Purification and biochemical characterization of catalase that confers protection against hydrogen peroxide induced by stressful desert environment: the Camelus Dromedarius kidney catalase

Abstract

Camel is continually exposed to stressful desert environment that enhances generation of reactive oxygen species, including hydrogen peroxide (H₂O₂). Catalase plays an important role in detoxification of H₂O₂. A highly active catalase from camel kidney was purified to homogeneity, with a specific activity of 1,774,392 U/mg protein, using ion exchange and metal chelate affinity chromatography. The molecular weight of the enzyme was 268 kDa consisting of four identical subunits of 63 kDa. The enzyme showed higher optimum temperature (45 °C) and higher activation energy (4.37 kJ mol⁻¹). The thermodynamic parameters, ΔH, ΔG and ΔS, were determined. The effect of various metal ions and chemicals on enzyme activity was investigated. K_m, V_max, k_cat and k_cat/K_m values for H₂O₂ were found to be 46 mM, 10,715,045 U/mg, 48,265,968 s⁻¹ and 2,966,562 s⁻¹mM⁻¹, respectively. Camel kidney catalase displayed higher affinity efficiency for H₂O₂ and can protect reduced glutathione (GSH) from oxidation by H₂O₂. Sodium azide was found to be a noncompetitive inhibitor of enzyme with K_i and IC₅₀ of 17.88 µM and 20.94 µM, respectively. Camel catalase showed unique biochemical properties. Interestingly, camel catalase can protect molecules (GSH) and organ functions (kidney) from the toxic effects of H₂O₂ induced by stressful desert environment.

Keywords:

• Biochemical characterization
• camel
• catalase
• purification
• stressful desert environment

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

The authors thank Atatürk University (Turkey) and University Hassan First (Morocco) for their financial support.