食品科学 ›› 2011, Vol. 32 ›› Issue (9): 217-221.doi: 10.7506/spkx1002-6630-201109049

• 生物工程 • 上一篇    下一篇

韭菜过氧化氢酶的分离纯化及其部分酶学特性

邓 玉,敬海明,成丽丽,赵 芯,刘玉杰,唐云明*   

  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
  • 出版日期:2011-05-15 发布日期:2011-04-11
  • 基金资助:
    重庆市科委科技攻关项目(CSCT2004AC1012)

Isolation, Purification and Partial Characterization of Catalase from Chinese Chives

DENG Yu,JING Hai-ming,CHENG Li-li,ZHAO Xin,LIU Yu-jie,TANG Yun-ming*   

  1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweetpotato Engineering Research Center, School of Life Science, Southwest University, Chongqing 400715, China
  • Online:2011-05-15 Published:2011-04-11

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。

关键词: 韭菜, 过氧化氢酶, 分离纯化, 性质

Abstract: 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, fractional 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+.

Key words: Chinese chives, catalase, isolation and purification, characterization

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