Author(s)

Tang Jing, Tang Yunming

Corresponding Author

Tang Yunming

Abstract

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 70.07and its specific activity was13.63U/mg. 24.61% of the glutamate dehydrogenase activity was recovered. The enzyme had a relative molecular mass of 330.08KD. The relative molecular mass of the subunit was about 56.04KD. 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 0.084mmol/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 .

Keywords

Porcine brain, glutamate dehydrogenase, isolation and purification, characterization