Purification, characterisation and kinetic properties of 6-phosphogluconate dehydrogenase enzyme (E.C. 1.1.1.44) from the Van Cat erythrocyte Van Kedisi Eritrositi'nden 6-Fosfoglukonat Dehidrogenaz Enzimi (E.C. 1.1.1.44)'nin Saflaştirilmasi, Karakterizasyonu ve Kinetik Özellikleri

KILIÇ S., Karataş F.

Ataturk Universitesi Veteriner Bilimleri Dergisi, cilt.12, sa.2, ss.187-194, 2017 (Scopus, TRDizin) identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 12 Sayı: 2
  • Basım Tarihi: 2017
  • Doi Numarası: 10.17094/ataunivbd.347975
  • Dergi Adı: Ataturk Universitesi Veteriner Bilimleri Dergisi
  • Derginin Tarandığı İndeksler: Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.187-194
  • Anahtar Kelimeler: 6-Phosphogluconate dehydrogenase enzyme, Affinity chromatography, Erythrocyte, Kinetic properties, Van Cat
  • Isparta Uygulamalı Bilimler Üniversitesi Adresli: Hayır

Özet

In this study, 6-phosphogluconate dehydrogenase enzyme (6PGD; E.C.1.1.1.44) was purified from erythrocyte the Van Cat by affinity chromatography. The inhibitor effect of Netilmicin sulphate, Ampicillin and Amoxicillin were also examined. Enzyme activity was measured by spectrophotometer according to Beutler method at 340 nm. Then the optimum pH and optimum temperature of the enzyme were determined to be 8.0 and 50 ° C, respectively. In order to control the purification of enzyme, sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used. The purification rate for erythrocyte 6PGD was found as 1787. KM and Vmax were also determined for NADP+ and 6PGA as substrates. The inhibitor effect of Netilmicin sulphate, Ampicillin and Amoxicillin were also examined and Ki values and the types of inhibition were determined by the Lineweaver-Burk graph.