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Purification and enzymatic properties of a peroxidase from leaves of Phytolacca dioica L. (Ombú tree)

Vincenzo Guida|Giovanna Criscuolo|Rachele Tamburino|Livia Malorni|Augusto Parente|Antimo Di Maro
생화학분자생물학회 (Korean Society Of Biochemistry And Molecular Biology)|2011.01|BMB Reports|제44권 제1호|pp. 64-69

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키워드

Protein purificationKineticsPeroxidaseEnzymatic activityPhytolacca dioica
주제분류화학

초록

A peroxidase (PD-cP; 0.47 mg/100 g leaves) was purified from autumn leaves of Phytolacca dioica L. and characterized. PD-cP was obtained by acid precipitation followed by gel-filtration and cation exchange chromatography. Amino acid composition and N-terminal sequence of PD-cP up to residue 15 were similar to that of Spinacia oleracea (N-terminal pairwise comparison showing four amino acid differences). PD-cP showed a molecular mass of approx. 36 kDa by SDS-PAGE,pH and temperature optima at 3.0 and 50.0oC, respectively and seasonal variation. The Michaelis-Menten constant (KM) for H2O2 was 5.27 mM, and the velocity maximum (Vmax) 1.31nmol min-1, while the enzyme turnover was 0.148 s-1. Finally,the presence of Ca2+ and Mg2+ enhanced the PD-cP activity,with Mg2+ 1.4-fold more effective than Ca2+ [BMB reports 2011; 44(1): 64-69]

Abstract

A peroxidase (PD-cP; 0.47 mg/100 g leaves) was purified from autumn leaves of Phytolacca dioica L. and characterized. PD-cP was obtained by acid precipitation followed by gel-filtration and cation exchange chromatography. Amino acid composition and N-terminal sequence of PD-cP up to residue 15 were similar to that of Spinacia oleracea (N-terminal pairwise comparison showing four amino acid differences). PD-cP showed a molecular mass of approx. 36 kDa by SDS-PAGE,pH and temperature optima at 3.0 and 50.0oC, respectively and seasonal variation. The Michaelis-Menten constant (KM) for H2O2 was 5.27 mM, and the velocity maximum (Vmax) 1.31nmol min-1, while the enzyme turnover was 0.148 s-1. Finally,the presence of Ca2+ and Mg2+ enhanced the PD-cP activity,with Mg2+ 1.4-fold more effective than Ca2+ [BMB reports 2011; 44(1): 64-69]

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