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目的通过醇沉、色谱等分离提取技术对发酵得到的亮菌胞内多糖进行分离纯化,并对纯化的多糖进行结构及部分性质研究。方法采用水提、醇沉、除蛋白等方法得到亮菌胞内粗多糖,然后通过离子交换色谱、分子筛色谱对粗多糖进行进一步的分离纯化,得到单一多糖。同时采用红外光谱法、紫外光谱法、核磁共振法对该单一多糖进行结构研究,确定其单糖组成及糖链的连接方式。结果通过去蛋白、醇沉、离子交换色谱和凝胶滤过色谱的方法得到亮菌胞内多糖IPS-B2,该多糖为直链α-D-(1→6)-吡喃型葡聚糖,不含蛋白质、多肽和氨基酸。结论通过纯化得到了直链α-(1→6-葡聚糖(IPS-B2),该多糖在体内具有较好的抑瘤效果。
OBJECTIVE To separate and purify the breve polysaccharides obtained from the fermentation by alcohol-settling, chromatographic separation, and to study the structure and some properties of the purified polysaccharides. Methods The crude polysaccharide was obtained by water extraction, alcohol precipitation and protein removal. Then the polysaccharide was further purified by ion exchange chromatography and molecular sieve chromatography to obtain a single polysaccharide. At the same time, infrared spectroscopy, ultraviolet spectroscopy and nuclear magnetic resonance were used to study the structure of the single polysaccharide, and its monosaccharide composition and sugar chain attachment were determined. The result was deproteinized, alcohol precipitation, ion exchange chromatography and gel filtration chromatographic methods to obtain the bright bacteria intracellular polysaccharide IPS-B2, which is a linear α-D-(1→6)-glucopyranose. , Contains no proteins, peptides and amino acids. Conclusion The purified α-(1->6-glucan (IPS-B2)) was obtained through purification. The polysaccharide has good anti-tumor effect in vivo.