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Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leavesÇoban, T. Abdül KadirÇifti, MehmetKüfrevioglu, O. IrfanChromatography, Ion ExchangeElectrophoresis, Polyacrylamide GelGlucosephosphate DehydrogenaseKineticsMolecular WeightPetroselinumPlant LeavesThermodynamicsPetroselinum crispumglucose 6 phosphate dehydrogenasearticleenzymologyion exchange chromatographyisolation and purificationmetabolismparsleyplant leafpolyacrylamide gel electrophoresisIn this study, glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps: preparation of homogenate, ammonium sulfate fractionation, and DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 8.79% and had a specific activity of 2.146 U (mg protein)-1. The overall purification was about 58-fold. Temperature of +4°C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method, at 340 nm. In order to control the purification of enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 77.6 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a molecular weight of 79.3 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found to be 6.0, 8.0, and 60°C, respectively. Moreover, KM and Vmax values for NADP+ and G6-P at optimum pH and 25°C were determined by means of Line-weaver-Burk graphs. Additionally, effects of streptomycin sulfate and tetracycline antibiotics were investigated for the enzyme activity of glucose-6-phosphate dehydrogenase in vitro.2002Articlehttps://rdlopen.rawdatalibrary.net/r/ebyu/16083https://doi.org/10.1081/pb-1200041291082-6068enPreparative Biochemistry and Biotechnologyinfo:eu-repo/semantics/openAccessinfo:eu-repo/semantics/articlehttps://ebyu-rdlopen.rawdatalibrary.net/tr/items/ebyu/16083