SCI和EI收录∣中国化工学会会刊

›› 2010, Vol. 18 ›› Issue (5): 824-829.

• • 上一篇    下一篇

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

陈敏1,2, 姚善泾1, 张虹1, 梁新乐2   

  1. 1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;
    2. Department of Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310012, China
  • 收稿日期:2009-12-29 修回日期:2010-05-01 出版日期:2010-10-28 发布日期:2010-10-28
  • 通讯作者: YAO Shanjing,E-mail:yaosj@zju.edu.cn E-mail:yaosj@zju.edu.cn
  • 基金资助:
    Supported by the Special Funds for Major State Basic Research Program of China (2007CB707805);the Natural Science Foundation of Zhejiang Province (Y505334)

Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii

CHEN Min1,2, YAO Shanjing1, ZHANG Hong1, LIANG Xinle2   

  1. 1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;
    2. Department of Biotechnology Engineering, Zhejiang Gongshang University, Hangzhou 310012, China
  • Received:2009-12-29 Revised:2010-05-01 Online:2010-10-28 Published:2010-10-28
  • Supported by:
    Supported by the Special Funds for Major State Basic Research Program of China (2007CB707805);the Natural Science Foundation of Zhejiang Province (Y505334)

摘要: A versatile peroxidase (VP-Peco60-7) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP-Peco60-7 in this study is of a novel type. With 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50℃ and pH 3.0. The catalysis of ABTS by VP-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50℃ and pH 3.0, the maximum velocity (Vmax) was 188.68 U·mg-1 and the michaelis constant (Km) was 203.09 μmol·L-1.

关键词: Pleurotus eryngii, versatile peroxidase, purification, enzymatic properties

Abstract: A versatile peroxidase (VP-Peco60-7) was generated and purified from the liquid culture of Pleurotus eryngii. The purification procedure included ammonium sulfate precipitation, ion exchange chromatography, and gel chromatography. The molecular weight and isoelectric point (pI) of VP-Peco60-7 were determined to be approxi-mately 40 kDa and 4.1, respectively. By N-terminal sequence determination and peptide mapping analysis, VP-Peco60-7 was found to be similar to the versatile peroxidase isoenzyme VPL1, which was previously isolated from liquid cultures of the same species. However, the molecular weight and pI of VP-Peco60-7 were different from those of versatile peroxidases of liquid cultures, implying that the VP-Peco60-7 in this study is of a novel type. With 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as a substrate, the maximal enzyme activity was obtained at 50℃ and pH 3.0. The catalysis of ABTS by VP-Peco60-7 was expressed by the Michaelis-Menten equa-tion. At 50℃ and pH 3.0, the maximum velocity (Vmax) was 188.68 U·mg-1 and the michaelis constant (Km) was 203.09 μmol·L-1.

Key words: Pleurotus eryngii, versatile peroxidase, purification, enzymatic properties