Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes. Investigation of some kinetic properties

Ylmaz H., Ciftci M., BEYDEMİR Ş., Bakan E.

Preparative Biochemistry and Biotechnology, vol.32, no.3, pp.287-301, 2002 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 32 Issue: 3
  • Publication Date: 2002
  • Doi Number: 10.1081/pb-120013475
  • Journal Name: Preparative Biochemistry and Biotechnology
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.287-301
  • Bilecik Şeyh Edebali University Affiliated: Yes

Abstract

Glucose 6-phosphate dehydrogenase (G6PD) was purified from chicken erythrocytes, and some characteristics of the enzyme were investigated. The purification procedure was composed of three steps: hemolysate preparation, ammonium sulfate precipitation, and 2′,5′-ADP Sepharose 4B affinity gel chromatography. Thanks to the three consecutive procedures, the enzyme, having the specific activity of 20.862 EU/mg proteins, was purified with a yield of 54.68% and 9,150-fold. Optimal pH, stable pH, optimal temperature, molecular weight, and KM and Vmax values for NADP+ and glucose 6- phosphate (G6-P) were also determined for the enzyme. In addition, Ki values and the type of inhibition were determined by means of Line-Weaver-Burk graphs obtained for such inhibitors as ATP, ADP, NADH, and NADPH.