Akademik Veri Yönetim Sistemi
JOURNAL OF ELEMENTOLOGY, sa.3, 2024 (SCI-Expanded)
The Kangal fish (Garra rufa), known as the "doctor fish", lives in the Kangal Spring in Sivas, Turkiye. In this study, carbonic anhydrase (CA) was purified and characterized for the first time from the muscle tissues of the Kangal fish (Garra rufa). To this end, CA was purified using a Sepharose-4B-L-Tyrosine-sulfanilamide affinity column (STAC) with specific activity of 34.36 EU.mg(-1), yield of 17.98 % and 201.0 purification fold. To control the CA enzyme purity, SDS-PAGE was performed and a single band was found. The Michaelis constant (K-m) and maximum velocity (V-max) were determined for CA. Also, p-nitrophenylacetate (PNA) was used as CA substrate. Furthermore, inhibition constants (Ki) and half maximal enzyme inhibitory concentration (IC50) for each metal ion were determined using by Lineweaver-Burk graphs. Additionally, optimum ionic strength was found to be 1.0 M (Tris-SO4), optimum pH was calcu-lated as 9.0 (Tris-SO4) and stable 8.5 pH was found (phosphate buffer) for the CA from the muscle tissues of the fish. Furthermore, activation enthalpy (Delta H), activation energy (Ea), optimum temperature and Q(10) values were obtained from the Arrhenius plot of CA from Garra rufa muscle tissue as 6.70 kcal/mol, 7.32 Kcal mol(-1), 35.0 oC, 1.37, respectively. Kcat and V0values of CA from Garra rufa muscle CA were calculated as 19.21 s(-1) and 1.8x10(4) mM s(-1), respectively. Finally, Ki values of some heavy metal ions (Co2+, Pb2+, Cd2+, and Fe2+) in the Kangal fish muscle CA were calculated in the range of 0.25-26.09 mM using the esterase activity assay.