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[摘要]
目的 从膜荚黄芪根中分离纯化一种凝集素。方法 通过硫酸铵分级沉淀和ConA-Sepharose4B亲和色谱从膜荚黄芪根浸提液中分离纯化一种凝集素。结果 经过上述两步纯化得到电泳纯的凝集素,SDS-PAGE和Superdex75凝胶过滤色谱测定该凝集素的相对分子质量分别为3.15×104和3.35×104,糖蛋白染色显示其为糖蛋白,总糖的量为10.7%。结论 从膜荚黄芪根中分离纯化得到一种凝集素,该蛋白为单亚基糖蛋白,具有对兔血红细胞的凝集活性,凝集活性的比活力为391.9U/mg。
[Key word]
[Abstract]
Objective To isolate and purify a lectin from the roots of Astragalus membranaceus.Methods The protein was purified using a combination of 20%—60% ammonium sulfate fraction and ConA-Sepharose 4B affinity chromatography.Results The purified protein appeared as a single band with molecular mass of 3.15×104 on SDS-PAGE and the relative molecular mass was estimated by gel filtration on a calibrated Superdex 75 column with apparent molecular weight of 3.35×104.This lectin was a glycoprotein with a neutral carbohydrate content of 10.7%.Conclusion A lectin is isolated and purified from the roots of A.membranaceus for the first time.It is a monomer glycoprotein and its specific activity is 391.9 U/mg.
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[基金项目]
国家自然科学基金资助项目(20502032);教育部"新世纪优秀人才支持计划"(NCET-08-0534)