Kuzu M.,University of Agr Ibrahim Cecen |
Aslan A.,University of Agr Ibrahim Cecen |
Ahmed I.,Karlsruhe Institute of Technology |
Comakli V.,University of Agr Ibrahim Cecen |
And 2 more authors.
Fish Physiology and Biochemistry | Year: 2015
Glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) are metabolically quite important enzymes. Within this study, these two enzymes were purified for the first time from the gills of Lake Van fish. In the purifying process, ammonium sulfate precipitation and 2′,5′-ADP Sepharose 4B affinity column chromatography techniques for glucose-6-phosphate dehydrogenase, temperature degradation and 2′,5′-ADP Sepharose 4B affinity column chromatography for glutathione reductase enzyme were used. The control of the enzyme purity and determination of molecular weight were done with sodium dodecyl sulfate polyacrylamide gel electrophoresis. KM and Vmax values were determined with Lineweaver–Burk plot. Besides, the effects of some chalcone derivatives on the purified enzymes were analyzed. For the ones showing inhibition effect, % activity–[I] figures were drawn and IC50 values were determined. Ki value was calculated by using Cheng–Prusoff equation. © 2015 Springer Science+Business Media Dordrecht