Purification of thermo and acid tolerant extracellular phytase from a new soil isolate of Amycolatopsis vancoresmycina S-12

Abstract

Amycolatopsis vancoresmycina S-12, a member of soil Actinomycetes, was explored regarding its extracellular phytase. The enzyme was recovered from the culture broth by ammonium sulfate fractionation and purified to near homogeneity following dialysis, DEAE Sepharose and Sphadex G-100 column. The enzyme was purified 5.51 fold with a final yield of 14.9%. It was characterized by SDS PAGE, Zymogram, Native PAGE, 2D PAGE as monomeric protein of 53 kDa. and pI 5.4. Peptide mass fingerprinting of MALDI-TOF MS result indicates its phytase nature. Optimum enzyme activity was at 37 °C and at pH 5.4. Optimum stability was at 60 °C and pH 8.0. The enzyme activity is stimulated in presence of Mg2+. The plant growth promotion experiment shows a significant effect, with an increase of 10% root and 25% of shoot length, resulting in a 2.2 fold increase in biomass.