高效凝胶过滤色谱法测定啤酒中混浊活性蛋白质的研究及应用  

作　　者：孙浩思[1,2] 王莉娜[1,2] 林智平[1,2] 王宏华[1,2] 胡雪莲[1,2] 刘菲[1,2] 

机构地区：[1] 北京燕京啤酒集团公司技术中心,101300 [2] 啤酒酿造技术北京市重点实验室,101300

出　　处：《啤酒科技》2014年第12期22-30,共9页Beer Science and Technology

基　　金：本项目获得北京市优秀人才培养资助

摘　　要：建立了啤酒中混浊活性蛋白质的提取制备方法和基于高效凝胶过滤色谱的蛋白质分析方法,对混浊活性蛋白进行了系统的分子量分布特点及定量分析,并将其应用在了啤酒非生物稳定性及啤酒样品分析领域高效凝胶过滤色谱法解决了混浊活性蛋白分子量及含量难以测定的问题,对啤酒生产蛋白质分子量的控制也具有重要的指导意义结果表明混浊活性蛋白的分布区间为23.4-150.0 kDa、5.7-23.4 kDa、1.0-5.7 kDa和05-1.0kDa啤酒样品中蛋白质含量最高的分子量包括4.8 kDa,2.4 kDa,3.9 kDa和128.8 kDa方法相对标准偏差为0 3％-2.0％,不需混浊活性蛋白提取制备过程中的透析除盐处理,大大简化了操作流程方法简便快速,准确度高,重复性好最后应用该方法对啤酒稳定剂硅胶、酿造单宁和脯氨酸蛋白酶的作用效果进行了分析研究.结果显示硅胶主要去除0.5-1.0 kDa,酿造单宁去除32.0-55.0kDa,而脯氨酸蛋白酶主要去除51.6-150 0kDa部分蛋白质.It was established of haze active protein extraction and preparation methods in beer and protein analysis method based on high gel filtration chromatography, and we made a system analysis of haze active protein for the molecular weight distribution and quantitative, then applied it into the brewing non-biology stabihty study area and analysed some beer samples. The results showed that the distribution of haze active protein is 23.4-1501） kDa, 5.7-23.4 kDa, 1.0-5.7 kDa and 0.5-1.0 kDa Protein content in the highest amount of beer samples including 4.8 kDa, 2.4 kDa, 3.9 kDa and 128.8 kDa High gel filtration chromatography analysis method settled the problem that weight distribution and quantitative of haze active protein hard to definite, using which will have a significant advising meaning for brewing production protein molecular weight distribution control. Methods RSD is 0.3%-2.0%, and simplified the extraction and preparation process. The superiority is simply and fast, accurate and duplicated.At the end, we used method to the beer stabilizer effect analysis of silica gel, brewing tannin and proline protease, Results show that the silica gel mainly the removal of 0.5-1.0 kDa, the brewing tannin removal of 32.0-55.0kDa, and the proline protein removal of 51.6-150.0kDa protein.

关 键 词：高效凝胶过滤色谱法(HPGFC) 定量分析 啤酒 混浊活性蛋白 非生物稳定性 

分 类 号：TS262.5[轻工技术与工程—发酵工程]