菜豆鲜荚中菜豆凝集素的提取与分离纯化工艺研究  

Extraction,Separation and Purification of Phaseolus vulgaris Agglutinin(PHA)from Common Bean Fresh Pods

作　　者：陈高[1] 王红波[1] 郭瑞[1] 代祥德 刘彤[1] 陈禅友[1] CHEN Gao;WANG Hongbo;GUO Rui;DAI Xiangde;LIU Tong;CHEN Chanyou(School of Life Sciences,Jianghan University;Hubei Province Engineering Research Center for Legume Plants,Wuhan 430056,Hubei,China)

机构地区：[1]江汉大学生命科学学院,湖北省豆类(蔬菜)植物工程技术研究中心,湖北武汉430056

出　　处：《江汉大学学报（自然科学版）》2016年第6期496-503,共8页Journal of Jianghan University：Natural Science Edition

基　　金：农业部948项目(2011-G1-17);湖北省科技平台项目(鄂科技通[2011]第101号);湖北省资源中心项目(2015BCE091);武汉市科技攻关项目(201250499145-11)

摘　　要：以BeanSaintespritaoeilrouge菜豆鲜荚为试验材料，优化了菜豆凝集素的提取和分离纯化工艺，研究了提取剂、料液比、pH值和提取时间对菜豆凝集素活性的影响，建立了硫酸铵分级沉淀、DEAE-52阴离子交换层析和Superdex-200凝胶过滤层析的组合方法用于分离纯化菜豆凝集素。研究结果表明：以磷酸盐缓冲溶液为提取剂，料液比1∶25，pH值6～8，浸提时间8h，菜豆植物凝集素的提取效果适宜。菜豆凝集素浸提液经硫酸铵分级沉淀，DEAE-52阴离子交换层析，Superdex-200凝胶过滤层析，冷冻干燥后可获得纯度较高的菜豆凝集素。纯化的样品经非变性聚丙烯酰氨凝胶电泳检测为单一条带，血凝法检测有血凝活性。In this paper,the processes of extraction,separation and purification of Phaseolus vulgaris agglutinin(PHA)from the common bean "Bean Saint esprit a oeil rouge" fresh pods were optimized.The effects of extracting agent,material-liquid ratio,pH and extracting time on PHA were studied,and the combined method of ammonium sulfate precipitation, DEAE- 52 anion exchange chromatography and Superdex-200 gel filtration chromatography for extracting,isolating and purifing the PHA from fresh pods of common bean has been established. The main results were as follow:1)Theoptimized technical parameters for common bean extraction process were material-liquid ratio 1∶25,pH 6-8,extracting time 8 h,PBS as extracting agent;2)The high purity PHA was obtained with ammonium sulfate precipitation,dialysis,DEAE-52 anion exchange chromatography,concentrated by ultrafiltration and Superdex-200 gel filtration chromatography. The obtained PHA shows one band with electrophoresis test and has blood clotting activity.

关 键 词：菜豆 凝集素 分离纯化 血凝活性 

分 类 号：R282.71[医药卫生—中药学] S643.1[医药卫生—中医学]