Purification and Kinetic Properties of Glucose-6-Phosphate Dehydrogenase from Goat(Capra aegagrus hicrus)Erythrocytes

摘要

The aim of this study was to purify glucose-6-phosphate dehydrogenase(G6PD)from domestic goat(Capra aegagrus hicrus)erythrocytes and to investigate its some in vitro kinetic properties.Blood was taken on ACD-A solution and erythrocytes were purified,and then they were hemolysated.In the study DEAE-cellulose as anion and CM-Sephadex as cation exchange column chromatographies and ammonium sulphate precipitation and dialysis methods were used.G6PD was purified 45000 fold in a yield of 23 %.Molecular weight was found 210000 by Sephadex G-200.Four subunits of this enzyme were determined and their molecular weights were found 52000.K_m and V_(max)values for G6PD were determined as 3.34 x 10~(-4)M and 642 mum/min.In addition we found that levels of Mg~(2+)from 1 x 10~(-3)M to 6 x 10~(-3)M are increasing activity of G6PD and 10~(-4)M dehydroepiandrosterone is preventing whole activity of G6PD.