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Journal Title Abbrev.
J. Elem.
ISSN – 1644-2296
DOI: 10.5601

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Inhibitory effects of Pb2+, Fe2+, Cd2+ and Co2+ on carbonic anhydrase enzyme from muscle of the Kangal fish (Garra rufa)

Issue: 3/2024

Recevied: December 7, 2023

Accepted: September 4, 2024

Published: September 8, 2024

Authors:

U. Kocyigit, I. Gülçin

Categories: Fisheries and animal bioengineering

DOI: 10.5601/jelem.2023.28.4.3215

Abstract:

Kangal fish (Garra rufa) is known as “doctor fish” and lives in Kangal Spring in Sivas, Turkiye. In this study, carbonic anhydrase (CA) was purified and characterized for the first time from the muscle tissues of Kangal fish (Garra rufa). To this end, CA was purified using by Sepharose-4B-L-Tyrosine-sulfanilamide affinity column (STAC) with a specific activity of 34.36 EU.mg−1, a yield of 17.98 % and 201.0 purification fold. To control of the CA enzyme purity, SDS-PAGE was performed and a single band was found. The Michaelis constant (Km) and maximum velocity (Vmax) were determined for CA. Also, p-nitrophenylacetate (PNA) was used as CA substrate. Also, 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 1.0 M (Tris-SO4), optimum pH was calculated 9.0 (Tris-SO4) and stable pH was found 8.5 (phosphate buffer) for the CA from the muscle tissues were found. Furthermore, activation enthalpy (ΔH), the activation energy (Ea), optimum temperature and Q10 values were obtained from Arrhenius plot of CA from Garra rufa muscle tissue as 6.70 kcal mol-1, 7.32 Kcal mol-1, 35.0 °C, 1.37. Kcat and V0 values of CA from Garra rufa muscle CA were calculated as 19.21 s-1 and 1.8x104 mM s-1, respectively. Finally, Ki values of some heavy metal ions (Co2+, Pb2+, Cd2+, and Fe2+) on Kangal fish muscle CA were calculated in range of 0.25-26.09 mM using the esterase activity assay.

Citation:

JELEM / HARVARD

quote-mark
Kocyigit, U. and Gülçin, I. (2024) 'Inhibitory effects of Pb2+, Fe2+, Cd2+ and Co2+ on carbonic anhydrase enzyme from muscle of the Kangal fish (Garra rufa) ', Journal of Elementology, 29(3), 693-712, available: https://doi.org/10.5601/jelem.2023.28.4.3215

MDPI

quote-mark
Kocyigit, U.; Gülçin, I. Inhibitory effects of Pb2+, Fe2+, Cd2+ and Co2+ on carbonic anhydrase enzyme from muscle of the Kangal fish (Garra rufa) . J. Elem. 2024, 29, 3, 693-712. https://doi.org/10.5601/jelem.2023.28.4.3215

AMA

quote-mark
Kocyigit U, Gülçin I. Inhibitory effects of Pb2+, Fe2+, Cd2+ and Co2+ on carbonic anhydrase enzyme from muscle of the Kangal fish (Garra rufa). J. Elem. 2024;29(3): 693-712. https://doi.org/10.5601/jelem.2023.28.4.3215

CHICAGO

quote-mark
Kocyigit Umit M., Gülçin Ilhami. 2024. "Inhibitory effects of Pb2+, Fe2+, Cd2+ and Co2+ on carbonic anhydrase enzyme from muscle of the Kangal fish (Garra rufa) " J. Elem. 29, no.3: 693-712. https://doi.org/10.5601/jelem.2023.28.4.3215



Keywords:

Garra rufa, enzyme purification, Kangal fish, enzyme inhibition, carbonic anhydrase

About issue:

29.3.2024
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