Purification of glucose 6-phosphate dehydrogenase from goose erythrocytes and kinetic properties

BEYDEMIR S., YILMAZ H., CIFTCI M., Bakan E., Kufrevioglu O.

TURKISH JOURNAL OF VETERINARY & ANIMAL SCIENCES, cilt.27, sa.5, ss.1179-1185, 2003 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 27 Sayı: 5
  • Basım Tarihi: 2003
  • Dergi Adı: TURKISH JOURNAL OF VETERINARY & ANIMAL SCIENCES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.1179-1185
  • Anahtar Kelimeler: goose, purification, glucose 6-phosphate dehydrogenase, erythrocyte, ENZYME-ACTIVITY, IN-VITRO, PROTEINS, VIVO
  • Atatürk Üniversitesi Adresli: Evet

Özet

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 K-m and V-max values for NADP(+) and glucose 5-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K-i 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.