灵芝孢子粉多糖的分离纯化、结构表征及免疫活性初探

Purification,Structural Characterigation and Immunological Activity of Polysaccharides from the Spores of Ganoderma lucidum

采用水浴回流法提取灵芝孢子粉粗多糖,再依次使用DEAE Sepharose Fast Flow离子交换柱和Sepharose CL-6B凝胶色谱柱分离纯化得到2种多糖组分GLP1a和GLP1b,HPSEC法测得2种组分均呈单一峰,重均分子量分别为1.12×106和1.21×105。HPLC、红外光谱和核磁共振分析的结果表明,GLP1a和GLP1b主要由葡萄糖组成,主链都是以β-(1,6)糖苷键连接,GLP1b在部分1→6连接的葡萄糖的3位和4位有分支。MTT实验表明,2个多糖级分均具有一定的免疫活性,而且都存在明显的量效关系,GLP1b的免疫活性明显高于GLP1a。

The crude polysaccharides was obtained from the hot-water extract of the spores of Ganoderma lucidum,and two main fractions GLP1 a and GLP1 b were isolated by DEAE Sepharose Fast Flow chromatography and Sepharose CL-6 B gel chromatography. The HPSEC analysis indicated that the two fractions were all homogeneous polysaccharides and their velative molecular mass weight were respectively 1.12 ×10^6 and 1.21 ×10^5. The analysis of composition,infrared spectrum and NMR spectroscopy indicated that GLP1 a and GLP1 b all had a backbone of β-D-（1,6）-glucan and GLP1 b were partly substituted at C-3 and C-4 in the main chain. Preliminary tests in vitro showed that the two polysaccharide fractions all has stimulating effects on murine splenocyte proliferation and the effect of GLP1 b was better than GLP1 a. Moreover,their activities were all in a dose-dependent manner. GLP1 a.