Extraction and Partial Characterization of A Lectin from Agaricus bisporus Strain AS2796
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摘要: 双孢蘑菇2796的子实体经PBS浸提、40%～60%饱和度的硫酸铵沉淀、DEAE-Sepharose Fast Flow离子交换层析和Sephadex G-100分子筛柱层析纯化得到双孢蘑菇2796凝集素。双孢蘑菇2796凝集素经SDS-PAGE检测为单一条带,测得亚基相对分子质量为15.7 kD,通过Sephadex G-100凝胶过滤法测得其相对分子质量为64.1 kD,通过等电聚焦电泳分析发现其有4种等电点不同的亚型,pI分别为6.62、5.92、5.69、5.57。双孢蘑菇2796凝集素具有较高的热稳定性,经80℃处理10 min,仍能保持较好的凝集活性,该凝集素在pH3.0～10.0范围内都保持着较高的凝集活性,其凝血活性不依赖于二价金属离子。Abstract: A lectin was extracted and purified from the fruiting bodies of Agaricus bisporus strain 2796.The procedures included PBS extraction followed by precipitation with 40%-60%(NH4)2SO4 and filtration with DEAE-Sepharose fast flow ion-exchange chromatography and Sephadex G-100 gel filtration chromatography.The purified protein showed a band in SDS-PAGE with the subunit molecular weight of 15.7kDa.The relative molecular mass of the AS2796 lectin was 64.1kDa,as determined by the gel filtration on a Sephadex G-100 column.Four different isoelectric forms with pIs of 6.62,5.92,5.69 and 5.57 were separated by IEF-PAGE.The AS2796 lectin was stable under high temperatures.Its agglutinating activity did not decline significantly after heating at 80℃ for 10 minutes,and maintained at high level at pH 3.0-10.0.In addition,the activity was not found to be divalent metal ion-dependent.
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Key words:
- Agaricus bisporus /
- lectin /
- AS2796 /
- characterization
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