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

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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.

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Authors and Affiliations

  1. Faculty of Science, Chemistry Department, Biochemistry Division, Erciyes University, 38039, Kayseri, Turkey

    Burcu Somtürk & Nalan Özdemir

  2. Faculty of Science, Chemistry Department, Biochemistry Division, Atatürk University, Erzurum, Turkey

    Ramazan Kalın

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  1. Burcu Somtürk
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Correspondence to Nalan Özdemir.

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Somtürk, B., Kalın, R. & Özdemir, N. Purification of Peroxidase from Red Cabbage (Brassica oleracea var. capitata f. rubra) by Affinity Chromatography. Appl Biochem Biotechnol 173, 1815–1828 (2014). https://doi.org/10.1007/s12010-014-0968-1

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