黄泥螺内脏活性多糖的提取、分离与纯化

Study on extraction,separation and purification technology of polysaccharides from the visceral of bullacta exarata

采用碱提醇沉法结合双酶解法对黄泥螺内脏中活性多糖进行了提取、分离和纯化。单因素试验和正交试验确定了最佳提取工艺条件:提取温度80℃、浸提时间4 h、NaOH浓度为0.5mol/L、乙醇与溶液的体积比为3∶1;最佳纯化方法为:两次醇沉结合双酶解法实现多糖的有效分离,TCA法、Sevag法结合DEAE弱阴离子型纤维素柱。复合多糖提取率为16.22%。利用G100葡聚糖凝胶过滤法对复合多糖进行筛分,成功收集到3种不同分子量的多糖:BEP1、BEP2、BEP3。并测得3种多糖的重均分子量分别为分子量为:MW(BEP1)=62185 Da,MW(BEP2)=46057 u,MW(BEP3)=21747 u;含量BEP132.15%,BEP219.48%,BEP348.37%。醋酸薄膜电泳法检测表明所得3种多糖均为单一多糖。

Polysaccharides from the visceral of Bullacta exarata were extracted,isolated and purified by NaOH extraction-ethanol precipitation-double hydrolysis method.Extraction,isolation and purification were optimized as following： extraction temperature 80 ℃;extraction time 4 h;concentration of NaOH,0.5 mol/L and volume ratio of ethanol to solution 3∶1.The optimum purification procedure was two times alcohol precipitation followed by double hydrolysis,TCA and Sevag for removing protein.Then DEAE column chromatography was used to purify the polysaccharides finally.G100 column chromatography was used to separate the polysaccharide compounds into three kinds of polysaccharides（named as BEP1,BEP2 and BEP3） with different molecular weight,62185 u,46057 u and 21747 u,respectively.Proportions of three polysaccharides were 32.15%,19.48% and 48.37%,respectively.The purities of polysaccharides were verified by cellulose acetate membrane electrophoresis and gel chromatography.