以石花菜为原料,对石花菜多糖进行分离纯化,并对纯化后多糖组分的理化性质及单糖组成进行分析。利用复合酶法提取石花菜多糖并采用Sevage法脱蛋白,通过DEAE-52离子交换柱层析及Sephadex G-200凝胶柱层析对粗多糖进行分离纯化,按峰收集主要的多糖组分GAP1,对其总糖、蛋白质、糖醛酸及硫酸基质量分数进行测定,并采用1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone, PMP)对多糖进行衍生化,通过高效液相色谱法测定其单糖组成。结果表明:石花菜粗多糖经柱层析后收集得到主要多糖组分GAP1,其总糖质量分数为92.56%,糖醛酸质量分数为44.53%,硫酸基质量分数为6.92%,不含蛋白质;PMP柱前衍生高效液相色谱法测得GAP1主要由鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖、木糖和L-岩藻糖6种单糖按照不同比例缩合而成,各单糖物质的量比为1.00:1.00:4.45:27.31:2.27:1.88。
To separate and purify polysaccharides from Gelidium amansii, and to analyze the physicochemical properties and monosaccharide composition of the purified polysaccharide component, GAP1. Crude polysaccharide was isolated from Gelidium amansii by multi-enzymatic extraction and deproteinized by Sevage method. The polysaccharide was fractionated and further purified into homogeneity by ion exchange chromatography on DEAE-52 column and Sephadex G-200 gel column. Its total carbohydrates, protein, uronic acid and sulfate radical contents were analyzed, and the monosaccharide composition was analyzed by high performance liquid chromatography (HPLC) after 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization. The results showed that the purified polysaccharide designated as GAP1 was obtained; the total sugar content in GAP1 was 92.56%, the uronic acid content was 44.53%, and the sulfate radical content in GAP1 was 6.92% without protein; GAP1 was composed of rhamnose, glucuronic acid, glucose, galactose, xylose and L-fucose in different ratios, the molar ratio of monosaccharide was 1.00:1.00:4.45:27.31:2.27:1.88. This study provides a theoretical foundation for the high-value utilization of Gelidium amansii polysaccharides.
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