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＋.
出处
《食品科学》
EI
CAS
CSCD
北大核心
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)
关键词
韭菜
过氧化氢酶
分离纯化
性质
Chinese chives
catalase
isolation and purification
characterization
