| 引用本文: | 李海梅,马莺,崔艳华,汪加琦,李启明,何胜华.牦牛乳中主要过敏原的分离纯化[J].哈尔滨工业大学学报,2012,44(4):80.DOI:10.11918/j.issn.0367-6234.2012.04.017 |
| LI Hai mei,MA Ying,CUI Yan hua,WANG Jia qi,LI Qi ming,HE Sheng hua.Separation and purification of major allergens in yak milk[J].Journal of Harbin Institute of Technology,2012,44(4):80.DOI:10.11918/j.issn.0367-6234.2012.04.017 |
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| 摘要: |
| 为研究牦牛乳蛋白质的过敏性,用化学方法分离牦牛乳中主要的过敏原蛋白质,采用快速蛋白纯化系统(FPLC)结合凝胶层析技术对分离出的蛋白质进行纯化．在牦牛脱脂乳中添加10％醋酸至酪蛋白的等电点pH46,使酪蛋白和乳清蛋白分离．根据酪蛋白在尿素溶液中的溶解度和对钙的敏感性不同,对酪蛋白进行粗分级,得到αs 酪蛋白和β 酪蛋白．应用离子交换色谱和葡聚凝胶sephadex G100进一步纯化粗分级得到的酪蛋白,得到αs casein和β 酪蛋白．以柠檬酸钠为螯合剂、在低pH并有盐存在的条件下从酸乳清中分离出β 乳球蛋白,FPLC纯化后用RP HPLC检测其纯度,得到αs casein和β 酪蛋白纯度分别为8713％和8858％,β 乳球蛋白的纯度为8000％． |
| 关键词: 牦牛乳 酪蛋白分离 β 乳球蛋白分离 快速蛋白纯化色谱 |
| DOI：10.11918/j.issn.0367-6234.2012.04.017 |
| 分类号:TS252 |
| 基金项目:国家自然科学基金资助项目(30871953) |
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| Separation and purification of major allergens in yak milk |
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LI Haimei1, MA Ying1, CUI Yanhua1, WANG Jiaqi2, LI Qiming2, HE Shenghua1
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1.School of Food Science and Engineering, Harbin Institute of Technology, 150090 Harbin, China;2.Research and Development Central of New Hope Dairy Company, 610041 Chengdu, China
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| Abstract: |
| The major allergens in yak milk were obtained by chemical methods, and purified by a fast protein liquid chromatography (FPLC) combined gel chromatography in order to analyze allergenicity of protein in yak milk. Casein and whey were isolated after adjusting isoelectric precipitating of skimmed milk to 46 by adding 10％ acetic acid. Based on differential solubility in urea solutions and differential sensitivity of calcium, αs casein and β casein were separated from whole casein. And then, the crude fractions of αs casein, β casein were further separated using anion exchange FPLC and Sephadex G100 column. β lactoglobulin was separated from acid whey using chelating of citrate sodium at low pH in the presence of salt and purified by FPLC. The results of RP HPLC showed that the purities of αs , β casein, and β lactoglobulin were 8713％, 8858％, and 8000％, respectively. |
| Key words: yak milk casein separation β lactoglobulin separation fast protein liquid chromatography(FPLC) |
