Investigation of purified carbonic anhydrase activity from the gills of pelagic and benthic fish

In this study, carbonic anhydrase (CA) was purified and characterized from the gills of anchovy fish living in the pelagic region and whiting living in the benthic region in marine environment. Carbonic anhydrase from the whiting gill was purified 14 fold and %19.5 yield using Sepharose-4B-L tyrosine-sulfanilamide affinity column. Also, carbonic anhydrase from the anchovy gill was purified 17 fold and %9.5 yield using same column. The specific activity of whiting and anchovy fish were determined as 126.4 EU / mg protein and 1,000.0 EU / mg protein, respectively. SDS-PAG Electrophoresis showed that the carbonic anhydrases purified from the gills of both fish had single protein bands with a subunit molecular mass of approximately 29 kDa. In the presence of p-nitrophenyl acetate substratum of gills, esterase activities were found to be highest at pH 8.0 and 40 ° C. The values of Km and Vmax of carbonic anhydrase from the gills of whiting and anchovy fish were calculated by Lineweaver-Burk graph in the presence of p-nitrophenol acetate substrate and Km values were determined as 0.08 mM and 0.01 mM respectively, Vmax value was 1x107 M / min and 2.5x106 M / min, respectively. It was determined that the CA obtained from the gills of whiting and anchovy fish had an IC50 value of 6.0 μM to 4.0 μM against the sulfanilamide inhibitör, respectively and 2.0 μM to 2.0 μM, against the acetazolamide inhibitör.