Amigo-Benavent Miryam, Clemente Alfonso, Caira Simonetta, Stiuso Paola, Ferranti Pasquale, del Castillo M Dolores
Institute of Food Science Research (CSIC-UAM), Madrid, Spain.
Electrophoresis. 2014 Jun;35(11):1582-9. doi: 10.1002/elps.201300527. Epub 2014 Feb 7.
This research investigates how in vitro digestion contributes to the release of antioxidant peptides crypted in soybean β-conglycinin (7S) and its deglycosylated form (D7S). It also investigates the uptake of the bioactive peptides by human intestinal Caco-2 cells using a bicameral system, and their effect on the antioxidant cell defense. Phytochemomics is used as a tool for achieving this goal. The peptides are obtained by mimicking human physiological gastrointestinal digestion conditions. The antioxidant capacity of the peptides is tested by ABTS•(+) radical cation decolorization (2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS)) and oxygen radical absorbance capacity assays. The antioxidant power of the peptides recovered from the basolateral chamber is also evaluated by an analysis of biomarkers of cellular oxidative stress such as cell proliferation, alkaline phosphatase, and secretion of nitric oxide, lipid peroxidation, superoxide dismutase and catalase. Peptides from D7S were more active than those of 7S in the modulation of the cell proliferation, oxidative status and differentiation of Caco-2 cells treated with H2 O2 . Differences in the bioactivity of the peptides of both proteins can be explained by analysis of the structural data obtained by mass spectrophotometry. Our findings support the bioavailability of antioxidant peptides of 7S. The antioxidant properties of 7S soy protein were influenced by events such as glycosylation, digestion, and absorption. Deglycosylation seems to be an innovative strategy for improving the properties of 7S. Deglycosylation might enhance 7S antioxidant power and reduce its immunoreactivity. The combined use of advanced analytical techniques and biochemical analyses (phytochemomics) has been a key part of this study.
本研究调查了体外消化如何促进大豆β-伴大豆球蛋白（7S）及其去糖基化形式（D7S）中隐藏的抗氧化肽的释放。研究还使用双室系统研究了人肠道Caco-2细胞对生物活性肽的摄取及其对抗氧化细胞防御的影响。植物化学组学被用作实现这一目标的工具。通过模拟人体生理胃肠道消化条件获得肽。通过ABTS•(+)自由基阳离子脱色（2,2'-偶氮二（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS））和氧自由基吸收能力测定来测试肽的抗氧化能力。还通过分析细胞氧化应激的生物标志物，如细胞增殖、碱性磷酸酶、一氧化氮分泌、脂质过氧化、超氧化物歧化酶和过氧化氢酶，来评估从基底外侧室回收的肽的抗氧化能力。在调节用H2O2处理的Caco-2细胞的细胞增殖、氧化状态和分化方面，来自D7S的肽比7S的肽更具活性。通过对质谱分析获得的结构数据进行分析，可以解释两种蛋白质肽生物活性的差异。我们的研究结果支持7S抗氧化肽的生物利用度。7S大豆蛋白的抗氧化特性受糖基化、消化和吸收等事件的影响。去糖基化似乎是改善7S特性的一种创新策略。去糖基化可能会增强7S的抗氧化能力并降低其免疫反应性。先进分析技术和生化分析（植物化学组学）的联合使用是本研究的关键部分。
