基于响应面法优化凝胶过滤在分离乳铁蛋白中的工艺研究  

Optimization of gel filtration process for lactoferrin separation based on response surface methodology

作　　者：任帅 汤兴楠 朱建星 张万忠 REN Shuai;TANG Xingnan;ZHU Jianxing;ZHANG Wanzhong(College of Pharmacy and Bioengineering,Shenyang University of Chemical Technology,Shenyang 110142)

机构地区：[1]沈阳化工大学制药与生物工程学院,沈阳110142

出　　处：《中国食品添加剂》2023年第2期115-120,共6页China Food Additives

摘　　要：目前我国乳铁蛋白主要依靠进口,产业化发展相对较慢,相对于其他原料,牛初乳具有廉价、易获得等优点。为低成本获得更多乳铁蛋白,本文对凝胶过滤层析纯化方法进行了优化,并设计了响应曲面试验。研究了磷酸盐缓冲液pH、流速、洗脱液浓度三个不同因素对乳铁蛋白含量的影响,通过设计三因素三水平的响应面试验分析优化凝胶过滤层析在乳铁蛋白纯化中的应用,检测方法采用紫外分光光度法,测量所得样品在475 nm处的OD值,计算出相应的质量浓度,得到最终结果。试验表明:最优工艺条件为磷酸盐缓冲液pH 7.48,流速0.20 mL/min,洗脱液浓度0.42 mol/L,预测乳铁蛋白质量浓度为170.567μg/mL。在此条件下进行三次重复试验并取平均值,实际所得乳铁蛋白质量浓度为166.485μg/mL,相对误差为2.393%,与模型基本吻合。At present,lactoferrin in China mainly depends on import,and its industrialization development is relatively slow.Compared with other raw materials,bovine colostrum is cheap and easy to obtain.In order to obtain more lactoferrin at low cost,the gel filtration chromatography purification method was optimized with response surface method.The effects of phosphate buffer pH,flow rate and concentration of eluent,on the content of lactoferrin were studied.The three factors three levels response surface test was designed to optimize gel filtration chromatography in lactoferrin purification process,UV spectrophotometry was used to measure the samples OD value at 475 nm,and the corresponding mass concentration was calculated,to obtain the final result.The results showed that the optimal process conditions were phosphate buffer pH 7.48,flow rate 0.20 mL/min,eluent concentration 0.42 mol/L,and the predicted lactoferrin concentration was 170.567μg/mL.Under these conditions,three repeated experiments were conducted and the average value was taken,and the actual lactoferrin concentration was 166.485μg/mL,with a relative error of 2.393%,which was basically consistent with the model.

关 键 词：牛初乳 乳铁蛋白 响应面分析法 凝胶过滤层析 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程] TS202.3[轻工技术与工程—食品科学与工程]