食品科学 ›› 2002, Vol. 23 ›› Issue (9): 49-53.

• 基础研究 • 上一篇    下一篇

蜜环菌多糖分离纯化及性质的研究

 谭周进, 谢达平, 王征, 肖启明, 李立恒   

  1. 湖南农业大学食品科技学院
  • 出版日期:2002-09-15 发布日期:2011-12-31

Studies on Isolation ,Purification and some Properties of Polysaccharide from Armillaria Mellea

 TAN  Zhou-Jin, XIE  Da-Ping, WANG  Zheng, XIAO  Qi-Ming, LI  Li-Heng   

  • Online:2002-09-15 Published:2011-12-31

摘要: 进行了蜜环菌胞外多糖分离纯化及性质的研究,得到如下结果:发酵液浓缩,三倍体积乙醇沉淀、75%乙醇洗涤、Sevage法除蛋白、20%H2O2脱色,得到的粗多糖上DEAE-纤维素(OH-)柱层析,用蒸馏水、0.05~0.5mol/L的NaCl梯度洗脱,可以得到中性多糖和几种酸性多糖。中性多糖上SephadexG-150柱层析,用蒸馏水洗脱,得到一种多糖A,多糖A为纯多糖。在400~4000cm-1范围内摄得多糖A的红外光谱表明其含有β-糖苷键。多糖A的完全酸水解液用硅胶GF254薄层层析,氯仿-甲醇(60:40)与丙酮-水(96:4)展层,以及纸层析,丙酮-水(96:4)展层,苯胺-二苯胺显色,证明其组成单糖为葡萄糖。考马斯亮兰G-250反应阴性、茚三酮反应阴性、双缩脲反应阴性,聚丙烯酰胺凝胶电泳后、考马斯亮兰R-250染色无蛋白质带,都证明其不含蛋白质。

关键词: 蜜环菌, 深层发酵, 胞外多糖, 分离纯化, 多糖性质

Abstract: The study on isolation and property of exopolysaccharides from Armillaria mellea was conducted.The following results were obtained : The fermentation broth was concentrated. Then alcohol was put thrice into it .The precipitate attained was eluted again with 75% alcohol .The protein was eliminated with Sevage method.Furthermore,the DEAE-cellulose column chromatography was used on the exopolysaccharide purification and the column was eluted with the distilled water and NaCl solution from 0.05~0.5mol/L.Neutral and several kinds of acid polysaccharides were obtained . The purification of the neutral polysaccharide was made with SephadexG-150 chromatography.The pure polysaccharide A was collected. The IR spectrum of the polysaccharide A showed a character of β-glycosidic bond in 400~4000cm-1.After the polysaccharide A was hydrolyzed, the hydrolysate was analyzed with the silica gel pellicular chromatography and filter paper chromatography . While both the chloroform:methanol(60:40) and acetone:H2O(96:4) were used as the disperse agent and the dische reaction was used as dyestuff, the results proved that polysaccharide was composed of glucose. The polysaccharide was colored by coomassie brilliant blue G-250, and coomassie brilliant blue R-250 after polysaccharide electrophoresis. It proved that there was not any protein remained in the polysaccharide A.

Key words: Armillaria mellea , Submerged fermentation , Exopolysaccharide , Isolation and purification , Polysaccharide property