理化因素对川泽泻凝集素血细胞凝集活性的影响  

Effect of Physical and Chemical Factors on Hemagglutination Activity of the Lectin from Alisma plantago-aquatica

作　　者：郭艳 陈松梅 张钟文 刘环燚 刘超[1] GUO Yan;CHEN Song-mei;ZHANG Zhong-wen(Applied Biotechnology Innovation Research Center,Leshan Normal University,Leshan,Sichuan 614004)

机构地区：[1]乐山师范学院应用生物技术创新研究中心,四川乐山614004

出　　处：《安徽农业科学》2023年第2期177-180,共4页Journal of Anhui Agricultural Sciences

基　　金：大学生创新创业计划国家级项目(202110649041);大学生创新创业计划省级项目(S202010649129);乐山师范学院大学生创新创业训练项目(2020XJ155);乐山师范学院重点项目(LZD011)。

摘　　要：[目的]明确温度、pH和金属离子等理化因素对川泽泻凝集素血细胞凝集生物学活性的影响。[方法]川泽泻块茎经研磨、浸取、硫酸铵沉淀、离子交换和亲和层析柱分离纯化得到凝集素(Alisma plantago-aquatica Linn.Lectin,APL),然后用系列温度、pH和金属离子等理化因素处理凝集素样品,应用倍比稀释法检测凝集素凝集人类血细胞活性。[结果]APL对家兔、家鸡、鲤鱼等动物血细胞无凝集作用,能使人ABO_(4)种类型血细胞发生不同程度凝集,对A型血细胞凝集活性明显高于其他3种血型红细胞。用20~100℃以10℃梯度处理后,川泽泻凝集素在20~40℃凝集活性很强,50℃凝集活性开始降低,在70℃仅具42%的凝集活性,80℃加热10 min完全失活;当pH为低于5.4或高于7.6时,凝集活性逐渐降低,当pH≤3.4或pH≥9.0时,川泽泻凝集素失去血细胞凝集活性;用EDTA透析后凝集素凝集活性基本消失,Ca^(2+)、Mg^(2+)、Mn^(2+)、K^(+)等离子能使凝集素凝集活性不同程度恢复。[结论]川泽泻凝集对热稳定性较弱;凝集活性最适pH为6.0~6.8,其凝集活性部分依赖于Ca^(2+)、Mg^(2+)等金属离子。[Objective]To elucidate the effects of temperature,pH and metal ions on the biological agglutination activity of lectin from Alisma planago-aquatica.[Method]Tubers of Alisma planago-aquatica were grinded,leached,precipitated with ammonium sulfate,separated and the lectin(APL)was purified by ion exchange and affinity chromatography.Then the lectin samples were treated with a series of physical and chemical factors such as temperature,pH and metal ions,and the agglutination activity to human blood cells was detected by multiple dilution method.[Result]APL had no agglutination activity on red blood cells of rabbits,domestic chickens,carp and other animals.It could agglutinate four types of human ABO red cells in different degrees.The agglutination activity of APL to type a red blood cells was significantly higher than other red blood cells.The agglutination activity of Alisma planago-aquatica lectin was very strong at 20-40℃and began to decrease at 50℃,only 42%at 70℃and complete deactivation after heating at 80℃for 10 min.When the pH was lower than 5.4 or higher than 7.6,the agglutination activity decreased gradually.When the pH was lower than 3.4 or higher than 9.0,the agglutination activity of Alisma planago-aquatica lectin lost.After dialysis with EDTA,the agglutination activity of lectin basically disappeared.The agglutination activity of lectin could be restored to some extent by different ions such as Ca^(2+),Mg^(2+),Mn^(2+),K^(+),and so on.[Conclusion]The agglutination activity of Alisma planago-aquatica lectin is weak to thermal stability,and the optimal pH is 6.0-6.8.The agglutination activity is partly dependent on Ca^(2+),Mg^(2+)and other metal ions.

关 键 词：川泽泻凝集素 血细胞 理化因素 凝集活性 

分 类 号：Q513[生物学—生物化学]