Cloning and optimization of phytase enzyme gene expression in Escherichia coli

Authors
Maryam Azadi 1
leila pishkar 2
Ehsan Dehnavi 3
1 Department of Biochemistry Tehran University of Medical Sciences, Islamic Azad University, Tehran, Iran
2 Department of Biology, Islamshahr Branch, Islamic Azad University, Islamshahr, Iran
3 Pharmaceutical Technology Development Center of Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract
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.
Keywords
Phytase 6-phytase
Cloning
Escherichia coli
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Journal of Netshabur University of Medical Sciences
Volume 9, Issue 2 - Serial Number 31
Spring 2021
Pages 118-125