灰树花多糖的提取、化学修饰及活性测定-生物化学与分子生物学专业论文.docxVIP

摘要灰树花多糖具有多种药理活性如增强免疫功能抗肿瘤调节血糖 降低血压等本论文探讨了灰树花多糖中最为著名的一种 D 组分的提取 化学修饰及免疫活性从灰树花Maitake 子实体中提取分离得到子实体多糖 D-组分进 行了理化性质分析考察了提取中的几个关键因素对 D-组分得率的影响 包括加水倍数浸提时间醇沉浓度浓缩倍数根据正交实验结果可知, 加水倍数和抽提时间对灰树花 D 组分得率有显著性影响浓缩比重和醇沉 浓度在所选的水平上对 D 组分得率影响较弱对于工业化生产而言考虑 到生产成本应选择尽可能大的浓缩比重和较小的醇沉浓度由此确定灰 树花子实体多糖 D 组分提取工艺最佳参数为加水倍数 25 倍抽提时间 为 6 个小时纯沉浓度 60%,浓缩比重 1.050对灰树花 D 组分进行硫酸酯化修饰得到了几种不同取代度的硫酸化D 组分并研究了硫酸化反应的条件对取代度的影响最后考察了灰树花 D 组分和其硫酸化衍生物对小鼠的免疫调节作用 研究了灰树花多糖 D 组分及硫酸化灰树花多糖 D 组分对小鼠的免疫器官重 量腹腔巨噬细胞吞噬功能血清溶血素形成及淋巴细胞转化率的影响 结果表明在 10mg/kg 剂量下灰树花多糖 D 组分及硫酸化灰树花多糖 D 组分具有免疫增强作用并且在合适的取代度下硫酸化灰树花多糖 D 组分 比灰树花多糖 D 组分表现了更强的免疫调节作用关键词灰树花多糖,D-组分, 提取, 化学修饰, 免疫功能ABSTRACTThe polysaccharides of Grifola Frondosa have many kinds of pharmacological effects, for example, improving immunological function; antitumor; anti-diabetic activity and blood pressure lowering activity etc. In this article, the extraction, chemical modification and immunological activity of Grifola Frondosa D fraction was discussed.D-fraction was isolated from the fruit bodies of Grifola frondosa. Its physical and chemical characters were analyzed. Several key factors (quantity of water added, time of heating, extent of concentration, concentration of EtOH for precipitation) were studied to test their effects on the output of D-fraction. Based on the results of crossed test, the quantity of water and time of heating had a notable influence on the output of D-fraction. The extent of concentration and concentration of EtOH had a minor influence on the output of D-fraction. Considering industrial production, it’ s better to choose the higher extent of concentration and lower concentration of EtOH. So the optimum process for isolation of D-fraction of Grifola Frondosa is 25 times of water added, 6hrs of heating, 60% EtOH for precipitation and 1.05 for density after concentration.Then the D-fraction was sulfated. Sulfated derivatives with different degrees of substitution were obtained. The condition of sulfation was investigated.At last, The immunological function