Purification of glucose 6-phosphate dehydrogenase from Buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties.
Glucose 6-phosphate dehydrogenase (G6PD) was purified from buffalo (Bubalus bubalis) erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of two steps: hemolysate preparation and 2('),5(')-ADP-Sepharose 4B affinity gel chromatography. Thanks to the two consecutive procedures, the enzyme, having a specific activity of 69.7EU/mg proteins, was purified 650-fold with a yield of 31%. Optimal pH, stable pH, optimal temperature, molecular weight, and K(M) and V(max) values for NADP(+) and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K(i) values and the type of inhibition were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP, NADPH, and NADH.
Ciftçi
Mehmet
M
Arts and Science Faculty, Department of Chemistry, Atatürk University, Erzurum, Turkey. ciftcim@atauni.edu.tr
Beydemir
Sükrü
S
Yilmaz
Hayrullah
H
Altikat
Sayit
S
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive; D002020:Buffaloes / Q000097:blood*; D002846:Chromatography, Affinity; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology*; D019298:Glucose-6-Phosphate / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000097:blood* / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism / Q000494:pharmacology; D013696:Temperature
D004791:Enzyme Inhibitors; D009249:NADP; D019298:Glucose-6-Phosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D005954:Glucosephosphate Dehydrogenase