Production of recombinant nonstructural 1 protein in Escherichia coli for early detection of Japanese encephalitis virus infection
Article first published online: 27 MAR 2012
© 2012 The Authors. Microbial Biotechnology © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd
Thematic Issue: Bioterrorism Research
Volume 5, Issue 5, pages 599–606, September 2012
How to Cite
Tripathi, N. K., Kumar, J. S., Biswal, K. C. and Rao, P. V. L. (2012), Production of recombinant nonstructural 1 protein in Escherichia coli for early detection of Japanese encephalitis virus infection. Microbial Biotechnology, 5: 599–606. doi: 10.1111/j.1751-7915.2012.00344.x
- Issue published online: 23 AUG 2012
- Article first published online: 27 MAR 2012
- Received 20 January, 2012; revised 24 February, 2012; accepted 27 February, 2012.
Japanese encephalitis is a major public health problem in South-East Asia and Western Pacific countries. The recombinant nonstructural 1 (rNS1) protein of Japanese encephalitis virus is a potential diagnostic as well as vaccine candidate. Developments of cost-effective and simple culture media as well as appropriate culture conditions are generally favourable for large-scale production of recombinant proteins. The effects of medium composition and cultivation conditions on the production of rNS1 protein were investigated in shake flask culture as well as batch cultivation of Escherichia coli. Further, the fed-batch process was also carried out for high cell density cultivation (HCDC) of E. coli expressing rNS1 protein. Isopropyl-β-d-thiogalactopyranoside (IPTG) was used to induce the expression of rNS1 protein at ∼ 13 g dry cell weight per litre of culture. The final dry cell weight after fed-batch cultivation was ∼ 17 g l−1. The Inclusion bodies were isolated and purified through affinity chromatography to give a final product yield of ∼ 142 mg l−1. The reactivity of purified protein was confirmed by Western blotting and Enzyme linked immunosorbent assay. These results show that rNS1 protein may be used as a diagnostic reagent or for further prophylactic studies. This approach of producing rNS1 protein in E. coli with high yield may also offer promising method for production of other viral recombinant proteins.