Purification, characterisation and kinetic properties of 6-phosphogluconate dehydrogenase enzyme (E.C. 1.1.1.44) from the Van Cat erythrocyte

Abstract

In this study, 6-phosphogluconate dehydrogenase enzyme (6PGD; E.C.1.1.1.44) was purified from erythrocyte the Van Cat by affinity chromatography. The inhibitor effect of Netilmicin sulphate, Ampicillin and Amoxicillin were also examined. Enzyme activity was measured by spectrophotometer according to Beutler method at 340 nm. Then the optimum pH and optimum temperature of the enzyme were determined to be 8.0 and 50 ° C, respectively. In order to control the purification of enzyme, sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used. The purification rate for erythrocyte 6PGD was found as 1787. KM and Vmax were also determined for NADP+ and 6PGA as substrates. The inhibitor effect of Netilmicin sulphate, Ampicillin and Amoxicillin were also examined and Ki values and the types of inhibition were determined by the Lineweaver-Burk graph.