黏红酵母表面黏附蛋白及肠黏液受体的初步研究  

Preliminary research on the adhesion protein of Rhodotorula glutinis and the mucus receptor

作　　者：李正[1,2] 李健[2] 刘淇[2] 王群[2] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,山东青岛266071

出　　处：《水产学报》2008年第6期939-944,共6页Journal of Fisheries of China

基　　金：国家自然科学基金(30700617);科技部科研院所社会公益研究专项基金(2004DIB4J165);公益性农业行业科研专项基金(nyhyzx07-046)

摘　　要：为获得对大菱鲆肠道具有较强黏附能力的益生菌株,了解其黏附机制,采用体外黏液黏附模型及Western-blot技术,对黏红酵母等4株益生菌的大菱鲆肠黏液黏附性能及黏红酵母表面粘附蛋白和肠黏液受体进行了初步研究。结果表明,4株益生菌均可黏附于大菱鲆肠黏液;但各菌株的黏附能力存在较大差异,其中黏红酵母黏附能力最强,鼠李糖乳杆菌次之,假丝酵母和枯草芽孢杆菌相对较低。黏红酵母表面蛋白预先与肠黏液孵育2h后,显著抑制了黏红酵母与肠黏液的结合,其黏附百分率下降了83.6%;但上述处理对其他3株益生菌的黏附没有明显影响。Western-blot显示黏红酵母表面蛋白中分子量为38.5ku和28.6ku的两个蛋白参与对肠黏液的黏附过程,大菱鲆肠黏液中分子量为27.3ku和22.3ku两个蛋白可与黏红酵母表面蛋白特异性结合。糖原染色显示上述4个蛋白均为糖蛋白。In order to select probiotic strains with high adhesion capability to turbot intestinal mucus and understand the adhesion mechanism of Rhodotorula glutinis, the ability of Rhodotorula glutinis, Lactobacillus rharanosus, Candida sp. and Bacillus subtilis adhering to the intestinal mucus of turbot （Scophthatmus maximus ） was investigated in vitro, adhesion protein of Rhodotorula glutinis and mucus receptor were also identified. The results indicate all of the four probiotic strains can adhere to turbot intestinal mucus and significantly better than the BSA treatment. Adhesion percent of R. glutinis to intestinal mucus is the highest among the four probiotic strains. C. ethanolica and B. subtilis adhere to intestinal mucus but weakly. Forthermore, adhesion of R. glutinis to intestinal mucus was quantitatively inhibited by graded concentration of surface protein extracted from R. glutinis. However, the surface protein has no effect on the attachment of C. ethanolica and other two strains to the intestinal mucus. The adhesion protein extracted from the R. glutinis cell surface and intestinal mucus isolated from turbot intestine was visualized by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, native polyacrylamide gel eleetrophoresis and was identified by Western-blotting with horseradish peroxidase-labeled mucus or surface proteins. The results show that 38. 5 ku and 28.6 ku protein of cell surface extraction involved in adherence of R. glutinis to mucus. Two kinds of protein in mucus can be recognized by the surface proteins of R. glutinis, the relative molecular weight is 27.3 ku and 22.3 ku, respectively. The above four kinds of proteins were identified as glycoproteins by PAS methods.

关 键 词：黏红酵母 黏附蛋白 黏液受体 大菱鲆 

分 类 号：Q513[生物学—生物化学]