Ekrem Koksal and Ilhami Gulcin Pages 320 - 326 ( 7 )
Peroxidases (EC 126.96.36.199; 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 (Mw) 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. Km and Vmax values were calculated from Lineweaver-Burk graph for each substrate patterns.
Cauliflower, Brassica oleracea, peroxidase, enzyme purification, chromatography
Ataturk University, Science and Arts Faculty, Department of Chemistry, TR-25240, Erzurum- Turkey.