Production and Purification of Acid and Alkaline Phytase enzyme from Native Aspergillus Niger Fungi, Recombinant Escherichia Coli and Bacillus Subtilis Bacterias Aiming a Beneficial Combination for Poultry Diet

In order to increase the availability of phosphorus and other minerals in the diet of birds and monogastrics and reduce environmental pollution this reasearch was aimed to produce phytase. This was designed to produce a suitable enzyme for digestion of phytate in the gastrointestinal tract of birds. For this propose, Aspergillus niger phytase was produced on PSM media and Escherichia coli and Bacillus subtilis phytases with using recombinant protein technology. The enzymes were purified by Q-Sepharose and Ni-NTA chromatography column and, using SDS-PAGE, their molecular weights were estimated 42 kDa, 44 kDa and 66 kDa for PhyC) Alkaline Phytase ( , appA (Escherichia coli Phytase) and PhyA) Acid Phytase ( respectively. The phytase activity for phyC, appA and phyA were determined 54.81, 35.49 and 29.33 U/ml, respectively. Comapring their kinetics parameters, it was shown that phyC enzyme can be active in 90 0C and had thermostability while appA enzyme had best rang of activity in pH of 4-4.5 and efficient enzyme in gastrointestinal tract and appA had maximum affinity to substrate by km 0.09±0.03. According to the results, a suitable combination of these enzymes could be efficient enough to hydrolyse phytate in gastrointestinal tract of birds.