摘要
首次报道了一步分离纯化大豆脂肪氧合酶(lipoxygenases,LOXs)的三相分离法。考察了溶剂、温度、pH值、(NH 4)2SO 4饱和度等参数对纯化倍数和活力回收率的影响。优化后的体系条件为:粗酶液与叔丁醇的体积比为1∶2.5、(NH 4)2SO 4饱和度为60%,pH值为8、温度为20℃、反应时间为60 min,其纯化倍数为8.2倍,活力回收率为78.2%。由三相法部分纯化的LOXs的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)分析显示,其分子质量约为95 kDa。结果表明三相分离法是从脱脂豆粉中部分纯化LOXs的有效方法。
Three phase partitioning(TPP)for partial purification of lipoxygenases(LOXs)in one step is reported for the first time.The influences of solvent,temperature,pH and dosage of ammonium sulfate on purification fold and activity recovery were investigated.The optimized conditions were as follows:the volume ratio of crude enzyme to tert butyl alcohol was 1∶2.5,the saturation of(NH 4)2SO 4 was 60%,the pH was 8,the temperature was 20℃,and the reaction time was 60 min.LOXs was purified by 8.2 folds and 78.2%activity was retained.SDS-PAGE analysis of partially purified enzyme revealed a molecular weight of around 95 kDa.TPP was proved to be an effective technique for the partial purification of LOXs from defatted soy flours.
作者
蔡燕
田丹
严鑫
李百裕
李宇杰
于丽娟
吴锦明
CAI Yan;TIAN Dan;YAN Xin;LI Baiyu;LI Yujie;YU Lijuan;WU Jinming(School of Chemistry and Chemical Engineering,Nantong University,Nantong 226019,China;School of Life Science,Nantong University,Nantong 226019,China;School of Pharmaceutical and Environmental Engineering,Nantong Vocational University,Nantong 226007,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2021年第9期149-153,共5页
Food and Fermentation Industries
基金
江苏省青年自然科学基金项目(BK20150401)。
关键词
三相分离
脂肪氧合酶
叔丁醇
纯化倍数
活力回收率
three phase partitioning
lipoxygenase
t-butanol
purification fold
activity recovery
