Purification and characterization of a protease-resistant phytase of Aspergillus oryzae SBS50 whose properties make it exceptionally useful as a feed supplement

An extracellular phytase of Aspergillus oryzae SBS50 was purified to homogeneity using ammonium sulphate precipitation, ion-exchange and gel filtration chromatography. Purified phytase has a monomeric molecular mass of ∼80 kDa exhibiting its optimal activity at pH 5.0 and 50 °C with a T 1/2 of 300 min at 50 °C. Phytase of A. oryzae displayed broad substrate specificity with Vmax and Km values of 58.82 μmol/ml/min and 1.14 mM, respectively, for calcium phytate. Purity and homogeneity of the phytase was confirmed by high performance liquid chromatography and MALDI-TOF analysis revealed the identification of a peptide showing homology with acid phosphatase of Aspergillus oryzae RIB40. Among the inhibitors, 2,3-butanedione and sodium molybdate significantly inhibited the enzyme activity. Phytase of A. oryzae showed protease-resistance and was more stable during storage at 4 °C and −20 °C as compared to room temperature. Among all the feed samples, mustard oil cake was dephytinized more efficiently than other feed samples. These unique properties suggested that the phytase has the potential to be useful as an animal feed supplement.