Purification of Glucose 6-Phosphate Dehydrogenase From Goose Erythrocytes and Kinetic Properties

BEYDEMİR Ş., Yilmaz H., Çiftçi M., Bakan E., KÜFREVİOĞLU Ö. İ.

Turkish Journal of Veterinary and Animal Sciences, vol.27, no.5, pp.1179-1185, 2003 (SCI-Expanded) identifier

  • Publication Type: Article / Article
  • Volume: 27 Issue: 5
  • Publication Date: 2003
  • Journal Name: Turkish Journal of Veterinary and Animal Sciences
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Page Numbers: pp.1179-1185
  • Keywords: Erythrocyte, Glucose 6-phosphate dehydrogenase, Goose, Purification
  • Bilecik Şeyh Edebali University Affiliated: Yes

Abstract

Glucose 6-phosphate dehydrogenase (G6PD) was purified from goose erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of 3 steps: hemolysate preparation, ammonium sulfate precipitation, and 2′, 5′-ADP Sepharose 4B affinity gel chromatography. Thanks to the 3 consecutive procedures, the enzyme, having a specific activity of 36.2 EU/mg protein, was purified for a yield of 68.79% and 3892 folds; to ascertain enzyme purity, SDS-PAGE was performed. Optimal pH, stable pH, optimal temperature, molecular weight, and Km and Vmax values for NADP+ and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, Ki values and inhibition type were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP and NADPH. These materials inhibited the enzyme in a noncompetitive manner.