生物学杂志

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霍山石斛糖蛋白的提取方法优选及抗氧化活性分析

  

  1. 1. 皖西学院 生物与制药工程学院, 六安 237012; 2. 中科院云计算中心公共安全研究院, 东莞 523808; 3. 安徽省中药资源保护与持续利用工程实验室, 六安 237012;4. 安徽省石斛产业化开发协同创新中心, 六安 237012
  • 出版日期:2019-04-18 发布日期:2019-04-18
  • 通讯作者: 陈乃富,教授,研究方向为天然产物和中药资源开发,E-mail:cnf505@126.com
  • 作者简介:邓辉,博士,研究方向为中药标准化、蛋白质和酶工程,E-mail:dhup@qq.com
  • 基金资助:
    国家自然科学基金(81573536);安徽省科技厅自然科学基金(1808085MH307);安徽省杰出青年科学基金项目(1808085J17);2017安徽高校优秀青年人才支持计划项目(gxyq2017057);皖西学院自然科学基金(WXZR201616,WXZR201816)

Optimization of extract method and analysis of antioxidant activity of glycoprotein from Dendrobium huoshanense

  1. 1. College of Biology and Pharmaceutical Engineering, West Anhui University, Lu′an 237012; 2. Institute ofPublic Safety, Cloud Computing Center, Chinese Academy of Sciences, Dongguan 523808; 3. Anhui Engineering  Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources, Lu′an 237012;4. Anhui Province Synergy Innovation Center of Industrialization Development of Dendrobium, Lu′an 237012, China
  • Online:2019-04-18 Published:2019-04-18

摘要: 以道地药材——霍山石斛鲜茎为试验材料,分别按提取蛋白的盐溶液浸法和提取多糖的水提醇沉法,在4℃条件下提取糖蛋白,考察比对其提取效果,以及不同糖蛋白组分的抗氧化活性。由两种方法获得的提取液分别经硫酸铵分级沉淀和乙醇分级沉淀,获得6组糖蛋白提取液,经SDS-PAGE电泳,蛋白染色和糖染色确定提取效果。结果表明:盐溶液浸提法获得的糖蛋白谱带在浓度、数量和分子量范围上都显著高于水提醇沉法。体外抗氧化试验表明,谱带多态性较高的糖蛋白提取液具有较高的清除超氧阴离子自由基和羟基自由基的能力。实验进一步通过单因素和正交试验确定了盐溶液浸提霍山石斛糖蛋白的最佳条件:NaCl 溶液浓度0.3 mol/L、液料比16∶ 1、提取时间12 h。在此条件下,粗糖蛋白得率(12.56±0.35)%。

关键词: 霍山石斛, 糖蛋白, 提取方法, 抗氧化活性

Abstract: Under 4℃, the extracted liquid was obtained from Dendrobium huoshanense′s fresh stems with graded precipitated by ammonium sulfate and ethanol, respectively, and six kinds of glycoprotein extract components were got and determined by SDS-PAGE, protein staining and sugar staining. The results showed that the glycoprotein bands obtained by the salt solution were significantly higher in concentration, quantity and molecular weight range. In vitro antioxidant experiments showed that the glycoprotein extract with high polymorphism bands had higher capacity to scavenging superoxide anion free radical and hydroxyl radical. The optimal extraction conditions in saline solution of glycoprotein from Dendrobium huoshanense were 0.3 mol/L NaCl solution, at the liquid-solid ratio of 16∶ 1, extraction for 12 h, and the yield rate of crude glycoprotein was (12.56±0.35)%.

Key words: Dendrobium huoshanense, glycoprotein, extraction method, antioxidant activity

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