NWIPB OpenIR
牦牛肝蛋白提取分离研究
其他题名Studies on Extraction and Isolation of Proteins of
张耀兮
学位类型硕士
导师李天才
2009-05-27
学位授予单位中国科学院西北高原生物研究所
学位授予地点西北高原生物研究所
关键词牦牛肝 肝蛋白 提取 分离 纯化
摘要以牦牛肝脏为研究对象,进行牦牛肝蛋白提取分离研究。采用碱提法、盐提法、Tris-HCl缓冲液提取法进行了牦牛肝蛋白提取实验,并通过单因素实验和正交实验进行了浓度、料液比、温度、时间等因素对牦牛肝蛋白提取率的影响系列实验;采用DEAE -52和Sephadex G-200层析柱对牦牛肝蛋白进行了分离纯化,并对纯化蛋白进行了活性分析。主要研究内容包括: 1.鲜牦牛肝中蛋白含量为18.90%。 2.实验建立了牦牛肝蛋白最佳提取方法及其提取条件。最佳提取方法为Tris-HCl缓冲液提取法;最佳提取条件为:提取液为Tris-HCl pH8.5,料液比为1:5,温度为0℃,时间为120min。 3.实验建立了牦牛肝蛋白最佳分离纯化条件。牦牛肝蛋白经硫酸铵分级沉淀、脱盐后经过DAEA -52与Sephadex G-200层析柱,初步认为分离得到八个成分均一的肝蛋白,分别为:蛋白Ⅰ、蛋白Ⅱ、蛋白Ⅲ-1、蛋白Ⅲ-2 、蛋白Ⅳ-1、蛋白Ⅳ-2、蛋白Ⅴ-1和蛋白Ⅴ-2。其中蛋白Ⅰ和蛋白Ⅱ经冷冻干燥后呈淡黄色粉末。 4.对上述纯化的八种蛋白进行了比活力分析,其中蛋白Ⅴ-2比活力最高。电泳分析结果显示:蛋白Ⅰ和蛋白Ⅱ均为多亚基蛋白,分子量均在200KDa以上。
其他摘要This paper studied the extraction isolation and activity of proteins by taking the liver of yak (Bos grunniens Linnaeus.) as materials. The single factor test and the orthogonal experiments were used to inspetct the effect of the solution concentration, extraction temperature, the extraction time, and solid-liquid ratio to the extraction rate of proteins from yake livers. DEAE-52 and Sephadex G-200 column chromatography was used to isolate and purify the yak liver proteins. The main research contents include: 1. The percentage of proteins in the fresh yak liver is 18.90% 2. The experiment constructed the optimal extraction method of proteins from yak livers and the optimal extraction conditions. Tris-HCl buffer extraction method is better, and it’s optimal extraction conditions are: Tris-HCl pH8.5, extraction temperature 0℃,extraction time 120 min,material to solvent 1:5. 3. The experiment constructed the separation and purification technology of the yak liver proteins. After fractionation precipitation by (NH)4SO4, desalinization, concentration, the Extracting solution of crude proteins received eight kinds of proteins that were the proteinⅠ、Ⅱ、Ⅲ-1、Ⅲ-2、Ⅳ-1、Ⅳ-2、Ⅴ-1 andⅤ-2 when DAEA-52 and Sephadex G-200 column chromatography were used. The proteinⅠand Ⅱwere as light yellow powder after freeze-drying. 4. Study on the special activity of eight kinds of proteins above indicated that the proteinⅤ-2 had the highest special activity. SDS-PAGE experiments showed that the proteinⅠand Ⅱwere multi-subunit proteins and their molecular weight were more than 200KDa.
页数53
语种中文
文献类型学位论文
条目标识符http://210.75.249.4/handle/363003/3260
专题中国科学院西北高原生物研究所
推荐引用方式
GB/T 7714
张耀兮. 牦牛肝蛋白提取分离研究[D]. 西北高原生物研究所. 中国科学院西北高原生物研究所,2009.
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