6-Phosphogluconate dehydrogenase: Purification, characterization and kinetic properties from rat erythrocytes

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

Turkish Journal of Veterinary and Animal Sciences, vol.28, no.4, pp.707-714, 2004 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 28 Issue: 4
  • Publication Date: 2004
  • Journal Name: Turkish Journal of Veterinary and Animal Sciences
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Page Numbers: pp.707-714
  • Keywords: 6PGD, Erythrocyte, Kinetic properties, Purification, Rat
  • Anadolu University Affiliated: Yes


In this paper, a simple and rapid method for the purification of 6-phosphogluconate dehydrogenase from rat erythrocytes together with an analysis of the kinetic behavior and some properties of the enzyme are considered. The purification steps comprised high-speed centrifugation, 20-50% ammonium sulfate precipitation and 2′, 5′-ADP Sepharose 4B affinity gel chromatography. The yield was 78.4% and the specific enzyme activity was 5.15 EU/mg proteins. The molecular mass of the subunit was estimated to be 59,566 Da by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and native enzyme was found to be 111,000 Da by gel filtration column chromatography. The enzyme had an optimal pH at 7.0 and stable pH at 8.0 in 1 M Tris-HCI buffer, and optimal temperature at 45°C. KM and Vmax for NADP+ and 6-PGA as substrates were also determined. The inhibitor effects of ATP, NADPH and NADH were also examined, and Ki values and the types of inhibition were determined by means of a Lineweaver-Burk graph obtained for them.