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

  • Guida, Vincenzo (Dipartimento di Scienze della Vita, Seconda Universita di Napoli) ;
  • Criscuolo, Giovanna (Dipartimento di Scienze della Vita, Seconda Universita di Napoli) ;
  • Tamburino, Rachele (Dipartimento di Scienze della Vita, Seconda Universita di Napoli) ;
  • Malorni, Livia (Centro di Spettrometria di Massa Proteomica e Biomolecolare, Istituto di Scienze dell'Alimentazione del CNR) ;
  • Parente, Augusto (Dipartimento di Scienze della Vita, Seconda Universita di Napoli) ;
  • Maro, Antimo Di (Dipartimento di Scienze della Vita, Seconda Universita di Napoli)
  • Received : 2010.07.21
  • Accepted : 2010.12.08
  • Published : 2011.01.31
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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 $50.0^{\circ}C$, respectively and seasonal variation. The Michaelis-Menten constant ($K_M$) for $H_2O_2$ was 5.27 mM, and the velocity maximum ($V_{max}$) $1.31\;nmol\;min^{-1}$, while the enzyme turnover was $0.148\;s^{-1}$. Finally, the presence of $Ca^{2+}$ and $Mg^{2+}$ enhanced the PD-cP activity, with $Mg^{2+}$ 1.4-fold more effective than $Ca^{2+}$.

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