Purification of glucose‐6‐phosphate dehydrogenase from rat rat ( Rattus norvegicus Rattus norvegicus ) erythrocytes erythrocytes and inhibition effects of some metal ions on enzyme activity

摘要

Abstract Glucose6phosphate dehydrogenase (G6PD) is the first enzyme on which the pentose phosphate pathway was checked. In this study, purification of a G6PD enzyme was carried out by using rat erythrocytes with a specific activity of 13.7燛U/mg and a yield of 67.7 and 155.6fold by using 22,52ADP Sepharose4B affinity column chromatography. For the purpose of identifying the purity of enzyme and molecular mass of the subunit, a sodium dodecyl sulfatepolyacrylamide gel electrophoresis was carried out. The molecular mass of subunit was calculated 56.5爇Da approximately. Then, an investigation was carried out regarding the inhibitory effects caused by various metal ions (Fe2+ , Pb2+ , Cd2+ , Ag+ , and Zn2+ ) on G6PD enzyme activities, as per Beutler method at 340爊m underin vitro conditions. LineweaverBurk diagrams were used for estimation of the IC50 andK i values for the metals.K i values for Pb+2 , Cd+2 , Ag+ , and Zn+2 were 113.3, 215.2, 19.4, and 474.7牸M, respectively.