Purification and biochemical characterization of a novel carbonic anhydrase II from erythrocytes of camel (Camelusdromedarius)

Abdelbasset Chafik; Abdelkhalid Essamadi; Safinur Yildirim Çelik; Ahmet Mavi

doi:10.1016/j.bbrc.2023.07.055

Purification and biochemical characterization of a novel carbonic anhydrase II from erythrocytes of camel (Camelusdromedarius)

Biochem Biophys Res Commun. 2023 Oct 8:676:171-181. doi: 10.1016/j.bbrc.2023.07.055. Epub 2023 Jul 26.

Authors

Abdelbasset Chafik¹, Abdelkhalid Essamadi², Safinur Yildirim Çelik³, Ahmet Mavi⁴

Affiliations

¹ Higher School of Technology of El Kelâa des Sraghna, Cadi Ayyad University, Beni Mellal Road Km 8, BP 104, El Kelâa des Sraghna, 43000, Morocco; Laboratory of Biochemistry, Neurosciences, Natural Resources and Environment, Faculty of Sciences and Techniques, Hassan First University, Settat, 26000, Morocco; Bioresources and Food Safety Laboratory, Faculty of Sciences and Techniques, Cadi Ayyad University, Boulevard Abdelkrim Khattabi, BP 549, Marrakech, 40000, Morocco. Electronic address: a.chafik@uca.ma.
² Laboratory of Biochemistry, Neurosciences, Natural Resources and Environment, Faculty of Sciences and Techniques, Hassan First University, Settat, 26000, Morocco.
³ Department of Nutrition and Dietetics, Faculty of Health Sciences, Atatürk University, 25240, Erzurum, Turkey.
⁴ Chemistry Laboratory, Department of Mathematics and Science Education, Kazim Karabekir Education Faculty, Atatürk University, 25240, Erzurum, Turkey; Department of Nanoscience & Nanoengineering, Graduate School of Natural & Applied Science, Atatürk University, Erzurum, Turkey.

PMID: 37517220
DOI: 10.1016/j.bbrc.2023.07.055

Abstract

A novel carbonic anhydrase II (CA II) from erythrocytes of camel (Camelus dromedarius) was purified to homogeneity using affinity chromatography and biochemically characterized. Specific activity of 140.88 U/mg was obtained with 745.17-fold purification and 25.37% yield. The enzyme was a monomer with a lower molecular weight (25 kDa) and lower Zn content (0.50 mol of Zn per mol of protein). The enzyme showed higher optimum temperature (70 °C) and pH (pH 9.0), moreover, it was stable at higher temperatures and strongly alkaline pH as judged by thermodynamic parameters (E_a, k_d, E_d, t_1/2, D-value, Z-value, ΔH, ΔG and ΔS). The enzyme was inhibited by cations (Al³⁺, Ca²⁺, Cd²⁺, Co²⁺, Cr³⁺, Cu²⁺, Fe³⁺, Ni²⁺, Mg²⁺ and Zn²⁺) as well as by anions (Br^‾, CH₃COO^‾, ClO₄^‾, CN^‾, F^‾, HCO₃^‾, I^‾, N₃^‾, NO₃^‾ and SCN^‾), some anions (C₆H₅O₇^3-, CO₃^2-, SeO₃^‾ and SO₄^2-) does not affect enzyme activity. Effect of various chemicals on enzyme activity was also investigated. K_m, V_max, k_cat and k_cat/K_m values for 4-NPA were found to be 1.74 mM, 0.0093 U/mL, 0,0039 s^-1 and 0,0023 s^-1 mM^-1, respectively. With these interesting biochemical properties, camel CA II represents promising candidate for harsh industrial applications, in particular, for a successful biomimetic CO₂ sequestration process.

Keywords: Biochemical characterization; Camel; Camelus dromedarius; Carbonic anhydrase II; Purification.