Cloning and optimization of phytase enzyme gene expression in Escherichia coli

Introduction Phytase is an enzyme that has the ability to break down phytic acid into myoinositol and mineral phosphate, and widely uses as an additive in animal foods. The aim of this study was to achieve a high level of bacterial phytase expression in PET26b expression host. Materials and Methods To generate the recombinant phytase enzyme, the target gene was introduced into the expression vector pET-26b-Phytase, and the recombinant vector was transferred to Escherichia coli BL21 as a host of expression after replication in E. coli DH5α. The SDS-PAGE method was used to isolate recombinant phytase and the amount of produced recombinant protein was investigated. by electrophoresis. Results The bacterial phytase enzyme was successfully expressed in Escherichia coli and the molecular weight of recombinant phytase produced at about 85 kDa was estimated. The optimum pH for recombinant bacterial phytase was estimated at 5.5. Conclusion The results showed that phytase derived from Escherichia coli due to its low production cost and high production of recombinant phytase is a desirable alternative for use in domestic animal feed industry.