Gallego-Lobillo Pablo, Lopez-Rodulfo Ivan M, Martinez Mario M
Centre for Innovative Food (CiFOOD), Department of Food Science, Aarhus University, Agro Food Park 48, Aarhus N 8200, Denmark.
Centre for Innovative Food (CiFOOD), Department of Food Science, Aarhus University, Agro Food Park 48, Aarhus N 8200, Denmark; Food Technology Area, Department of Agricultural Engineering, University of Valladolid, Spain.
Food Res Int. 2025 Jan;199:115346. doi: 10.1016/j.foodres.2024.115346. Epub 2024 Nov 19.
Most polyphenols are glycosylated, affecting their uptake, metabolism, and biological activity. However, the attached sugar must be removed before absorption and functionality can take place. Yet, despite the biological and chemical implications of polyphenol (de-)glycosylation, most in vitro digestion assays omit the utilization of intestinal brush border α- and/or β-glycosidases to study polyphenol bioaccessibility and deglycosylation. This study investigated the effect of rat small intestine extract (RSIE) as an affordable source of mammalian α- and β-glycosidases in different food matrices: matrix-devoid whole apple extract, whole apple, apple juice, and apple pomace. Using the INFOGEST 2.0 model, transepithelial polyphenol absorption, UHPLC-ESI-QTOF-MS/MS, and the inclusion of RSIE at the 15 U*mL maltase activity reported in the human epithelium, the role of RSIE in polyphenol bioaccessibility and deglycosylation was explored. Moreover, the effect of the plant cell wall (PCW) matrix on the role of RSIE was mechanistically investigated by comparing whole apple (or pomace) with their respective extracts. 36 glycosylated polyphenols were identified, including 33 β-O-glycosides and 3 α-O-glycosides. The content of bioaccessible polyphenol β-O-glycosides and α-O-glycosides was significantly lower (p < 0.001) when RSIE was present, which resulted in a concomitant generation of the aglycone forms (phloretin, quercetin, ferulic acid, caffeic acid and p-coumaric acid). However, the concentration of aglycones was much lower than the reduction in the concentration of glycosylated polyphenols, strongly suggesting that polyphenols bind to RSIE. Matrix-devoid whole apple extract, or pomace extract, exhibited higher polyphenol bioaccessibility than whole apple or pomace, likely due to reduced interactions between polyphenols and the food matrix. Importantly, these differences in bioaccessibility diminished with RSIE, suggesting that RSIE α-glycosidases cleaved α-glucans and disrupted the PCW structure.
大多数多酚都进行了糖基化修饰，这会影响它们的吸收、代谢和生物活性。然而，在吸收和发挥功能之前，附着的糖必须被去除。然而，尽管多酚（去）糖基化具有生物学和化学意义，但大多数体外消化试验在研究多酚的生物可及性和去糖基化时都没有利用肠刷状缘α-和/或β-糖苷酶。本研究调查了大鼠小肠提取物（RSIE）作为哺乳动物α-和β-糖苷酶的廉价来源，在不同食物基质中的作用：不含基质的全苹果提取物、全苹果、苹果汁和苹果渣。使用INFOGEST 2.0模型、跨上皮多酚吸收、超高效液相色谱-电喷雾-四极杆飞行时间串联质谱（UHPLC-ESI-QTOF-MS/MS），以及按照人体上皮中报告的15 U·mL麦芽糖酶活性加入RSIE，探讨了RSIE在多酚生物可及性和去糖基化中的作用。此外，通过比较全苹果（或苹果渣）及其各自的提取物，从机制上研究了植物细胞壁（PCW）基质对RSIE作用的影响。鉴定出36种糖基化多酚，包括33种β-O-糖苷和3种α-O-糖苷。当存在RSIE时，生物可及性多酚β-O-糖苷和α-O-糖苷的含量显著降低（p < 0.001），这导致了相应苷元形式（根皮素、槲皮素、阿魏酸、咖啡酸和对香豆酸）的产生。然而，苷元的浓度远低于糖基化多酚浓度的降低，这强烈表明多酚与RSIE结合。不含基质的全苹果提取物或苹果渣提取物表现出比全苹果或苹果渣更高的多酚生物可及性，这可能是由于多酚与食物基质之间的相互作用减少。重要的是，随着RSIE的加入，这些生物可及性的差异减小，这表明RSIE α-糖苷酶切割了α-葡聚糖并破坏了PCW结构。
