藠头凝集素的分离纯化和凝集活性分析 - 食品科学.pdf

80 2013, Vol.34, No.19 食品科学 ※基础研究 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 (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) 摘  要： pH7.3 80 Q Sepharose 藠头鳞茎经 的磷酸缓冲液抽提， %饱和硫酸铵沉淀粗分离， 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 0.2μg/mL 70 10min 对兔血红细胞具有高凝集活性，其最小凝集质量浓度为 ；在 ℃热处理 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。 关键词：藠头；凝集素；纯化；高效液相色谱；血凝集 Isolation, Purification and Hemagglutination Property of Lectin from Allium chinense XIAO Xiu-qing ，DING Xue-zhi*，XIA Li-qiu ，YANG Qi ，XU Sha (State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha 410081, China) Abstract ：Lectin from the bulb of Allium chinense (ACL) was isolated by extraction with phosphate buffer solution (pH 7.3), precipitation by 80% (NH ) SO , and separation by Q Sepharose strong anion exchange chromatography and Superdex 4 2 4 200 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 hemaggluti