以酿酒酵母为研究对象,比较了完整细胞提取法、稀碱缓冲液提取法及溶菌酶和β-葡聚糖酶复合酶法等三种酵母菌细胞壁蛋白提取方法,分析了不同菌龄酵母细胞壁差异性蛋白。结果表明：溶菌酶和β-葡聚糖酶复合酶液水解纯化好细胞壁提取蛋白的方法具有所得胞壁蛋白条带较多,且纯度较高的优点,确定了此方法为提取酵母细胞壁蛋白的最佳提取方法。同时,通过SDS-PAGE电泳分析发现,不同菌龄酵母细胞壁蛋白存在着较大的差异性,并确定了分子质量在36 ku、17 ku和12 ku为不同酵母代数细胞壁的3个主要差异性蛋白,其中36 ku、17 ku处条带蛋白随着菌龄的增加酵母细胞壁蛋白表达量逐渐减少,而12 ku处条带蛋白随着菌龄的增加酵母细胞壁蛋白表达量逐渐增加。%In this paper, Saccharomyces cerevisiae cell wall proteins were taken for the application of whole cell extracted from cell wall proteins, dilute alkaline buffer extracted proteins from purified cell wall and Lysozyme and 6- glucanase enzyme solution hydrolysis of purified cell wall to extracted cell wall proteins all this three extraction meth- ods, at the same time. The differences between different algebra yeast cell wall proteins has been identified and ana- lyzed. The results show that：the method of lysozyme and ~-glucanase enzyme solution hydrolysis of purified cell wall to extract cell wall proteins can obtain more cell wall protein bands and higher purity, therefore this method is deter- minted the best extraction method for yeast cell wall protein extraction. Withal, by SDS-PAGE electrophoresis analysis found different generation of yeast cell wall proteins with a large difference and identified molecular weight of 36 kDa, 17 kDa and 12 kDa are the three main different cell wall proteins for different generation yeasts. Among that, the ex- pression of 36 kDa, 17 kDa protein bands decreased with the age of yeast increasing, whereas, 12 kDa protein band expression increased with the age of yeast.
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