was increased from 84 011 Da (LF) to 85 906 Da (LF-CA conjugate). The circular dichroism and Fourier transform infrared spectroscopy analyses revealed that the content of alpha-helix increased and the contents of the remaining structures decreased, while the differential scanning calorimetry data indicated that the thermal stability of LF-polyphenol conjugates was enhanced after the modification. In addition, the antioxidant activity of LF-polyphenol conjugates was 0.23- to 2.10-fold (ABTS(center dot+) scavenging assay), and 0.04- to 2.19-fold (reducing power assay) higher than that of the control LF. Moreover, the covalent modification obviously changed the solubility and emulsifying properties of LF. The emulsifying properties of the LF-CA conjugate were better than those of the LF-EGCG and LF-GA conjugates.
机译:乳铁蛋白(LF)与表没食子儿茶素没食子酸酯(EGCG),绿原酸(CA)和没食子酸(GA)的共价修饰是通过在水性介质中采用自由基接枝方法进行的。根据结构和功能性质表征了所得的LF-多酚共轭物。结果表明,EGCG,CA和GA与LF分子的共价结合量分别为68、58和17 nmol mg(-1)。通过SDS-PAGE和MALDI-TOF-MS分析证实多酚共价插入LF分子中,尤其是分子量从84 011 Da(LF)增加到85 906 Da(LF-CA共轭物)。圆二色性和傅立叶变换红外光谱分析表明,α-螺旋的含量增加而其余结构的含量降低,而差示扫描量热法数据表明,改性后LF-多酚共轭物的热稳定性增强。此外,LF-多酚共轭物的抗氧化活性为0.23至2.10倍(ABTS(中心点+)清除测定),比对照LF高0.04至2.19倍(降低功率测定)。而且,共价改性明显改变了LF的溶解性和乳化性能。 LF-CA缀合物的乳化性能优于LF-EGCG和LF-GA缀合物的乳化性能。

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