Enhanced Production, Purification, and Characterisation of Recombinant Taq DNA Polymerase

Infectious diseases have become one of the major health issues in the 21st century. Taq DNA polymerase is a critical component of polymerase chain reaction. To meet the requirement of this enzyme in any biological emergency like Covid-19, recombinant Taq DNA polymerase was developed and scale-up produced using Escherichia coli. In this study, gene encoding recombinant Taq DNA polymerase was cloned, and expressed using E. coli. Optimisation of process parameters was done to achieve high expression of the enzyme. Further, bioreactor production of this enzyme was also carried out using optimised parameters to attain large amount of biomass for subsequent purification. A simple purification strategy using affinity chromatography followed by diafiltration was used to get this enzyme in pure form. The purified Taq DNA polymerase was characterised by SDS-PAGE and Western blot analysis. Further, this enzyme was used in PCR for confirmatory diagnosis, and the result of PCR assay confirmed its usefulness for the diagnosis of disease. The present study established that the recombinant Taq DNA polymerase produced indigenously may be potentially deployed for disease diagnosis in PCR during any outbreak scenario as well as laboratory settings.