双歧杆菌黏附素的提纯及鉴定(2)

２．２双歧杆菌黏附素的鉴定　将硫酸铵沉淀物经Ｓｕｐｅｒｄｅｘ７５凝胶过滤和Ｑ－Ｓｅｐｈａｒｏｓｅ　ＦＦ离子交换层析后，　得到一单一条带蛋白质，　Ｍｒ　１６　ｋｕ（图３）．

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图　１　　双歧杆菌黏附素在Ｓｕｐｅｒｄｅｘ　７５柱分离色谱图．阴影部分含黏附素活性组分．

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图　２　　双歧杆菌黏附素在Ｑ－Ｓｅｐｈａｒｏｓｅ　ＦＦ柱分离色谱图．第一峰含黏附素活性组分．

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１　　分子质量标准；２　　硫酸铵沉淀物；

３　　Ｓｕｐｅｒｄｅｘ７５凝胶过滤活性组分；

４　　Ｑ－Ｓｅｐｈａｒｏｓｅ　ＦＦ离子交换层析活性组分图　３　　双歧杆菌黏附素ＳＤＳ－ＰＡＧＥ分析．

３　　　讨论

双歧杆菌对肠上皮细胞及其分泌的黏液的黏附能力已作为评定其益生作用的主要指标之一，但有关双歧杆菌黏附机制的研究报道甚少［５－１２］　．黏附是细菌黏附素与宿主细胞相应受体之间的特异性结合过程．革兰阳性细菌细胞壁中含有丰富的磷壁酸，因此对革兰阳性细菌黏附素的研究绝大多数集中在磷壁酸，大量的研究已

双歧杆菌资料

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证实，链球菌及葡萄球菌的黏附素为脂磷壁酸（ＬＴＡ）．传统认为双歧杆菌也是通过其表面的ＬＴＡ介导其黏附，但近年的一些研究表明，双歧杆菌的黏附素可能是一种细胞表面相关的或分泌到培养液上清中的蛋白质［４，１３，１４］　．Ｆｕｊｉｗａｒａ在研究双歧杆菌对大肠杆菌黏附抑制作用时，从双歧杆菌培养上清中纯化出一种Ｍｒ　　５２　ｋｕ的蛋白质，发现其能抑制大肠杆菌对ＧＡ１的黏附，可能也是双歧杆菌的黏附素［１５，１６］　．我们在以往研究的基础上，采用硫酸铵沉淀，　凝胶过滤和离子交换色谱法，　从双歧杆菌耗尽培养液上清中纯化出一种Ｍｒ　　１６　ｋｕ的蛋白质，　经证实为双歧杆菌黏附素．这一结果为进一步研究双歧杆菌黏附的基因调控提供了物质基础．细菌在肠道中黏附与定植是一个非常复杂的过程，　往往有多种因素的参与．黏附不仅发生在细菌与宿主细胞之间；而且还涉及到细菌与黏液层之间的相互作用．另外，　有些细菌的黏附可能需要多种黏附素及其受体的参与．我们首次发现，１６　ｋｕ的蛋白质能介导双歧杆菌对体外培养细胞系Ｌｏｖｏ细胞的黏附．但在体内，　双歧杆菌与肠道上皮细胞间的黏附是否还有其他黏附素的参与以及黏液层在黏附中的作用如何，　尚待进一步的研究．４　　　　　　参考文献

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