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珊瑚菌经热水浸提分离得到粗多糖,纯化后得到珊瑚菌多糖RBP-Ⅰ,采用凝胶渗透色谱测定其分子量,并结合Sephadex G-100层析柱鉴定其纯度。利用苯酚-硫酸法测定糖含量,碘-碘化钾法判定是否含有淀粉,考马斯亮蓝G-250法测蛋白含量,硫酸-咔唑法测糖醛酸含量。采用红外光谱扫描、气相色谱分析其结构及单糖组成。结果表明,RBP-Ⅰ的Mw为15857 Da,为均一多糖;总糖含量86.94%,糖醛酸含量5.14%,不含蛋白及淀粉;多糖RBP-Ⅰ中单糖的摩尔比为鼠李糖∶岩藻糖∶阿拉伯糖∶甘露糖∶葡萄糖∶半乳糖=1.00∶6.53∶1.99∶8.29∶70.05∶53.80;红外光谱表明其具有多糖特征峰,且为吡喃型多糖。体外抗氧化结果表明:当多糖质量浓度为10000μg/m L时,FRAP值达到0.0672 mmol/L,DPPH自由基的清除率达到22.83%,ABTS+·的清除率达到15.71%。由此可见,珊瑚菌多糖RBP-Ⅰ具有一定的抗氧化活性。
The crude polysaccharides were isolated from the coral fungus by hot water extraction. The purified polysaccharide RBP-Ⅰ was determined by gel permeation chromatography (GPC). The purity of the polysaccharides was determined by Sephadex G-100 chromatography. The content of sugar was determined by phenol-sulfuric acid method. The iodine-potassium iodide method was used to determine whether starch was contained. The protein content was determined by Coomassie Brilliant Blue G-250 method and the content of uronic acid was determined by sulfuric acid-carbazole method. The structure and the composition of monosaccharides were analyzed by infrared spectroscopy and gas chromatography. The results showed that the Mw of RBP-Ⅰ was 15857 Da, which was a homogeneous polysaccharide. The total sugar content was 86.94%, uronic acid content was 5.14%, and protein and starch were not included. The molar ratio of RBP- Fucose: arabinose: mannose: glucose: galactose = 1.00: 6.53: 1.99: 8.29: 70.05: 53.80; IR spectroscopy indicated that it has a polysaccharide characteristic peak and is a pyran type polysaccharide. In vitro antioxidant results showed that the FRAP value reached 0.0672 mmol / L, the DPPH radical scavenging rate reached 22.83%, and the ABTS + · scavenging rate reached 15.71% when the polysaccharide concentration was 10000 μg / mL. Thus, the polysaccharide RBP-Ⅰ coral polysaccharide has a certain antioxidant activity.