Purification of Peroxidase from Red Cabbage (Brassica oleracea var. capitata f. rubra) by Affinity Chromatography


SOMTURK B. , Kalin R., ÖZDEMİR N.

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, vol.173, no.7, pp.1815-1828, 2014 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 173 Issue: 7
  • Publication Date: 2014
  • Doi Number: 10.1007/s12010-014-0968-1
  • Title of Journal : APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
  • Page Numbers: pp.1815-1828

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 A degrees 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 A +/- 0.05 mM, respectively, and 4-amino benzohydrazide showed noncompetitive inhibition.