云芝子实体多糖和菌丝体多糖的分离纯化研究

【作者】 张松；

【导师】 林瑞超；

【作者基本信息】 北京中医药大学 ， 微生物与生化药学， 2009， 硕士

【摘要】 云芝,学名为彩绒革盖菌Corilus versicolor（L.ex Fr.）Quel,属多孔菌科革盖菌属的一种珍贵的药用真菌,现代植物化学及药理学研究表明云芝主要含蛋白质、脂肪、多糖、多糖肽、氨基酸及无机盐等成分,其主要活性成分是云芝多糖（Polysaccharide）或云芝糖肽（Polysaccharidiopeptide）,云芝多糖具多种生理活性,可抑制肿瘤、提高免疫功能,保肝,抗溃疡等,对正常细胞基本没有毒副作用。临床上用于治疗慢性气管炎、慢性肝炎等疾病,还是一种新型的抗肿瘤免疫调节剂。本研究利用多种色谱技术及现代光谱、波谱技术,对云芝子实体多糖和云芝菌丝体多糖的分子量、组成、结构进行了较为系统的比较:经乙醇分级沉淀、DE-52纤维素柱层析、葡聚糖凝胶柱层析分离纯化,得到4个水溶性多糖组分,分别命名为CVP-a、CVP-b、CVP-c、PSP-a。本文分别采用HPGPC法分析纯度并测定其分子量,通过水解产物的TLC及衍生物的GC-MS确定其单糖组成,并经光谱及波谱数据分析(IR、¹H-NMR、¹³C-NMR等)确定了糖苷键的类型。结果表明,四种多糖均由葡萄糖组成,其中CVP-a、CVP-b、CVP-c分子量分别约为1.5×10⁴、1.2×10⁴、1.1×10⁴,均含β-糖苷键。PSP-a分子量为1.7×10⁴,由α-糖苷键构成。本研究通过对云芝子实体多糖和云芝菌丝体多糖的系统比较,以保留时间为参照,初步建立了云芝多糖分子量测定的方法。经对不同浓度的醇沉淀物质进行考察,确定60%的醇沉部分,通过DE-52纤维素层析处理,高效凝胶色谱分析,已可初步区分不同厂家、不同品种的菌类多糖。此为以菌类多糖为原料及其产品——多糖类药物的质量控制提供了思路与方法。更多还原

【Abstract】 Coriolus versicolor(L.ex Fr.) Quel is a traditional Chinese medicine and edible fungus which is a Polyporaceae fungus belonging to the Basidiomycetes. The modern Phytochemistry and Pharmacology research show that it contains many chemical compositions,such as protein,fat,polysaccharides(CVP), Polysaccharidiopeptide(PSP),dextran,lignin,amino acids and mineral salt,ect. The CVP or PSP are its active components.CVP or PSK with a wide range of physical activity,can inhibit the tumor and improve immune function,protect liver, anti-ulcer,and cause no toxic or side effects to normal cells.It is also a new anti-tumor immunoregulant.This research mainly focused on the molecular,component and structure characterization of Coriolus Versicolor Polysaccharide(CVP) and Coriolus versicolor mycelium polysaccharide(PSP) by means of many kinds of modern chromatograph and spectrum technologies.Four water-soluble homogeneous polysaccharides,named CVP-a,CVP-b,CVP-c and PSP-a were purified from Coriolus Versicolor Polysaccharide(CVP) and Coriolus versicolor mycelium polysaccharide(PSP) through grading precipitation with ethanol,DE-52 cellulose chromatography and gel-filtration chromatofraphy repeatedly.HPGPC was used to analyze their purity and determine their molecular weight(Mw).Thin layer chromatography(TLC),GC-MS,infrared spectrum(IR),and NMR were applied to analyze the configuration of glucose..It was showed by the HPGPC that each peak of the four fractions was a smooth balanced single one.So it was deduced that CVP-a,with Mw is about 1.5×10~4,CVP-b,with Mw is about 1.2×10~4 and CVP-c, with Mw is about 1.1×10~4,were all composed of glucose,meanwhile the configuration of glucose wasβ.And PSP-a,with Mw is about 1.7×10~4,was also a composition of glucose,but the configuration of glucose wasβ.This paper compared the Coriolus Versicolor Polysaccharide(CVP) and Coriolus versicolor mycelium polysaccharide(PSP) systematically.Methods of detecting the average molecular weight were established by means of HPGPC using the index of retention time.Precipitated with 60%ethanol,dealed with DE-52 cellulose chromatography and analyzed by HPGPC,CVP and PSP coming form different manufactories and variaties could be distinguished..The method brought up ideas and ways for quality control of Fungi polysaccharide drugs.更多还原