Isolation, Purification and Characterization of Glutamate Dehydrogenase from Porcine Brain
Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweet Potato Engineering Research Center, School of Life Science, Southwest University
摘要：To obtain pure glutamate dehydrogenase from porcine brain and explore its enzymological properties, the glutamate dehydrogenase which showed a single band on SDS-PAGE had been purified from porcine brain by frozen, homogenization, ammonium sulfate precipitation, DEAE-Sepharose chromatography and Superdex-200 chromatography. Its multiple of purification was 70.07 and its specific activity was13.63 U/mg. 24.61% of the glutamate dehydrogenase activity was recovered. The enzyme had a relative molecular mass of 330.08 KD. The relative molecular mass of the subunit was about 56.04 KD. The optimum temperature and pH of this enzyme were 55℃ and 8.2 respectively. The glutamate dehydrogenase displayed excellent stability at temperature below 40℃ and pH 6-8. Its apparent Km towards NADH was 0.084 mmol/L. The enzyme activity could be strongly inhibited when interacting with methanol, ethanol, isopropanol, SDS, oxalic acid, ascorbic acid, Cu2+, Co2+, or Zn2+ and activated when interacting with EDTA.
2019 1st International Biology and Medicine Conference（IBMC 2019）