Characterization of Recombinant Thermostable Phytase from Thermotoga naphthophila: A Step for the Fulfilment of Domestic Requirement of Phytase in Pakistan
Characterization of Recombinant Thermostable Phytase from Thermotoga naphthophila: A Step for the Fulfilment of Domestic Requirement of Phytase in Pakistan
Furqan Sabir1, Muhammad Tayyab1,*, Bushra Muneer2, Abu Saeed Hashmi1, Ali Raza Awan1, Naeem Rashid3, Muhammad Wasim1 and Sehrish Firyal1
ABSTRACT
Supplementation of feed with phytase is the most suitable strategy for the availability of free phosphorus for the growth of monogastric poultry birds. Current study deals with the production and characterization of recombinant thermostable phytase from Thermotoga naphthophila (PHYTN). This study was an initial step for the fulfilment of domestic industrial requirement of phytase in Pakistan.The PCR resulted in the amplification of 1.8 kb phytase gene. SDS-PAGE confirmed the size of recombinant protein as 70 kDa. The optimization studies demonstrated the maximal production of recombinant phytase, when the recombinant cells were induced with 1.4 mM IPTG with the post induction time of 6 hours. PHYTN showed maximal activity at 80°C in 50 mM sodium acetate buffer pH 6. Presence of Fe3+ or Cu2+ showed an enhancing effect on the PHYTN activity. Thermostability studies demonstrated that PHYTN retains 88% residual activity when the protein was incubated at 80°C for 1.5 h in the presence of 1.5 mM Fe3+. The enzyme exhibited Km and Vmax values of 50 mM and 2500 µmole/min respectively when sodium phytate was used as substrate. The stability of enzyme at a wide range of temperature and pH makes it a potential candidate to be used in the poultry feed industry.
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December 2017
Vol. 49, Iss. 6, Pages 1937-2341
