食品科学 ›› 2012, Vol. 33 ›› Issue (15): 226-230.

• 生物工程 • 上一篇    下一篇

韭菜过氧化物酶的分离纯化及性质

敬海明,邓 玉,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
  • 收稿日期:2011-07-29 修回日期:2012-07-04 出版日期:2012-08-15 发布日期:2012-09-07
  • 通讯作者: 唐云明 E-mail:tbright@swu.edu.cn

Isolation, Purification and Characterization of Peroxidase from Chinese Chives

  • Received:2011-07-29 Revised:2012-07-04 Online:2012-08-15 Published:2012-09-07
  • Contact: Yum-Ming Tang Yum-Ming E-mail:tbright@swu.edu.cn

摘要: 经匀浆、抽提、硫酸铵分级沉淀、CM-Sepharose 离子交换层析、Superdex-200凝胶过滤层析,获得电泳纯的韭菜过氧化物酶(POD)。该酶比活力达到14031.41U/mg,纯化倍数为102.96,回收率为10.85%。该酶分子质量约为28.4kD,最适温度40℃、最适pH值为4.6。该酶在20~40℃以及pH 4.0~8.0有较好的稳定性,在最适条件下测得其Km值为18.15mmol/L。低浓度草酸、Zn2+、Mg2+等对该酶有较强激活作用;甲醇、乙醇、异丙醇、SDS、抗坏血酸(AsA)以及Mn2+、Fe3+等对该酶有较强的抑制作用。

关键词: 韭菜, 过氧化物酶, 分离纯化, 性质

Abstract: Electrophoresis-pure peroxidase (POD) from Chinese chives was obtained after sample homogenization, extraction, ammonium sulfate precipitation, CM-Sepharose chromatography and Superdex-200 gel filtration. The purified POD had an activity of 14031.41U/mg. The purification factor was 102.96 and the recovery rate was 10.85% . The molecular weight of this enzyme was 28.4 kD, the optimum temperature and pH were 40 ℃ and 4.6, respectively. The POD enzyme was stable under 20-40℃ and pH 4-8. The Km of this enzyme was determined to be 18.15 mmol/L under optimum conditions. Its activity could be strongly activated by low concentrations of oxalic acid, Zn2+ and Mg2+,  but inhibited by methanol, ethanol, isopropanol, SDS, ascorbic acid, Mn2+ and Fe3+.

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