Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity |
| |
| Affiliation: | 1. Atatürk University, Faculty of Sciences, Department of Chemistry, Erzurum, Turkey;2. King Saud University, College of Science, Department of Zoology, Riyadh, Saudi Arabia;1. Department of Chemistry, Faculty of Science, Ataturk University, 25240 Erzurum, Turkey;2. Department of Biotechnology, Faculty of Science, Bartin University, 74100 Bartin, Turkey;1. Laboratory of Biochemistry and Neuroscience, Team of Applied Biochemistry and Toxicology, Faculty of Science and Technology, University Hassan First, 577 Settat, Morocco;2. College of Education, Bayburt University, 69000 Bayburt, Turkey;3. Chemistry Education, Kazim Karabekir Education Faculty, Atatürk University, 25240 Erzurum, Turkey;1. Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel;2. Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel;1. Department of Neurosurgery, Peking University People''s Hospital, No. 11, Xizhimen South Street, Xicheng District, Beijing, 100044, China;2. Department of Neurosurgery, Beijing Beiya Orthopedic Hospital, No. 20, Haotian North Street, Fangshan District, Beijing, 102445, China;3. Sanmenxia Key Laboratory of Neurotumor Diagnosis and Treatment, Department of Neurosurgery, Sanmenxia Central Hospital, 472000, Sanmenxia, Henan, China;4. College of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, 471000, China;1. Department of Pharmacology, School of Medicine, Keio University, 160-8582, Japan;2. Keio Advanced Institute for Water Biology and Medicine, Japan;1. Department of Chemistry, Faculty of Science, Akdeniz University, 07058, Antalya, Turkey;2. Central Research and Application Laboratory, Ağrı İbrahim Çeçen University, 04100, Ağrı, Turkey;3. Department of Chemistry, Faculty of Science, Atatürk University, 25240, Erzurum, Turkey;4. Department of Engineering Sciences, Faculty of Engineering, İzmir Katip Çelebi University, 35620, İzmir, Turkey;5. Faculty of Pharmacy, Ağrı İbrahim Çeçen University, 04100, Ağrı, Turkey |
| |
| Abstract: | In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77 EU/mg and a yield of 35.5% using Sepharose-4B-l-tyrosine- sulphanilamide affinity column chromatography. For determining the enzyme purity and subunit molecular mass, sodiumdodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The molecular mass of subunit was found approximately 29.71 kDa. The optimum temperature, activation energy (Ea), activation enthalpy (ΔH) and Q10 values were obtained from Arrhenius plot. Km and Vmax values for p-nitrophenyl acetate of the purified enzyme were calculated from Lineweaver-Burk graphs. In addition, the inhibitory effects of different heavy metal ions (Fe2+, Pb2+, Co2+, Ag+ and Cu2+) on Black Sea trout kidney tissue CA enzyme activities were investigated by using esterase method under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC50) were obtained. Finally Ki values and inhibition types were calculated from Lineweaver-Burk graphs. |
| |
| Keywords: | Black Sea trout Enzyme purification Enzyme characterization Carbonic anhydrase Heavy metal |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
