[关键词]
[摘要]
本文以秀珍菇菌丝体为原料提取水溶性粗多糖,经脱蛋白、脱色、DEAE-Sepharose F.F和Superdex-200柱层析纯化,得到一种秀珍菇菌丝体多糖(PMP-2a)。高效液相色谱鉴定其纯度;比色法测定其总糖、糖醛酸、蛋白质含量;紫外光谱、红外光谱、β-消去等方法研究其理化性质和部分结构;通过清除超氧阴离子自由基、羟基自由基来研究PMP-2a的体外抗氧化活性。结果表明,PMP-2a为均一组分,其中性糖部分由鼠李糖(Rha)、葡萄糖(Glc)、半乳糖(Gal)、甘露糖(Man)、阿拉伯糖(Ara)组成(摩尔比为1:13.18:2.19:7.23:3.15);其总糖、糖醛酸、蛋白质含量分别为82.6%、19.8%、10.6%,不含有核酸;红外光谱呈现典型的多糖特征峰,含有α-D-吡喃糖苷键;相对分子质量为9.28×104 Da,β-消去反应表明糖链与蛋白之间存在-O-的连接键;PMP-2a对超氧阴离子自由基、羟基自由基的清除效果显著,且与多糖浓度呈明显的量效关系。
[Key word]
[Abstract]
In this study, a water-soluble, crude polysaccharide was extracted from Pleurotus geesteranus and the mycelium polysaccharide was obtained by deproteinization, discoloration, and purification using diethylaminoethanol (DEAE)-Sepharose Fast-Flow ion-exchange chromatography and Superdex-200 column chromatography. The purity of the polysaccharide was determined using high-performance liquid chromatography (HPLC); Colorimetric measurement of the total sugar, uronic acid and protein content; and physicochemical properties and partial structural characteristics, studied by ultraviolet (UV), infrared (IR) and β-elimination reaction methods. The in vitro antioxidant activity of PMP-2a was determined by superoxide radical and hydroxyl radical scavenging assays. The results showed that PMP-2a was a homogeneous polysaccharide, consisting of Rha, Glc, Gal, Man, and Ara in a molar ratio of 1:13.18:2.19:7.23:3.15. The total sugar, uronic acid, and protein content of PMP-2a were 82.6%, 19.8%, and 10.6%, respectively; no nucleic acid was found. Typical absorption bands of polysaccharides and glycosidic bonds in α-D-pyranoside were found in the IR spectrum. The molecular weight was 9.28 × 104 Da, and the β-elimination reaction indicated that PMP-2a was linked to peptide chains through an -O- bond. PMP-2a showed significant superoxide radical- and hydroxyl radical-scavenging activities in a dose-dependent manner.
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[基金项目]
安徽省高等学校省级自然科学研究重点项目(KJ2014A052);国家自然科学基金项目(31100070);2013年国家级大学生创新创业训练计划项目(201310879021)