Abstract
Enhanced production of recombinant dengue multi-epitope protein in Escherichia coli was achieved by optimization of culture medium. Complex media (Luria Bertani broth, Terrific broth, Super broth, M9 minimal media, five times Luria Bertani broth, semi-defined medium enriched with tryptone and yeast extract, and semi-defined medium enriched with glucose) were evaluated for production of recombinant dengue multi-epitope protein in shake flasks. The recombinant protein was further produced by fed-batch fermentation using 5 L bioreactor. Cells were grown in optimized semi-defined medium, and feeding was carried out with 5X medium and glycerol. When growth reached 14.35 g/L of dry cell weight, culture was induced with 0.5 mM IPTG and further grown for 4 h to reach 18.37 g/L dry cell weight. The recombinant dengue multi-epitope protein was purified from inclusion bodies under denaturing conditions using metal affinity chromatography, which yielded 96.43 mg/L of protein. The purified protein was found to be reactive with dengue-infected human serum samples.
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