目录
1. 绪论 1
1.1 多糖及糖蛋白的来源及作用 1
1.2 微藻的基本知识 1
1.3 微藻多糖及糖蛋白的生物活性 2
1.4 微藻多糖及糖蛋白的提取、分离与纯化 2
1.5 多糖及糖蛋白的性质和结构特征研究 3
1.6 本论文的研究意义及研究内容 4
2. 实验部分 5
2.1 实验仪器与药品 5
2.2 实验用微藻 6
2.3 实验方法 6
3. 结果与讨论 13
3.1 Sephadex 凝胶层析分离纯化 13
3.2 微藻多糖及糖蛋白理化性质的研究 14
3.3 微藻多糖及糖蛋白结构特征的研究 16
4. 结论 19
4.1 结论 19
4.2 存在问题及建议 19
参考文献 20
致谢 22
附录1外文翻译 23
附录2外文原文 29
附录3实验原始数据 33
摘要
海洋微藻资源丰富,因其生存环境的特异性,使细胞内多糖及糖蛋白等多种活性物质具有特殊的生物和生理作用,在药物、功能食品开发、生物制品及环保等方面的研究潜力逐渐引起研究人员的关注。
本文采用螺旋藻和亚心形扁藻为原料,首先采用渗透冲击协同冻结-融化破碎藻细胞;再经过加热和三氯乙酸除蛋白后,通过乙醇沉淀分别获得了浓度在0-50%和50-75%范围的两种级分的粗多糖及糖蛋白。实验测定了多糖及糖蛋白的理化性质,并经过Sephdex G-150葡聚糖凝胶层析、SDS-PAGE凝胶层析和红外光谱分析对其进行了纯化分离和特征鉴定。
实验结果表明,不同微藻及同种微藻的不同级分的多糖和糖蛋白在结构和理化性质上有相似之处,即螺旋藻多糖和糖蛋白、扁藻多糖和糖蛋白的糖链上均含有硫酸基,且均含有少量的分子量范围为分别在40000dalton和60000dalton左右的两种糖蛋白。其不同之处在于各级扁藻多糖和糖蛋白的溶解度、相对粘度均小于相应级分的螺旋藻多糖和糖蛋白,且扁藻多糖和糖蛋白的糖链上还含有氨基,官能团较螺旋藻多糖和糖蛋白更为复杂。
关键词:螺旋藻、扁藻、多糖和糖蛋白、理化性质、结构特征
ABSTRACT
Marine microalgae resource is very abundant and some active materials in cells such as exdopolysaccharide and glycoprotein have many special biological and physiological functions because of the specificity of survival environment, so the research potential on their applications for medicine, function food development, boil
Two kinds of microalgae such as Spirulina paltensis and Platymonas subcordigoramis were taken as raw materials. The cell was firstly broken by penetrated impulse with frozen-thawing, then some free proteins were wiped off after heating and joining trichloroacetic acid to the solution, and then two kinds of the crude polysaccharides and glycoproteins were obtained by precipitating with the concentrations of 0-50% ethanol and 50-75% ethanol, respectively. Their natures of physical and chemistry were firstly determined by means of some methods, and then the separation and purification of the above polysaccharides and glycoproteins were finished by means of Sephdex G-150 polydextran gel and SDS - PAGE gel chromatography, lastly, the functional group characteristic of the above samples were analyzed by infrared spectrum.
The results showed that the polysaccharides and glycoproteins from the above microalgae are similar in the structural character and in the physical and chemical nature, namely there is sulfuric radical in the sugar chain, and there are two kinds of glycoproteins with the molecular weights of about 40000 dalton and about 60000 dalton . There are some differences in the solubility and the relative viscosity of the above samples. The solubility and the relative viscosity of the polysaccharide and glycoprotein from Platymonas subcordigoramis are smaller than those of the polysaccharide and glycoprotein from Spirulina paltensis in the same ethanol concentration, and the functional group of the former is more complicated than that of the latter besides there is animo group in the sugar chain from Platymonas subcordigoramis.
Keywords: Spirulina paltensis; Platymonas subcordigoramis; polysaccharide and glycoprotein; physical and chemical nature;structural character
1.绪论
1.1多糖及糖蛋白的来源及作用
多糖又称多聚糖[1],由单糖聚合而成,是聚合度大于10的极性复杂大分子,其分子量一般为数万甚至达数百万,是所有生命有机体的重要组分,并在控制细胞分裂和分化、调节细胞生长和衰老以及维持生命有机体的正常代谢等方面有重要的作用。目前已发现的活性多糖有几百种,按其来源不同,可分为真菌多糖、高等植物多糖、藻类地衣多糖、动物多糖、细菌多糖五大类。研究表明,通常天然多糖大多与蛋白结合形成糖蛋白,其所具有的生物活性和生理作用不仅取决于蛋白质,更取决于多糖的活性。海洋藻类资源丰富且种类繁多,近年来大量研究表明海藻多糖及糖蛋白具有多种生物活性,诸如抗病毒、抗肿瘤、抗氧化和增强免疫力等,其在医疗和保健方面所具有的重要的潜在应用价值日益显现,因此海藻多糖及糖蛋白的研究越来越受到研究人员的关注[2]。

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