【作者】 王凌云；

【导师】 岑颖洲； 李药兰；

【作者基本信息】 暨南大学 ， 分析化学， 2003， 硕士

【摘要】 本文对采自广东汕头南澳海藻养殖场的羊栖菜Sargassum fusiforme进行了粗多糖提取工艺的优化研究。并通过对羊栖菜粗多糖进行分离、纯化和水解获得10个样品，对其中部分样品进行了较为深入的生物活性研究。 以多糖提取率为指标，分别讨论了温度、提取时间、加水量及pH值对羊栖菜粗多糖提取率的影响程度，并根据L₉（3⁴）正交表确定最佳提取工艺条件为温度80℃，加水量50倍，pH值5及时间5h。 采用乙醇、CaCl₂分级沉淀法与DEAE C-52离子交换柱、Sephadex G-100凝胶柱层析相结合，对羊栖菜粗多糖进行了分离纯化，共得到7个样品。通过控制不同的水解条件，获得3个多糖样品。并检测了上述样品的部分理化指标。 采用四甲基偶氮唑盐比色法（MTT法），测定了6个多糖样品对人肝癌细胞的生长抑制率（细胞毒性）。结果显示各样品在体外对肝癌细胞具有一定的生长抑制作用，但其抑制作用较弱。 采用细胞病变（CPE）抑制法和MTT比色法，研究了羊栖菜多糖及其水解样品对单纯疱疹病毒1型（HSV-1）和柯萨奇病毒（CVB₃）的抗病毒活性和对非洲绿猴肾细胞（Vero）的细胞毒性。结果显示，除两个水解样品外，其他样品均表现出了较强的抗病毒活性，且对Vero细胞无明显的毒副作用。研究发现，羊栖菜多糖不仅具有直接杀灭病毒的作用，而且还可进入细胞或吸附在细胞表面，达到抑制或杀伤病毒的效果。初步认为羊栖菜多糖的抗病毒活性与多糖的类型、多糖的水解程度以及硫酸基的含量等因素有关。更多还原

【Abstract】 Sargassum fusiforme were collected from the cultivate base of algae in Nan’ao island, Shantou city, Guangdong province. The processing conditions of extraction of Sargassum fusiforme polysaccharide (SFPS) were optimized. Ten samples were gained by separation, purification and hydrolysis of SFPS and their biological activities were further studied in the paper.The effects of processing conditions on the extraction of SFPS were respectively studied as guiding by the extraction rate of SFPS. By Lg(34) orthogonal experiment design, the optimum processing conditions of extraction were determined. They were as following: temperature of extracting 80 癈, volume of water 50 folds, time of extracting 5 hours and pH 5.Seven samples were isolated from SFPS by using ethanol fractional precipitation, CaCli precipitation, DEAE C-52 chromatographic separation and Sephadex G-100 chromatographic separation. Three samples were obtained by controlling the different conditions of hydrolysis. Some physical and chemical properties of all samples were studied.The cytotoxicity of six samples on HepGi cell was studied by MTT bioassay. The results showed that all samples had some cytotoxicity on HepG2 cell, but the activity was not very potent.The antiviral effects of all samples on HSV-1 and CVB3 and their cytotoxicity on Vero cell were studied by CPE and MTT method. The results showed all samples except two of hydrolyzed polysaccharides had potent antiviral activities. The cytotoxicity of samples on Vero cell was not detected. The further results indicated that polysaccharides not only can killed the above viruses directly but also can restrained them by getting into cells or absorbing on cells. It can be inferred that the antiviral activity of sample is correlated with the kind of polysaccharide, the degree of hydrolysis, the content of sulfated groups.更多还原