Extracellular phytase from Aspergillus niger CFR 335: purification and characterization.

Abstract

Phytase, that is extensively used as a feed additive is capable of hydrolyzing phytic acid, an antinutrient found in about 60-80% of all the plant commodities. This enzyme improves the bioavailability of essential minerals such as Ca(2+), Mg(2+), P, Zn(2+), Fe(3+), that are bound to phytic acid. An extracellular phytase from a local fungal isolate, Aspergillus niger CFR 335 was purified to homogeneity through a three-step column chromatography using DEAE-Sephadex anion exchanger. An active fraction of the enzyme was obtained with NaCl gradient of 2.5M in DEAE Sephadex column. The enzyme was purified up to 16 fold with a yield of 28.5%. Substrate specificity studies revealed a highest specific activity of 32.6 ± 3.1 U/mg for sodium phytate with the Km value of 0.08 ± 0.1mM. The molecular weight of the enzyme was 66kDa with an optimum temperature of 30°C and pH4.5. Up to 80% of the activity was retained even after storing the enzyme for 6months at 4°C.