利用中国被毛孢虫草菌液体深层发酵培养得到的菌丝体,通过正交试验优化得到了虫草多糖的提取工艺条件为:提取温度100℃,提取时间120 min,料液比1∶15,提取次数4次。粗多糖经Sevag法脱蛋白、透析袋透析、DEAE-cellulose离子交换和Sephadex G-100柱层析纯化后,得到3种多糖(命名为HSP-1、HSP-2和HSP-3)。经紫外扫描、Sephacryl S-300 HR柱层析检测确定HSP-1和HSP-2为均一组分,HSP-3为非均一组分。苯酚-硫酸法测得HSP-1和HSP-2的含糖量分别为89.42%和85.63%。Sephacryl S-300 HR柱层析测得HSP-1和HSP-2的相对分子质量分别为1.7×104Da和9.8×103Da。GC-MS测得HSP-1的单糖组成及比例为:D-葡萄糖∶D-甘露糖∶D-半乳糖=4.522∶1∶1.378,HSP-2的单糖组成及比例为:D-葡萄糖∶D-甘露糖∶D-半乳糖∶D-阿拉伯糖=2.687∶4.591∶1∶0.212。红外光谱测定结果显示HSP-1和HSP-2都含有α型糖苷键。

The mycelia were acquired by liquid submerged fermentation with Hirsutella sinensis. The extraction conditions were optimized through orthogonal test as follows: the extraction temperature was 100 ℃,the extraction time was120 min,and the ratio of solid to liquid was 1∶ 15,for four times extraction. The crude polysaccharide was stepwise purified by Sevag deproteinization,dialysis,DEAE cellulose Ion exchange and Sephadex G-100 column chromatography,and three kinds of polysaccharides( named HSP-1,HSP-2,and HSP-3) were obtained. After ultraviolet scanning and Sephacryl S-300 HR column determination,HSP-1 and HSP-2 were regarded as homogeneous components while HSP-3 was inhomogeneous. The results from Phenol-sulfuric acid determination showed that the contents of HSP-1 and HSP-2 were 89. 42% and 85. 63%,respectively. The average molecular weights of HSP-1 and HSP-2,which were determined by Sephacryl S-300 HR column chromatography,were estimated to be 1. 7 × 104Da and 9. 8 × 103Da,respectively. The results of GC-MS showed that monosaccharide compositions of HSP-1 contained D-glucose,D-mannose and D-galactose with molar ratio of 4. 522 ∶ 1 ∶ 1. 378,while monosaccharide compositions of HSP-2 were D-glucose,D-mannose,D-galactose and D-arabinose with molar ratio of 2. 687∶ 4. 591∶ 1∶ 0. 2121. The FT-IR detection showed that the α-configuration glycosylic bond was contained in both HSP-1and HSP-2.