凝胶过滤色谱纯化乳清蛋白降胆固醇肽的研究  

Study of purification of whey protein hypocholesterolemic peptides by gel filtration chromatography

作　　者：马丽媛[1,2] 李晓东[1,2,3] 庄建鹏 尚尔坤 任红晶[1,2] 李明浩[1,2] 刘璐[1,2] 毕伟伟[1,2] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030 [2]东北农业大学食品学院,哈尔滨150030 [3]东北农业大学食品安全与营养协同创新中心,哈尔滨150030

出　　处：《中国乳品工业》2014年第5期17-20,共4页China Dairy Industry

基　　金：国家科技部"十二五"科技支撑计划项目(2011BAD09B01);2014年度黑龙江省教育厅科学技术研究项目;哈尔滨市科技创新人才专项留学回国人员基金(2012RFLXN033)

摘　　要：从乳清蛋白水解物中纯化降胆固醇活性肽,乳清蛋白经胰蛋白酶水解得到的水解物经超滤粗分离,采用凝胶过滤色谱分离纯化,并对其洗脱流动相种类、pH值和洗脱液浓度进行优化筛选,采用体外检测方法筛选出最佳洗脱条件下的具有胆固醇胶束溶解度抑制活性最高的组分,并用已知分子量的标准蛋白质凝胶层析绘制校正曲线,建立分子量的对数与出峰时间的关系。结果表明,优化的最佳洗脱液为磷酸钠溶液,研究得到了凝胶过滤色谱的校正曲线,在洗脱时间为71 min时得到的组分降胆固醇活性最高,为57.48%,经计算其分子量在2 000～4 700 u范围内。This study was carried out to purify hypocholesterolemic peptides from whey protein hydrolysates. Whey protein was hydrolyzed by trypsin, and then concentrated by ultrafiltration. Hydrolysates were purified with Gel Filtration Chromatography. The elution mobile phase,pH value and the concentration of mobile phase was selected for optimization. Detection methods in vitro was carried out to obtain the faction with the highest inhibition activity of cholesterol micellar solubility. The calibration curve between logarithm of protein molecular weights and appearance time was investigated. The results showed that the optimal eluent was sodium phosphate solution, the calibration curve of gel filtration chromatography was obtained, and the fraction of eluted at time 71 min had the highest hypocholesterolemic activity which was57.48%, molecular weight distribution as 2 000～4 700 u.

关 键 词：乳清蛋白 降胆固醇肽 分离纯化 活性 

分 类 号：Q93-33[生物学—微生物学]