【作者】 李娜；

【导师】 张丽萍；

【作者基本信息】 东北师范大学 ， 生物化学与分子生物学， 2006， 硕士

【摘要】 卡介菌多糖核酸(BCG polysaccharide nucleic acid,BCG-PSN)是我国首创的新型治疗和免疫调节剂。主要用于预防和治疗慢性气管炎、感冒和哮喘等疾病。可特异增强呼吸系统器官的免疫反应,从而增强患者的抗病能力,其主要的治疗机理是通过调节机体内细胞免疫、体液免疫、刺激网状内皮系统,激活单核——巨噬细胞功能,增强自然杀伤细胞功能来增强机体的抗病能力。卡介菌多糖核酸是由卡介菌经热酚法提取多糖和核酸,以灭菌生理氯化钠溶液配置而成的,本文选取卡介菌多糖核酸为研究对象,研究其结构特征,具体内容如下:在原有工艺的基础上,改进卡介菌的菌体培养方法和卡介菌多糖核酸的提取、收集方法,提高了卡介菌多糖和核酸制剂的产量和质量。卡介菌多糖核酸主要含有多糖和核酸两种组分,为明确其有效成分,将卡介菌多糖核酸经DEAE-纤维素阴离子柱层析分离,凝胶柱层析方法纯化,得到一种白色多糖——卡介菌多糖(BCG Polysaccharide BCG-PSA)。经高效液相色谱(HPLC)检验,BCG-PSA的图谱呈单一谱带,表明BCG-PSA为均一组分,分子量约3.1万,气相色谱(GC)分析表明其单糖组成为葡萄糖,属均一多糖类。BCG-PSA结构分析采用了高碘酸氧化、Smith降解、IR分析、GC分析、甲基化分析等方法。甲基化产物经酸水解、还原、乙酰化后用GC-MS联机分析。分析结果表明:BCG-PSA为少分支结构,主要由β型糖苷键构成,其主体结构由Glc组成。其中(1→6)Glc构成主链的核心结构,(1→6)Glc在0-4处有分支,支链部分由(1→4)Glc构成,平均每十个己糖残基有2个分支,末端基为1→Glc。更多还原

【Abstract】 BCG Polysaccharide and Nucleic Acid is mainly used in prevention and treatment ofchronic bronchitis cold and asthma. It can especially strengthen the immune reaction of theorgan of respiratory system, and therefore, it can strengthen the disease-resistant ability ofpatients. It can adjust the function humoral immunity and cellular immunity, stimulatereticular-endothelial system, activate mononuclear phagocyte function, and strengthen thefunction of Natural Killer cell.This preparation is a immunity conditioning agent which is made from polysaccharideand nucleic acid that come from BCG through hot phenol method, with the sterilizationphysiological solution of common salt compound. In this paper, the structure feature of BCGPolysaccharide and Nucleic Acid was reported. .This research was on the base of the old technology. Through improving cultivationmethod of BCG, as well as the extracting and collecting methods of BCG polysaccharide andnucleic acid, we could make the output and quality of them raise.BCG Polysaccharides were isolated from BCG Polysaccharide and Nucleic Acid byDEAE-cellulose column chromatography. The BCG Polysaccharide (BCG-PSA) was purifiedby gel chromatography. HPLC analysis indicated the BCG-PSA was homogeneouspolysaccharide because of its single strait symmetry apex. Its molecular weight is 3.1×104.GC analysis indicted that BCG-PSA was only composed of glucose.By means of periodate oxidation, Smith degradation, IR,GC, the structures of BCG-PSAwas identified. Methylation production were hydrolyzed, deoxidize, then analyzed byGC-MS.BCG-PSA is mainly made up of β configuration glycosidic bond and is a lowlybranched molecule with basic structure composed of (1→6)-linked -Glc residues in the mainchain. Some of (1→6)-linked -Glc residues are substituted at 4-0. On average, there are 2branches among 10 units. The side chain is composed of (1→4)-linked –Glc. Thenonreduced end is Glc.更多还原