阳离子层析富集生牛乳中乳铁蛋白中试技术的研究

Study on Pilot-Scale Technology of Enrichment of Lactoferrin in Raw Milk by Cation Chromatography

乳铁蛋白是乳汁中一种微量的非血红素铁结合活性糖蛋白,具有抵抗病毒入侵、促进细胞生长、调节肠道内菌群平衡等功能,对人畜健康具有非常重要的作用。其在人乳中的含量是牛乳中含量的10倍以上。因此,多年来我国多种注册的婴儿配方乳粉都含有乳铁蛋白。然而牛乳中乳铁蛋白富集制备复杂,且其结构和活性会受到温度、p H值等多种因素的影响,因此,此类蛋白国内未有生产,一直依赖进口。文章以生牛乳为原料,对乳铁蛋白的中试层析技术进行了系统的研究:脱脂乳在室温下泵入离子交换层析柱中,线性流速为400 cm/h,p H值为7的氯化钠PB缓冲溶液浓度分别为0.4、1.0 mol/L低速洗脱杂蛋白和乳铁蛋白,所制备的乳铁蛋白纯度达到95%以上,收率达75%以上。

Lactoferrin is a trace of non heme iron binding active glycoprotein in milk.It has the functions of resisting virus invasion,promoting cell growth and regulating the balance of intestinal flora.It plays a very important role in human and animal health.Its content in human milk is more than 10 times that in cow’s milk.Therefore,over the years,lactoferrin has been added to a variety of registered infant formula in China.However,the enrichment and preparation of lactoferrin in milk is complex,and its structure and activity are affected by temperature,pH and other factors.Therefore,this kind of protein has not been produced in China and has been dependent on import.In this paper,the pilot scale chromatography technology of lactoferrin was systematically studied with raw milk as raw material:skimmed milk was pumped into ion exchange chromatography column at room temperature,the linear flow rate was 400 cm/h,the concentration of sodium chloride PB buffer solution with pH value of 7 was 0.4 mol/L and 1.0 mol/L,respectively.The purity of lactoferrin was more than 95%and the yield was more than 75%.