Purification and characterization of glucose 6-phosphate dehydrogenase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the effects of some metal ions on enzyme activity

Comakli V., AKKEMIK E., Ciftci M., KÜFREVİOĞLU Ö. İ.

TOXICOLOGY AND INDUSTRIAL HEALTH, cilt.31, sa.5, ss.403-411, 2015 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 31 Sayı: 5
  • Basım Tarihi: 2015
  • Doi Numarası: 10.1177/0748233713475514
  • Dergi Adı: TOXICOLOGY AND INDUSTRIAL HEALTH
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.403-411
  • Anahtar Kelimeler: Glucose 6-phosphate dehydrogenase, liver, rainbow trout, metal ion, inhibition, ERYTHROCYTE GLUTATHIONE-REDUCTASE, KINETIC-PROPERTIES, IN-VITRO, DEHYDROGENASE, VIVO
  • Atatürk Üniversitesi Adresli: Evet

Özet

Glucose 6-phosphate dehydrogenase (d-glucose 6-phosphate: NADP(+) oxidoreductase, EC 1.1.1.49; G6PD) is a key enzyme that is localized in all mammal tissues, especially in cytoplasmic sections and that catalyzes the first step of pentose phosphate metabolic pathway. In this study, G6PD enzyme was purified 1444-fold with a yield of 77% from rainbow trout liver using 2,5-ADP-sepharose-4B affinity chromatography. Moreover, a purity check of the enzyme was performed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Some characteristic features like optimal pH, stable pH, optimal temperature and optimal ionic strength were determined for the purified enzyme. In addition to this, in vitro effects of ions like silver nitrate (Ag+), thallium sulphate (TI+), cobalt (II) nitrate (Co2+) and arsenic (V) oxide (As5+) on enzyme activity were researched. Half-maximal inhibitory concentration (IC50) values of Ag+, Co2+ and As5+ metal ions, which showed an inhibitory effect, were found to be 0.0044, 0.084 and 4.058mM, respectively; and their inhibition constants (K-i) were found to be 0.0052 +/- 0.00042, 0.087 +/- 0.015700 and 4.833 +/- 1.753207mM, respectively. Tl+ not exhibited inhibitory effect on the enzyme activity.