裙带菜褐藻糖胶的提取、纯化及组分研究

论文编号:SP129  论文字数:9839,页数:30

摘要：褐藻糖胶是存在于褐藻中的硫酸酯多糖，具有抗凝血、抗病毒等多种生理活性。本实验采用甲醇对新鲜裙带菜进行预处理，分别用水提法和酸提法提取多糖，采用不同浓度乙醇进行分级沉淀。用蒽酮-硫酸法测定各级沉淀中多糖的含量，结果多糖基本上存在60%乙醇浓度时的沉淀中。提取的粗多糖样品采用CaCl2法进行纯化，得到褐藻糖胶。褐藻糖胶采用Sevag法去除蛋白质后，上DEAE-Sepharose Fast Flow离子交换层析进行分级，用蒽酮硫酸法检测洗脱峰，得到五个峰，对应为F1, F2, F3, F4, F5五个组分。用硫酸钡比浊法测定五个组分的硫酸根含量分别为F1为18.3%；F2,为19.2%；F3为30.1%；F4为38.4%；F5为32.96%。多糖组分三氟乙酸水解后，对水解产物进行糖腈乙酸酯衍生化后，气相色谱分析确定F1主要含有甘露糖和半乳糖；F2主要含有鼠李糖，岩藻糖，木糖，甘露糖，葡萄糖和半乳糖；F3主要含有岩藻糖；F4主要含有岩藻糖和半乳糖；F5主要含有岩藻糖和半乳糖。

关键词：裙带菜；岩藻聚糖；硫酸根；单糖组成

Abstract: Fucoidan is a sulfated polysaccharide from brown seaweed, which has a variety of biological activities, such as anticoagulant activities and antiviral activities. In this paper, Fucoidan was extracted from Undaria pinnatifida pretreated with methanol by acid method and aqueous method, and separated by fractional precipitation with alcohol. The fucoidan was found in the deposition when the consistence of ehthol was 60%. The crude fucoidan was purified by CaCl2 method. Then, the protein in fucoidan was removed by sevag way. DEAE-Sepharose Fast Flow column chromatography was used to isolate purification fucoidan, and F1、F2、F3、F4、F5 were obtained. The content of sulfate and the composition of the properties were analyzed. The content of sulfate was as follow: F1, 18.3%; F2, 19.2%; F3, 30.1%; F4, 38.4%; F5, 32.96%. The monosaccharide composition of the properties was identified with trifluoroacetic acid by the method of aldononitrile acetate precolumn-derivatizaion of Gas chromatography. The composition was as follow: F1, mannose and galactose; F2, rhamnose、fucose、xylose、mannose、glaucose and galactose; F3, fucose; F4, fucose and galactose; F5, fucose and galactose.

Keywords: Undaria pinnatifida; Fucoidan; sulfate; monosaccharide composition

目录
中文摘要 I
英文摘要 II
目录 Ⅲ
1 绪论 1
1.1 裙带菜简介 1
1.2 褐藻糖胶简介 2
1.3  褐藻糖胶的研究方法 2
2 实验部分 5
2.1  材料 5
2.1.1  裙带菜(Undaria pinnatifida) 5
2.1.2  试剂和设备 5
2.2  水提法 7
2.3  酸提法 8
2.4  葡萄糖含量标准曲线的绘制 8
2.4.1  试剂 8
2.4.2  标准曲线的绘制 9
2.4.3  粗多糖含量的测定 9
2.5  粗多糖的纯化 9
2.6  Sevag法除褐藻糖胶中蛋白质 9
2.7  考马思亮蓝法测定蛋白质含量标准曲线的绘制 10
2.7.1  试剂 10
2.7.2  蛋白质含量标准曲线的绘制 10
2.8  褐藻糖胶的层析法分离 10
2.8.1  试剂 10
2.8.2  层析方法 11
2.9  硫酸根含量标准曲线的绘制 11
2.9.1  试剂 11
2.9.2  硫酸根含量标准曲线的绘制 12
2.9.3  换算因子（f）的测定 12
2.9.4  褐藻糖胶中硫酸根含量的测定 12
2.10  气相色谱法测定褐藻糖胶中单糖组成 13
2.10.1  标准单糖衍生化 13
2.10.2  褐藻糖胶水解和衍生化 13
2.10.3  气相色谱条件 13
3  结果与讨论 14
3.1  水提法和酸提法的比较 14
3.2  CaC12法纯化粗多糖 14
3.3  褐藻糖胶除蛋白质的检测 14
3.4  柱层析分离 15
3.5  褐藻糖胶中单糖组成分析 17
3.5.1  标准单糖气相色谱图 17
3.5.2  褐藻糖胶组分气相色谱图 18
4 总结与展望 21
4.1  试验总结 21
4.2  展望 21
致谢 23
参考文献 24