离子交换层析分离纯化甘薯β-淀粉酶  

Purification of β-amylase from sweet potato using ion-exchange chromatography technique

作　　者：张万利[1] 梁新红[1] 孙俊良[1] 冉军舰[1] 莫海珍[1] 

机构地区：[1]河南科技学院食品学院,河南新乡453003

出　　处：《河南科技学院学报（自然科学版）》2017年第2期23-28,共6页Journal of Henan Institute of Science and Technology(Natural Science Edition)

基　　金：河南省科技计划项目(142102110127);河南省教育厅自然科学研究资助计划项目(14A550010);河南省高校科技创新团队支持计划项目(16IRTSTHN007)

摘　　要：应用新型离子交换柱层析分离甘薯中β-淀粉酶.采用HiTrap Capto Q强阴离子柱对甘薯β-淀粉酶进行分离纯化,纯化的样品经SDS-PAGE分析纯度.结果表明:pH值为6.0的磷酸氢二钠-柠檬酸缓冲液条件下纯化效果最佳,分离色谱图显示洗脱液在280 nm的紫外吸收值最大,为496.32 m AU,蛋白峰面积最大,为225.55;洗脱液经SDS-PAGE分析为单一条带;经HiTrap Capto Q柱分离纯化后,β-淀粉酶比活力提高至96 885 U/mg,纯化倍数为15.32,酶回收率为18.30%.研究表明HiTrap Capto Q柱层析纯化能够快速高效地纯化甘薯β-淀粉酶,纯化工艺过程简单、快速、高效.研究对工业生产中运用此技术提高β-淀粉酶的分离纯化效率具有理论及实践意义.Ion-exchange chromatography technique to separate β-amylase from sweet potato was investigated.Sweet potato β-amylase were purified with a novel chromatographic technique based on the principles of anion exchange resin Capto Q with high resolution.The samples purified were identified by SDS-PAGE.The results showed that the best effect at sodium phosphate dibasic-citric acid buffer pH 6.0,and the separating chromatogram showed that the maximum ultraviolet absorption value was 496.32 m AU,the maximum peak area of protein was 225.55.The high purity β-amylase was also reflected in the single band obtained on SDS-PAGE.Under these conditions,the specific activities of was 96 885 U/mg,purification（fold） was 15.32,and yield was 18.30%.HiTrap Capto Q may be used for purification β-amylase from sweet potato,which was simple,rapid,easy to be scaled-up and suitable for large-scale production.The research has the important significance in theory and practice on high efficiently separating β-amylase from sweet potato.

关 键 词：离子交换层析 纯化 甘薯 Β-淀粉酶 

分 类 号：TS209[轻工技术与工程—食品科学] TQ925.1[轻工技术与工程—食品科学与工程]