桦褐孔菌多糖的纯化及其单糖组成分析

Purifi cation and monosaccharide composition of polysaccharides from Inonotus

目的对桦褐孔菌多糖进行纯化,并对其中的单糖组成进行研究。方法桦褐孔菌粗多糖经过脱蛋白、脱色得到纯化多糖。柱前衍生化后利用气相色谱分析得到多糖的单糖组成。结果酶法+TCA法蛋白脱除率较高,为82.32%;多糖保留较多,为76.13%。聚酰胺层析柱法多糖脱色效果最好,脱色率高达93.39%,并且多糖保留率也高达81.68%。多糖脱蛋白后的纯度从之前的21.5%提升到80.4%,脱色后的纯度从80.4%提升到85.2%。桦褐孔菌的多糖主要由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖组成,各单糖质量百分数分别占3.84%、3.69%、3.32%、18.52%、27.30%、28.17%。结论桦褐孔菌多糖最佳脱蛋白方法为酶法+TCA法,最适宜的脱色方法是聚酰胺层析柱法。气相色谱分析方法简便、灵敏,不但多糖纯度高,而且可以准确测出桦褐孔菌多糖中单糖组成。

Objective To purify and analyze the monosaccharide composition of polysaccharides from Inonotus. Methods Crude polysaccharides from Inonotus were purifi ed into fractions by removing protein and bleaching. After pre-column derivatization, gas chromatography was used to analyze the monosaccharide composition. Results Enzymatic and TCA method was used to remove the protein, with the removal rate of 82.32% and polysaccharide retention of 76.13%. Polyamide column chromatography method was used best of bleaching, with decoloring rate of 93.39%, and polysaccharide retention rate of 81.68%. The monosaccharide composition of Inonotus polysaccharides was comprised of rhamnose, arabinose, xylose, mannose, glucose and galactose at mass percent of 3.84%, 3.69%, 3.32%, 18.52%, 27.30% and 28.17%. Conclusion The best way to remove the protein in polysaccharides is enzymatic and TCA method, and the most optimum bleaching method is polyamide column chromatography. The method of gas chromatography is simple and sensitive, and can be used for the analysis of monosaccharide hydrolyzed from Inonotus polysaccharide.