蜜环菌多糖的分离纯化及PMP衍生化HPLC分析

Isolation and Purification of Polysaccharides from Armillaria mellea by PMP Derivatization HPLC

目的:对蜜环菌发酵液进行多糖的提取、分离纯化,得到均一多糖,对其进行单糖组分分析。方法:分离纯化采用分步醇沉和葡聚糖凝胶(Sephadex G-200)色谱法;纯度鉴定及分子量测定采用高效液相色谱法;单糖组成采用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生HPLC法测定。结果:获得两种均一多糖(AMFP-Ⅰ、AMFP-Ⅱ),峰位分子量分别为812 939D、596217D,重均分子量(MW)分别为995 098D、872 640D,分布宽度(Mw/Mn)分别为1.268 93、1.235 20;测定单糖组成为甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖。结论:两种多糖分子量分布及单糖组成均不同,AMFP-Ⅰ主要由半乳糖醛酸、葡萄糖、半乳糖组成,AMFP-Ⅱ主要由半乳糖醛酸、半乳糖、木糖组成。

Objective： To extract,isolate and purify the polysaccharides from the Armillaria mellea fermentation broth to obtain homopolysaccharide and analyze the components.Method： The polysaccharides were isolated and purified by step alcohol precipitation and Sephadex G 200 column chromatography.The purity and molecular weight were determined by HPLC,and the monosaccharide components were determined by PMP pre-column derivatization HPLC.Result： Two kinds of homopolysaccharide（AMFP-Ⅰand AMFP-Ⅱ） were obtained with the top molecular weight of 812939D and 596217D,the weight average molecular weight（MW） of 995098D and 872640D,the distribution width（MW/Mn） of 1.268 93 and 1.235 20,respectively.The monosaccharide components were mannose,rhamnose,glucuronic acid,galacturonic acid,glucose,galactose,xylose and arabinose.Conclusion： The molecular weight,weight distribution and composition of the two monosaccharides are different.AMFP-Ⅰis composed of galacturonic acid,glucose and galactose while AMFP-Ⅱis composed of galacturonic acid,galactose and xylose.