Purification and characterization of peroxidase from cauliflower (Brassica oleracea L. var. botrytis) buds


Koksal E. , GÜLÇİN İ.

PROTEIN AND PEPTIDE LETTERS, cilt.15, ss.320-326, 2008 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 15 Konu: 4
  • Basım Tarihi: 2008
  • Doi Numarası: 10.2174/092986608784246506
  • Dergi Adı: PROTEIN AND PEPTIDE LETTERS
  • Sayfa Sayıları: ss.320-326

Özet

Peroxidases (EC 1.11.1.7; donor: hydrogen peroxide oxidoreductase) are part of a large group of enzymes. In this study, peroxidase, a primer antioxidant enzyme, was purified with 19.3 fold and 0.2% efficiency from cauliflower (Brassica oleracea L.) by ammonium sulphate precipitation, dialysis, CM-Sephadex ion-exchange chromatography and Sephadex G-25 purification steps. The substrate specificity of peroxidase was investigated using 2,2'-azino-bis 3-ethylbenz-thiazoline-6-sulphonic acid) (ABTS), 2-methoxyphenol (guaiacol), 1,2-dihydroxybenzene (catechol), 1,2,3-trihyidroxybenzene (pyrogallol) and 4-methylcatechol. Also, optimum pH, optimum temperature, optimum ionic strength, stable pH, stable temperature, thermal inactivation conditions were determined for guaiacol/H2O2, pyrogallol/H2O2, ABTS/H2O2, catechol/H2O2 and 4-methyl catechol/ H2O2 substrate patterns. The molecular weight (M-w) of this enzyme was found to be 44 kDa by gel filtration chromatography method. Native polyacrylamide gel electrophoresis (PAGE) was performed for isoenzyme determination and a single band was observed. K-m and V-max values were calculated from Lineweaver-Burk graph for each substrate patterns.