A Comparative Analysis of Recombinant Expression and Solubility Screening of Two Phytases in E. coli

Ushasree Mrudula Vasudevan, Sumayya Husaiba Beevi Salim and Ashok Pandey*

Biotechnology Division, National Institute for Interdisciplinary Science and Technology (NIIST), CSIR, 695 019 Trivandrum, Kerala, India

Article history:
Received November 20, 2010
Accepted January 25, 2011

Key words:
A. niger phytase, inducer concentration, recombinant appA

Summary:
Microbial phytases, especially from fungal and bacterial sources, have received much attention as food additives in human nutrition and as feed supplements for monogastric animals. An effective expression screening method for recombinant production of this enzyme on a small scale is industrially desirable. An effort has been made in this work to clone and express phytase genes from Aspergillus sp. and Escherichia coli with the selected host, vector and inducer combination. Albeit the formation of insoluble inclusion bodies by fungal phytase, recombinant E. coli appA was effectively expressed in a cost-effective manner in the periplasm of BL21plysS using an inducer concentration of 0.01 mM in 4 h of growth. Enzyme was purified in three consecutive steps and functional studies were carried out.

*Corresponding author:    This email address is being protected from spambots. You need JavaScript enabled to view it., This email address is being protected from spambots. You need JavaScript enabled to view it.
                                         ++91 471 251 5279
                                         ++91 471 249 1712