Purification and characterization of glucose-6-phosphate dehydrogenase from rat small intestine.
Glucose-6-phosphate dehydrogenase (G6PD) was purified from rat small intestine with 19.2% yield and had a specific activity of 53.8 units per miligram protein. The pH optimum was determined to be 8.1. The purified rat small intestinal G6PD gave one activity, one protein band on native PAGE. The observation of one band on SDS/PAGE with an Mr of 48 kDa and a specific activity lower than expected may suggest the proteolytically affected enzyme or different form of G6PD in the rat small intestine. The activation energy, activation enthalpy, Q10, and optimum temperature from Arrhenius plot for the rat small intestinal G6PD were found to be 8.52 kcal/mol, 7.90 kcal/mol, 1.59, and 38 degrees C, respectively. The Km values for G6P and NADP+ were 70.1 +/- 20.8 and 23.2 +/- 7.6 microM, respectively. Double-reciprocal plots of 1/Vm versus 1/G6P (at constant [NADP+]) and of 1/Vm versus 1/NADP+ at constant [G6P]) intersected at the same point on the 1/Vm axis to give Vm = 53.8 U/mg protein.
Danişan
Ali
A
Department of Biochemistry, Faculty of Medicine, Hacettepe University, 06100 Ankara, Turkey.
Ceyhan
Deniz
D
Oğüş
I Hamdi
IH
Ozer
Nazmi
N
D000818:Animals; D004789:Enzyme Activation; D005260:Female; D019298:Glucose-6-Phosphate / Q000737:chemistry*; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007421:Intestine, Small / Q000201:enzymology*; D008970:Molecular Weight; D009249:NADP / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature; D013816:Thermodynamics
D009249:NADP; D019298:Glucose-6-Phosphate; D005954:Glucosephosphate Dehydrogenase