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目的从多棘海盘车(Asterias amurensis)内脏中提取、分离纯化多糖,并对其基本理化性质和结构进行表征。方法采用沸水提取与酶解联用的方法从多棘海盘车内脏脱脂粉中提取粗多糖;采用Q Sepharose FF强阴离子交换和Sephacryl S-100凝胶渗透色谱对粗多糖进行分离纯化;采用硫酸-苯酚法、Folin-酚法、高效凝胶渗透色谱-十八角激光散射仪(HPGPC-MALLS)和高效液相色谱(HPLC)分别进行总糖含量,蛋白含量,分子量和单糖组成进行分析;采用甲基化、红外光谱(IR)、气质联用(GC/MS)和一维、二维核磁共振波谱(NMR)法对纯化多糖结构进行表征。结果从多棘海盘车内脏中提取粗多糖的收率为3.3%。经过离子交换和分子排阻色谱分离纯化,得到1种分子量均一的多糖组分F2-A,该多糖主要由葡萄糖(Glc)组成,其糖含量为97.6%,分子量为270 8kDa,是一种以α-(1(4)-Glc为主链并含有少量β-(1(3,4)分支的海星内脏来源葡聚糖。结论从多棘海盘车内脏中纯化得到1种高分子量、结构独特的葡聚糖,为将来从事其结构和生物活性关系的研究提供了有用信息。
Objective To extract, purify and purify polysaccharides from the gut of Asterias amurensis, and to characterize its basic physical and chemical properties and structure. Methods Crude polysaccharides were extracted from the visceral defatted powder of Spinonyar spinach by boiling water extraction and enzymatic hydrolysis. The crude polysaccharides were isolated and purified by Q anion exchange chromatography and Sephacryl S-100 gel permeation chromatography. The total sugar content, protein content, molecular weight and monosaccharide composition were analyzed by phenol method, Folin-phenol method, HPGPC-MALLS and HPLC respectively The structure of purified polysaccharides was characterized by methylation, IR, GC / MS and one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy (NMR). Results The yield of crude polysaccharides extracted from the spine of Spinonychids was 3.3%. After separation and purification by ion exchange and size exclusion chromatography, one kind of polysaccharide fraction F2-A with uniform molecular weight was obtained. The polysaccharide mainly consisted of glucose (Glc) with a sugar content of 97.6% and a molecular weight of 270.8 kDa, α-1 (4) -Glc was the main chain and contained a small amount of β- (1, 3,4) branched starfish-derived glucan.Conclusion One kind of high molecular weight, Unique dextran provides useful information for future studies on the relationship between its structure and biological activity.