强阳离子交换色谱法从牛初乳中分离纯化乳铁蛋白的研究.pdfVIP

48 2007, Vol. 28, No. 07 食品科学 ※基础研究 1,2 2 2 1,2 卢蓉蓉 ，许时婴 ，王 璋 ，杨瑞金 (1.江南大学 食品科学与技术国家重点实验室，江苏 无锡 214122； 2.江南大学食品学院，江苏 无锡 214122) ：本实验研究了离子交换色谱技术大规模纯化乳铁蛋白的色谱条件。选用的SP Sepharose Fast Flow离子交换 剂对乳铁蛋白有很好的吸附选择性，洗脱速度可达到2L/h。以pH值为7.7～8.0，10mmol/L的磷酸盐为起始缓冲 液，通过改变离子强度进行分步洗脱。先采用0.5mol/L的NaCl溶液选择性洗脱分泌型免疫球蛋白A和乳过氧化物 酶组分，再采用 1mol/L 的NaCl 洗脱LF 组分。经 SDS测定，所得乳铁蛋白组分显示为单一区带，相对分 子质量为80400Da。经等点聚焦测定，所得乳铁蛋白的等电点为8.65。中试生产的精制品中乳铁蛋白的纯度和回 收率分别为94.20%和75.45%。 ：乳铁蛋白；分离纯化；强阳离子交换色谱；中试规模 Purification of Lactoferrin from Bovine Colostrum with Strong Cation Exchange Chromatography on Pilot Scale 1,2 2 2 1,2 LU Rong-rong ，XU Shi-ying ，WANG Zhang ，YANG Rui-jin (1.State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China； 2.School of Food Science and Technology, Jiangnan University, Wuxi 214122, China) Abstract ：A stepwise procedure for purification of lactoferrin (LF) extracted from bovine colostrum by ion exchange chromatography on a pilot scale was investigated. SP Sepharose Fast Flow (SP Sepharose FF) of excellent absorption speciality for LF, was chosen as the ion exchanger with elution rate of 2 L/h. 0.5 mol/L NaCl aqueous solution was used to elute the secretory immunoglobulin A and lactoperoxidase. Then, lactoferrin was eluted with 1.0 mol/L NaCl aqueous buffer. Lactoferrin fraction is shown as a single band in SDSwith molecular weight of 80400 Da. The isoelectric point of lactoferrin is 8.65 determined