Journal of Applied Biological Sciences

Yazarlar: Ersin HOPA, Selma SİNAN, Emre SEVİNDİK

Konular:-

Anahtar Kelimeler:G6PD,İnhibition,Purification,Pesticide

Özet: This study aims to analyze the purification of Glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase EC 1.1.1.49; G6PD) enzyme, which is important for metabolism, from human erythrocytes and to analyze the effects of some pesticides on enzymes. The purification process was performed in two steps being ammonium sulfate precipitation and 2’, 5’ ADP-Sepharose 4B affinity chromatography. It is identified via sodium dodecyl sulphate polyacrylamide gel (DSD-PAGE) electrophoresis that G6PD enzyme’s molecular weight is about 59000 daltons. Km values for G6PD enzyme’s NADP+ and glucose-6-phosphate (G6P) substrates are respectively 0.14mM, 0.22mM; and, Vmax values are respectively 2.76 U/mg and 1.94 U/mg. Enzyme activities were spectrophotometrically measured on 340 nm according to Beutler method. Pesticides analyzed in this study are deltamethrin and diazinon. After their specific concentration in IC50 studies, deltamethrin and diazinon substances caused turbidity in quartz vessel. Since reliable absorbance values cannot be obtained in such condition, concentration amounts of substances applied to enzyme did not reduce the activity under 50%. Because of this reason, inhibition rates could not be identified. However, it is identified that all of the pesticides show inhibition effect on enzyme activity.

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