霍山铁皮石斛糖蛋白的制备及抗氧化活性  

Purification and antioxidant activity of glycoprotein fromHuoshan Dendrobium officinale

作　　者：何晓梅 徐海军 邓辉 张珍林 孙传伯 谷仿丽 余茂耘 HE Xiaomei;XU Haijun;DENG Hui;ZHANG Zhenlin;SUN Chuanbo;GU Fangli;YU Maoyun(Anhui Engineering Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources,College of Biology and Pharmaceutical Engineering,West Anhui University,Liu′an 237012,China)

机构地区：[1]皖西学院生物与制药工程学院安徽省中药资源保护与持续利用工程实验室,安徽六安237012

出　　处：《生物加工过程》2020年第3期312-317,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家中药材产业技术体系(CARS 21);安徽省自然科学基金(1808085MH307);安徽省教育厅重点项目(KJ2016A887、KJ2017A407、KJ2018A0420、KJ2017A403、KJ2019A0627、KJ2019A0625)。

摘　　要：以霍山铁皮石斛枫斗为试验材料,经脱脂脱色、0.3 mol/L盐溶液浸提、Sevage法脱蛋白、硫酸铵分级沉淀、透析、聚乙二醇10000浓缩、冷冻干燥得到粗品;经SDS-PAGE电泳、DEAE 52离子柱和Sephadex G-100凝胶柱分离纯化得到糖蛋白并测定其抗氧化活性。SDS-PAGE电泳显示硫酸铵饱和度越大,盐析作用越强,糖蛋白条带越明显,糖蛋白分子量在6.62×10^4附近;DEAE 52离子柱和Sephadex G-100凝胶柱逐步分离纯化得到2个组分的糖蛋白GP1和GP2;分光光度法测定GP1和GP2中糖蛋白含量分别约为95%和87%;抗氧化结果表明,霍山铁皮石斛糖蛋白具有较强的还原能力且可有效地清除自由基·DPPH、·OH和·O2^-。We used Huoshan Dendrobium officinale as raw material to extract crude glycoprotein with 0.3 mol/L saline solution,followed by deproteinization,ammonium sulfate precipitation,concentration,dialyzing and freeze-drying.Polyethylene glycol(PEG)was used as the control.The purified glycoprotein was then obtained with SDS-PAGE electrophoresis,DEAE-52 cellulose and Sephadex-100,and its antioxidant activity was determined.Higher degree of ammonium sulfate saturation resulted in better salting-out effect.The relative molecular mass is round 6.62×10^4.Two glycoproteins were obtained through DEAE-52 cellulose and Sephadex-100.Content of sugar and protein in glycoprotein was determined as 5.65% and 0.017% respectively through spectrophotometry.The glycoprotein was reductive and effective for clearing up DPPH free radicals,hydroxyl radical,and superoxide anion.

关 键 词：霍山铁皮石斛 糖蛋白 SDS PAGE电泳 制备 抗氧化 

分 类 号：TS24[轻工技术与工程—制糖工程]