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

• Published in: Biochemical and biophysical research communications 2023-10, Vol.676, p.171-181
• Main Authors: Chafik, Abdelbasset, Essamadi, Abdelkhalid, Çelik, Safinur Yildirim, Mavi, Ahmet
• Format: Article
• Language: English
• Subjects: Biochemical characterization; Camel; Camelus dromedarius; Carbonic anhydrase II; Purification
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2023.07.055

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 (Ea, kd, Ed, t1/2, 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 (C6H5O73−, CO32−, SeO3‾ and SO42−) does not affect enzyme activity. Effect of various chemicals on enzyme activity was also investigated. Km, Vmax, kcat and kcat/Km 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. •Purification of a novel CA II from camel erythrocytes was carried out.•Camel erythrocytes CA II was biochemically characterized.•CA II showed higher optimum temperature (70 °C) and pH (pH 9.0).•CA II was stable at higher temperatures and strongly alkaline pH.•CA II represents promising candidate for harsh industrial applications.