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

Abstract

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 (V_max) was 188.68 U·mg^-1 and the michaelis constant (K_m) was 203.09 μmol·L^-1.

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

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 (V_max) was 188.68 U·mg^-1 and the michaelis constant (K_m) was 203.09 μmol·L^-1.

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

CHEN Min, YAO Shanjing, ZHANG Hong, LIANG Xinle. Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii[J]. , 2010, 18(5): 824-829.

陈敏, 姚善泾, 张虹, 梁新乐. Purification and Characterization of a Versatile Peroxidase from Edible Mushroom Pleurotus eryngii[J]. , 2010, 18(5): 824-829.

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