CHARACTERIZATION AND PURIFICATION OF THE INTRACELLULAR PHYTASE FROM OCHROBACTRUM ANTHROPI

Abstract

Ochrobactrum anthropi was incubated in fermentation medium including Na-phytate as substrate. During incubation period, the maximum enzyme secretion was determined at 38th, enzyme activity calculated as 69.80 U/mL. Thus, intracellular phytase was purified from the bacterial cells at 38th hour of fermentation. Molecular masses of intracellular phytase from Ochrobactrum anthropi were estimated to be 22 kDa and 13 kDa by SDS-PAGE. Optimal pH and temperature of the phytase for maximum enzyme activity were also recorded as 5.4 and 40 degrees C, respectively. Stability of the intracellular enzyme was revealed to maintain at 90 degrees C for 30 min and its activity increased to 117%. Chemical agents such as NaCl, MnSO4, MgSO4, glycerol, EDTA, acetic acid and 2-mercaptoethanol were detected to cause an increase while SDS, CoCl2, CuSO4, CuCl2, HgCl2, NiCl2, ZnSO4, FeSO4 and FeCl2 showed an inhibition effect on phytase activity depending on the concentration. Intracellular phytase was determined to show a high specificity against other phosphorylated substrates.