红栓菌固培物多糖的分离纯化与分析

Isolation, Purification and Analysis of Polysaccharides from PCM

红栓菌(Pycnvporus cinnabarins)菌丝固体培养物经热水提取,H_2O_2氧化法脱色素,Sevage法除蛋白,再通过DEAE-Sephadex A-25分离及Sephadex G-200进一步纯化,得到三种多糖,命名为PCM_1、PCM_2、PCM_3。三者经聚丙烯酰胺电泳、超速离心及Sephadex G-200层析鉴定,为均一体。紫外光谱分析未见有核酸与蛋白质的特征吸收峰,红外光谱及核磁共振氢谱分析提示:三者分子中的糖苷键为α型。组成分析表明:PCM_1由D-葡萄糖、D-半乳糖及D-甘露糖组成;PCM_2由D-葡萄糖、D-半乳糖组成;PCM_3为甘露聚糖。分子量依次为17500,7800,5600。

PCM is the code name of the substance which consists of Pycnoporus cinnabarius mycelium and its solid culture medium. A polysaccharide was obtained by extraction with hot water and precipitated by alcohol,and futher isolation and purification steps included depigment by H_2O_2 oxidation method, deprotein by Sevage method,dialysis, chromatography on DEAE-Sephadex A-25 and Sephadex G-200. Three fractions were obtained and denoted as PCM_1, PCM_2 and PCM_3. They were identified to be single substance by gel chromatography, PAGE and ultracentrifugation. The molecular weight was 17500,7800 and 5600 respectively. Three polysaccharides showed the positive optical rotation and did. not contain nitrogen. The IR of them showed absorption bands to be characteristic of polysaccharide. ~1H-NMR estimation revealed that there was a-glycosidic linkage in molecules. According to the results of PC and GC: PCM_1 was composed of D-glucose, D-galactose and mannose, PCM_2 was composed of D-glucose and D-galactose, and PCM_3 was mannan.