Purification of glucose 6-phosphate dehydrogenase from buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties

Çiftçi M., BEYDEMİR Ş., Yilmaz H., Altikat S.

Protein Expression and Purification, vol.29, no.2, pp.304-310, 2003 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 29 Issue: 2
  • Publication Date: 2003
  • Doi Number: 10.1016/s1046-5928(03)00073-1
  • Journal Name: Protein Expression and Purification
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.304-310
  • Keywords: Buffalo, Erythrocyte, Glucose 6-phosphate dehydrogenase, Purification
  • Bilecik Şeyh Edebali University Affiliated: Yes

Abstract

Glucose 6-phosphate dehydrogenase (G6PD) was purified from buffalo (Bubalus bubalis) erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of two steps: hemolysate preparation and 2′,5′-ADP-Sepharose 4B affinity gel chromatography. Thanks to the two consecutive procedures, the enzyme, having a specific activity of 69.7 EU/mg proteins, was purified 650-fold with a yield of 31%. 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 the type of inhibition were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP, NADPH, and NADH. © 2003 Elsevier Science (USA). All rights reserved.