title: Purification of peroxidase from red cabbage (Brassica oleracea var. capitata f. rubra) by affinity chromatography.
abstract: Peroxidase was purified in a single step using 4-amino benzohydrazide affinity chromatography from red cabbage (Brassica oleracea var. capitata f. rubra), and some important biochemical characteristics of the purified enzyme were determined. The enzyme, with a specific activity of 3,550 EU/mg protein, was purified 120.6-fold with a yield of 2.9% from the synthesized affinity matrix. The molecular weight of the enzyme was found to be 69.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited maximum activity at pH 7.0 and 30 °C. For guaiacol substrate, the K m and V max values were found as 0.048 mM and 1.46 EU/mL/min, respectively. Additionally, the IC50 and K i values for 4-amino benzohydrazide were calculated to be 1.047 and 0.702±0.05 mM, respectively, and 4-amino benzohydrazide showed noncompetitive inhibition.
authors.affiliation: Faculty of Science, Chemistry Department, Biochemistry Division, Erciyes University, 38039, Kayseri, Turkey.
authors.forename: Burcu
authors.initials: B
authors.lastname: Somtürk
authors.forename: Ramazan
authors.initials: R
authors.lastname: Kalın
authors.forename: Nalan
authors.initials: N
authors.lastname: Özdemir
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology; D001937:Brassica / Q000201:enzymology*; D002846:Chromatography, Affinity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D006139:Guaiacol / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008970:Molecular Weight; D009994:Osmolar Concentration; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000814:Aniline Compounds; D004791:Enzyme Inhibitors; C034343:4-aminobenzhydrazide; D006139:Guaiacol; D006861:Hydrogen Peroxide; D009195:Peroxidase