厚壳贻贝多糖MA的分离纯化、理化性质及活性研究(Ⅰ)

Isolation, purification, physicochemical properties and biological activities of polysaccharide MA from Mytilus coruscus

目的 分离纯化厚壳贻贝(Mytilus coruscus)多糖MA,并测定其理化性质和活性。方法用0.9％NaCl提取,乙醇分级沉淀,Savager法除蛋白,再经DEAE-纤维素52,Sephadex G-100柱层析,分离纯化得到多糖化合物MA。经硅胶薄层层析、聚丙烯酰胺凝胶电泳和高效液相色谱鉴定其纯度。高效液相色谱法测定其相对分子质量,过碘酸氧化,Smith降解。红外光谱和1HNMR证明其糖苷键类型,GC-MS进行糖组分分析,T、B淋巴细胞增殖反应证明其活性。结果 该糖为均一组分,相对分子质量为112 700左右。主要由葡萄糖组成,还有少量的葡萄糖胺、半乳糖胺、糖醛酸及微量的岩藻糖、甘露糖、半乳糖,且葡萄糖:葡萄糖胺:半乳糖胺:糖醛酸=74.38:10.84:8.02:6.76,其糖苷连接键为1→3,1→6和1→2糖苷键,比例约为80.390:13.434:6.176。该多糖可促进T、B淋巴细胞的增殖。结论厚壳贻贝多糖MA为不均一多糖,可促进T、B淋巴细胞的增殖。

Objective To isolate active polysaccharide MA from Mytilus coruscus and determine its physicochemical properties and bioactivities. Methods Polysaccharide MA was isolated from Mytilus coruscus by alcohol fractional precipitation, free protein was removed by Savager method. Purification of the polysaccharide by DEAE-cellulose52 and Sephadex G100 column. The homogeneity of this polysaccharide was proved by TLC, HPLC and polyacrylamide gel electrophoresis. The molecular weight was determined by HPLC and the composition of polysaccharide MA was shown by GC-MS;periodic acid oxidation,Smith degradation, IR,1HNMR spectrum indicated the kind of glycoside. Proliferation experiment of T,B lymphocytes showed the immune activities of MA. Results MA is homogenous and composed of glucose, glucamine, galactosamine, glycuronic acid, and the ratio is 74. 38:10. 84 :8. 02:6. 76. Its molecular weight is about 112700. Periodic acid oxidation,Smith degradation,IR,1 HNMR spectrum indicated the presence of many α(1→3) and a small quantity of (1→6), (1→2) linkage glycoside . MA could facilitate the proliferation of T,B lymphocytes. Conclusion MA is a kind of inhomogeneous polysaccharide, could facilitate the proliferation of T,B lymphocytes.