食品科学

• 基础研究 • 上一篇    下一篇

藠头凝集素的分离纯化和凝集活性分析

肖秀情,丁学知*,夏立秋,杨 琦,徐 沙   

  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
  • 发布日期:2013-09-27
  • 通讯作者: 丁学知
  • 基金资助:

    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)

Isolation, Purification and Hemagglutination Property of Lectin from Allium chinense

XIAO Xiu-qing,DING Xue-zhi*,XIA Li-qiu,YANG Qi,XU Sha     

  1. State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University,
    Changsha 410081, China
  • Published:2013-09-27
  • Contact: DING Xue-zhi

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/LNaOH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。

关键词: 藠头, 凝集素, 纯化, 高效液相色谱, 血凝集

Abstract:

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.

Key words: Allium chinense, lectin, purification, high performance liquid chromatography (HPLC), hemagglutination

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