海带硫酸多糖的提取、纯化及其理化分析

Extraction,purification and physicochemical analysis of Laminaria polysaccharide sulfate

目的研究海带硫酸多糖 (LPS)的提取制备及其组分的纯化和理化性质。方法采用酶解法从海带提取制备LPS,海带浆加入纤维素酶、半纤维素酶、果胶酶和蛋白酶 ,50℃水解 4h。取滤液加入氯化钙 ,离心去除海藻酸钙。取上清液加入十六烷基三甲基溴化铵 (CTAB)与LPS结合沉淀 ,离心收集CTAB LPS沉淀物。加入氯化钙溶液进行盐解 ,将LPS游离释放出来 ,加入乙醇使LPS析出。离子交换色谱和凝胶过滤色谱法分离纯化其多糖组分。结果酶解提取法的收率为 5‰ ,色谱法分离得到 4个主要多糖组分 ,测定其分子量分别为 2 1 0、1 2 0、40 0和 1 4 0kD。酶解法的提取收率高于水煮法。

PurposeTo study extraction,purification and physicochemical analysis of Laminaria polysaccharide sulfate(LPS).MethodsThe LPS was extracted from Laminaria japonica. Laminaria japonica was crushed to slurry, to which cellulase, semicellulase, pectase and protease was added and the enzymolysis was carried out at 50℃ for 4 h. Then the resulted mixture was filtered and CaCl 2 was added to the filtrate to remove alginic acid. CTAB was added to supernatant and CTAB LPS precipitate was obtained. At higher concentration of CaCl 2, LPS was released from CATB LPS and then precipitated with alcohol. LPS was obtained with centrifugation and freeze drying. Ion exchange chromatography and gel filtration chromatography were per formed to isolate and purify the crude LPS.ResultsThe productivity of LPS was 5‰ by enzymolysis. Four fractions of LPS were obtained by chromatography and their molecular weights were 210,120,400 and 140 kD respectively.ConclusionThe productivity of LPS by enzymolysis was higher than that by water boiling method.