韭菜过氧化氢酶的分离纯化及其部分酶学特性  

Isolation, Purification and Partial Characterization of Catalase from Chinese Chives

作　　者：邓玉[1] 敬海明[1] 成丽丽[1] 赵芯[1] 刘玉杰[1] 唐云明[1] 

机构地区：[1]西南大学生命科学学院,重庆市甘薯工程研究中心,三峡库区生态环境教育部重点实验室,重庆400715

出　　处：《食品科学》2011年第9期217-221,共5页Food Science

基　　金：重庆市科委科技攻关项目(CSCT2004AC1012)

摘　　要：为获得韭菜过氧化氢酶纯品并对其性质进行研究,将新鲜韭菜通过匀浆、抽提、硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Superdex-200凝胶过滤层析等步骤,获得了电泳纯的韭菜过氧化氢酶(CAT),纯化倍数为70.36,回收率为18.33%,酶比活力达到22064.57U/mg。其全酶分子质量和亚基分子质量分别为241.76、62.43kD。该酶反应的最适温度为37℃,最适pH值为7.2。该酶在25～40℃以及pH5～9有较好的稳定性,同时在最适条件下测得其Km值为46.93mmol/L。甲醇、乙醇、异丙醇、SDS以及Cu2+、Ag+、Fe2+等金属离子对该酶有较强的抑制作用。The present study was conducted to obtain high-purity catalase（CAT） from Chinese chives and explore some of its enzymological properties.Electrophoresis-purity CAT was obtained sequentially after homogenization,extraction,frac-tional ammonium sulfate precipitation,DEAE-Sepharose chromatographic separation and Superdex-200 gel filtration.In the process,a purification factor of 70.36,a recovery of 18.33% and a specific enzyme activity of 22064.57 U/mg were obtained.The enzyme exhibited a molecular weight of 241.76 kD and contained a 62.43 kD subunite.The optimum temperature and pH for this enzyme were 37 ℃ and 7.2,respectively.The CAT enzyme was stable under pH 5-9 and 25-40 ℃ conditions.Its Km was determined to be 46.93 mmol/L under optimum conditions.The enzyme activity could be strongly inhibited by methanol,ethanol,isopropanol,SDS,Cu2＋,Ag＋,and Fe2＋.

关 键 词：韭菜 过氧化氢酶 分离纯化 性质 

分 类 号：Q946.5[生物学—植物学]