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刺槐种子中凝集素的纯化及性质研究
Purification and characterization of the Robinia pseudoacacia L.lectin
【摘要】 从刺槐(RobiniapseudoacaciaL.)的种子中分离纯化出2种凝集素RPsAⅠ和RPsAⅡ。种子经磨碎、过滤、离心、硫酸铵分级、透析、Sepharose4B离子交换和SephadexG 150分子筛层析,即可获得在PAGE和SDS PAGE上均呈现单一蛋白染色带的凝集素纯品,分子筛层析测得RPsAⅠ和RPsAⅡ的分子量分别为132000和130000,SDS PAGE测得亚基分子量分别为38000和31000。当2种凝集素的质量浓度分别达到7 80μg/mL和15 60μg/mL时能凝集兔血红细胞,凝集活性均可被L 阿拉伯糖、D 果糖、D 半乳糖、麦芽糖抑制;热稳定性实验结果表明2种凝集素对热处理均具有一定的稳定性;当pH>9和pH<5时,它们的活性都明显降低。
【Abstract】 Two lectins RPsAⅠand RPsAⅡ with strong hemagglutinating activities were isolated from the seeds of Robinia pseudoacacia L.by extraction,centrifugation,fraction with (NH4)2SO4,ion-exchange chromatogaraphy on Sepharose 4B,followed by gel filtration on Saphadex G-150.The two purified lectins showed a single protein band on PAGE.They exhibited molecular weights of 132 000 and 130 000 respectively by gel chromatogaraphy,and subunits of 38 000 and 31 000 on SDS-PAGE.They agglutinated rabbit red cell at 7.80 μg/mL and 15.60 μg/mL respectively.The reaction could be inhibited by L(+)-Arabinose,D-Fructose,D(+)-Galactose and Maltose.They’re stable to hot transaction and their activities will be detracted when pH is higher than 9.5 and lower than 5.
- 【文献出处】 江苏林业科技 ,Journal of Jiangsu Forestry Science & Technology , 编辑部邮箱 ,2004年04期
- 【分类号】Q946.1
- 【被引频次】2
- 【下载频次】85