藠头凝集素的分离纯化和凝集活性分析  

Isolation, Purification and Hemagglutination Property of Lectin from Allium chinense

作　　者：肖秀情[1] 丁学知[1] 夏立秋[1] 杨琦[1] 徐沙[1] 

机构地区：[1]湖南师范大学生命科学院,微生物分子生物学省部共建国家重点实验室培育基地,湖南长沙410081

出　　处：《食品科学》2013年第19期80-83,共4页Food Science

基　　金：湖南省科技计划项目(2010FJ2002);长沙市科技计划项目(k0802102-31);湖南省高校创新平台开放基金项目(12K033)

摘　　要：藠头鳞茎经pH7.3的磷酸缓冲液抽提,80%饱和硫酸铵沉淀粗分离,Q Sepharose强阴离子交换色谱及Superdex 200 HR 10/30葡聚糖凝胶色谱分离得到藠头凝集素(Allium chinense lectin,ACL)。血细胞凝集实验显示:ACL对兔血红细胞具有高凝集活性,其最小凝集质量浓度为0.2μg/mL;在70℃热处理10min后其凝集活性保持不变,表明该凝集素具有较好的热稳定性。经100mmol/L HCl处理后ACL的凝集活性无显著变化,而在100mmol/L NaOH处理后,其凝血活性明显降低,由此表明ACL在极端碱性环境中较不稳定,但是在极端酸性环境中较稳定。甘露糖能抑制ACL对兔血红细胞的凝集作用,揭示了ACL可能与甘露糖存在特异性结合。Lectin from the bulb of Allium chinense （ACL） was isolated by extraction with phosphate buffer solution （pH 7.3）, precipitation by 80% （NH4）2SO4, and separation by Q Sepharose strong anion exchange chromatography and Superdex 200 HR 10/30 chromatography. In agglutination test, the minimum erythrocyte agglutination concentration of ACL was 0.2 μg/mL, and the hemagglutinating activity was maintained even after heating at 70 ℃ for 10 min. The agglutination activity declined after treated with 100 mmol/L NaOH but was stable in the presence of 100 mmol/L HCl, suggesting that ACL is acid-stable but not alkali-stable. The hemagglutination of the lectin on rabbit erythrocytes was inhibited by mannose among the tested sugars in this study, suggesting ACL to be a mannose binding lectin.

关 键 词：藠头 凝集素 纯化 高效液相色谱 血凝集 

分 类 号：Q946[生物学—植物学]