Division of Nutritional Sciences, 905 S. Goodwin Avenue, University of Illinois, Urbana 61801, USA.
J Dairy Sci. 2013 Jun;96(6):3488-97. doi: 10.3168/jds.2012-6122. Epub 2013 Mar 30.
Milk fat is encapsulated in a milk fat globule membrane (MFGM) that contains bioactive glycoproteins and glycolipids. The MFGM inhibits infectivity of rotavirus (RV), activity that has been attributed to its glycoprotein and carbohydrate components. However, previous studies of proteins and oligosaccharides in the MGFM have not accounted for all the bioactivity associated with the complete MFGM. The lipid fraction of the MFGM accounts for half of its composition by weight, and we postulate that this fraction should be tested by itself to determine if it plays a role in antiviral activity. Herein, the anti-RV activity of an organic extract of MFGM was tested. Natural and whey buttermilk powders containing bovine MFGM enriched in polar lipids were prepared by microfiltration and supercritical fluid extraction treatment to reduce the triglyceride content of the powders. Lipid fractions were then extracted from the MFGM using both single- and dual-phase extraction methods. Whole MFGM and organic extracts were screened in MA-104 cells for anti-infective activity against a neuraminidase-sensitive rotavirus using a focus-forming unit assay. Dose-dependent inhibition was observed for whole buttermilk and cheese whey MFGM against the rotavirus. In general, buttermilk MFGM exhibited greater RV percentage inhibition than cheese whey MFGM. Organic-soluble anti-RV compounds were identified in bovine MFGM. The most active fraction, isolated by dual-phase extraction and iatrobead chromatography, was free of proteins and highly nonpolar. Further separation of this fraction in a less polar solvent (30:1 chloroform:methanol) resolved at least 5 lipid-containing compounds, which likely contribute to the anti-RV activity associated with bovine MFGM. In summary, lipid components associated with MFGM appear to contribute in large part to the anti-RV activity associated with the bovine MFGM.
乳脂肪被包裹在乳脂肪球膜(MFGM)中,其中含有生物活性糖蛋白和糖脂。MFGM 可抑制轮状病毒(RV)的感染力,这种活性归因于其糖蛋白和碳水化合物成分。然而,以前对 MFGM 中的蛋白质和寡糖的研究并未解释与完整 MFGM 相关的所有生物活性。MFGM 的脂质部分占其重量的一半,我们推测应该单独测试这部分,以确定它是否在抗病毒活性中发挥作用。在此,测试了 MFGM 的有机提取物的抗 RV 活性。通过微滤和超临界流体萃取处理制备了含有富含极性脂质的牛 MFGM 的天然和乳清黄油粉,以降低粉末中的甘油三酯含量。然后使用单相和双相提取方法从 MFGM 中提取脂质部分。使用焦点形成单位测定法,在 MA-104 细胞中筛选全 MFGM 和有机提取物,以针对神经氨酸酶敏感的轮状病毒测定抗感染活性。观察到整个黄油和奶酪乳清 MFGM 对轮状病毒呈剂量依赖性抑制。一般来说,黄油 MFGM 对 RV 的抑制率高于奶酪乳清 MFGM。在牛 MFGM 中鉴定出了可溶于有机的抗 RV 化合物。通过双相提取和 iatrobead 色谱分离的最活跃部分不含蛋白质且高度非极性。在极性较小的溶剂(30:1 氯仿:甲醇)中进一步分离该部分,可分离出至少 5 种含脂化合物,这些化合物可能与与牛 MFGM 相关的抗 RV 活性有关。总之,与 MFGM 相关的脂质成分似乎在很大程度上促成了与牛 MFGM 相关的抗 RV 活性。