Purification and characterization of lactate dehydrogenase from the heart ventricles of river buffalo (Bubalus bubalis)

The present study describes purification and characterization of lactate dehydrogenase (LDH) from the heart ventricles of river buffalo (Bubalus bubalis). Heart specific isozyme of LDH has been purified to apparent homogeneity on SDS-PAGE using ion-exchange column chromatography, selective precipitation in the presence of ammonium sulfate and hydrophobic-interaction chromatography. The enzyme was purified up to 48 fold with 16% recovery. The maximum activity of purified enzyme was observed at pH 7.0 and it has shown reasonable stability at a broad range of temperature with maximum activity at 30 degree C. The Km value with pyruvate is 41 muM, it has only 18% activity with lactate as compared to its activity with pyruvate at pH 7.0. The molecular weight of a subunit of enzyme is 36416.5 plus minus 2 Da as determined by MALDI-TOF analysis.