Purification of Glutamate Dehydrogenase from Liver of Rainbow Trout (Oncorhynchus Mykiss)

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Ertik O., Yanardağ R.

2nd International Congress on Medical Sciences and Biotechnology, Uşak, Türkiye, 1 - 03 Ekim 2020, ss.300, (Özet Bildiri)

• Yayın Türü: Bildiri / Özet Bildiri
• Basıldığı Şehir: Uşak
• Basıldığı Ülke: Türkiye
• Sayfa Sayıları: ss.300
• İstanbul Üniversitesi-Cerrahpaşa Adresli: Evet

Özet

Glutamate dehydrogenase (GDH; EC 1.4.1.2-4.) is found in all organisms including bacteria, fungi, plants, fishes and mammals. GDH is a hexameric enzyme that catalyzes the reversible conversion of glutamate to α-ketoglutarate and ammonia while reducing NAD(P)+ to NAD(P)H. In this study, GDH was purified from liver of Rainbow trout (Oncorhynchus mykiss) with a single step using 2',5' ADP Sepharose 4B affinity chromatography. The GDH was purified 171-fold, giving a specific activity of 5.83 U/mg proteins. Optimal conditions for activity of the purified enzyme are as follows: optimal pH 8.00, optimal ionic strength 100 mM, optimal temperature 40ºC. The apparent Michaelis constant values of NAD+ , NADP+ , NADH, NADPH, L-glutamate, α-ketoglutarate and NH4 + were found to be 1.86, 2.17, 2.98, 6.25, 15.15, 4.82 mM and 1.73 M respectively, while the Vmax values are 0.11, 0.05, 0.44, 0.43, 0.12, 0.42 and 1.79 U/mL respectively.