凝胶色谱法分离纯化棉籽低聚糖和蛋白  

Separation and Purification of Raffinose and Proteins by Gel Permeation Chromatography

作　　者：刘聪 鲍宗必[1] 邢华斌[1] 杨亦文[1] 苏宝根[1] 任其龙[1] 

机构地区：[1]浙江大学生物质化工教育部重点实验室,浙江大学化学工程与生物工程学系,浙江杭州310027

出　　处：《高校化学工程学报》2014年第3期503-509,共7页Journal of Chemical Engineering of Chinese Universities

基　　金：国家"十二五"科技支撑计划项目(2011BAD23B03);国家自然科学基金项目(20936005)

摘　　要：棉籽低聚糖是一种功能性低聚糖,可应用于食品、化妆品、医疗等方面。目前,棉籽低聚糖与蛋白的分离是最大的技术难点之一。作者研究了棉籽低聚糖和蛋白的凝胶色谱法分离。以棉籽糖和牛血清白蛋白混合溶液为原料,采用Sephdex G-10、Sephdex LH-20和Toyopearl HW-40C三种不同凝胶制备柱为固定相,考察了凝胶种类、温度、流动相组成、进样浓度等因素对分离的影响,确定了适宜的工艺操作条件:以去离子水为流动相,HW-40C凝胶为固定相,柱温40°C,上样浓度100 mg?mL-1,进样体积1 mL,流速为1.82 BV?h-1。在该工艺条件下,脱脂棉粕提取液经凝胶色谱分离可得到纯度达89.1%的棉籽糖,收率为64.8%。本方法具有操作简便,分离效率高,产品质量好,为棉籽低聚糖和棉籽蛋白的制备研究提供了一种新的方法。Raffinose is a functional oligosaccharide that can be used in food, cosmetics and organ transplant engineering. Currently, the separation of raffinose and proteins is a problem. Gel permeation chromatography was used to separate raffinose and BSA, and factors such as gel packing type, temperature, mobile phase composition and feed concentration were investigated. An optimized chromatographic condition was using Toyopearl HW-40C as the stationary phase and deionized water as the mobile phase with a flow rate of 1.82 BV·h-1. Moreover, the process was operated at 40＆#176;C with feed concentrations up to 100 mg·mL-1. A raffinose recovery of 64.8%and purity of 89.1%are obtained from raffinose extract under such conditions. This method is simple and effective in removing plant proteins for oligosaccharide purification.

关 键 词：凝胶色谱法 低聚糖 蛋白 HW-40C凝胶 棉籽糖 牛血清白蛋白 

分 类 号：TQ464.1[化学工程—制药化工]