Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes. investigation of some kinetic properties.
Glucose 6-phosphate dehydrogenase (G6PD) was purified from chicken erythrocytes, and some characteristics of the enzyme were investigated. The purification procedure was composed of three steps: hemolysate preparation, ammonium sulfate precipitation, and 2',5'-ADP Sepharose 4B affinity gel chromatography. Thanks to the three consecutive procedures, the enzyme, having the specific activity of 20.862 EU/mg proteins, was purified with a yield of 54.68% and 9,150-fold. Optimal pH, stable pH, optimal temperature, molecular weight, and KM and Vmax values for NADP+ and glucose 6- phosphate (G6-P) were also determined for the enzyme. In addition, Ki values and the type of inhibition were determined by means of Line-Weaver-Burk graphs obtained for such inhibitors as ATP, ADP, NADH, and NADPH.
Yilmaz
Hayrullah
H
Department of Chemistry, Faculty of Education, Dicle University, Erzurum, Turkey.
Ciftci
Mehmet
M
Beydemir
Sukru
S
Bakan
Ebubekir
E
D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology*; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013696:Temperature
D005954:Glucosephosphate Dehydrogenase