Rahman Md Morshedur, Kim Woan-Sub, Ito Toshiaki, Kumura Haruto, Shimazaki Kei-ichi
Laboratory of Dairy Food Science, Research Faculty of Agriculture, Hokkaido University, North 9, West 9, Hokkaido, Sapporo 060-8589, Japan.
Anaerobe. 2009 Aug;15(4):133-7. doi: 10.1016/j.anaerobe.2009.01.003.
Lactoferrin, a major whey protein of human milk, is considered as growth promoter for bifidobacteria, the predominant microorganisms of human intestine. In the present study, in vitro growth promotion and cell binding ability of bovine lactoferrin to several strains of Bifidobacterium longum have been demonstrated. A dose-dependent as well as strain-dependent growth promotion effect by lactoferrin was observed. Cell binding ability of lactoferrin was inspected under an inverted confocal laser scanning microscope by incubation bacterial cells with biotinylated bovine lactoferrin and FITC-conjugated avidin. Fluorescence staining showed bovine lactoferrin binding to all tested strains. A lactoferrin-binding protein with a molecular weight of approximately 67 kDa was also detected in the extracted membrane and cytosolic fraction of each B. longum strain by far-Western blot technique using biotinylated lactoferrin and horseradish peroxidase-conjugated streptavidin. Based on these results, we suggest that existence of lactoferrin-binding protein could be a common characteristic in bifidobacteria. It can also be hypothesized that lactoferrin-binding protein in bifidobacteria is not only involved in growth stimulation mechanism but also could play different roles.
乳铁蛋白是母乳中的一种主要乳清蛋白,被认为是双歧杆菌(人体肠道中的主要微生物)的生长促进剂。在本研究中,已证明牛乳铁蛋白对几株长双歧杆菌具有体外生长促进作用和细胞结合能力。观察到乳铁蛋白具有剂量依赖性和菌株依赖性的生长促进作用。通过将细菌细胞与生物素化牛乳铁蛋白和异硫氰酸荧光素(FITC)偶联抗生物素蛋白一起孵育,在倒置共聚焦激光扫描显微镜下检测乳铁蛋白的细胞结合能力。荧光染色显示牛乳铁蛋白与所有测试菌株结合。使用生物素化乳铁蛋白和辣根过氧化物酶偶联链霉抗生物素蛋白的远缘Western印迹技术,在每种长双歧杆菌菌株的提取膜和胞质组分中还检测到一种分子量约为67 kDa的乳铁蛋白结合蛋白。基于这些结果,我们认为乳铁蛋白结合蛋白的存在可能是双歧杆菌的一个共同特征。还可以推测,双歧杆菌中的乳铁蛋白结合蛋白不仅参与生长刺激机制,而且可能发挥不同作用。