Expression and purification of modified recombinant Nisin in Escherichia coli BL21

Introduction and Objective: Nisin is an antibacterial peptide produced by the bacterium lactococcus lactis. A series of recent studies has shown that natural nisin has selective cytotoxicity towards cancer cells and could be a potential anti-cancer agent. In this study we aimed to produce a modified recombinant Nisin, containing two antibacterial peptides, which can be used in future studies investigating its anti-cancer effects.Materials and Methods: At first, a recombinant gene cassette containing Nisin A encompassed by two bacterial anti-cancer peptide sequences was ordered to Biomatik Corp. The cassette was transformed to E coli BL21 and, using IPTG, the modified Nisin gene expression was induced. We then examined the resultant protein by the SDS PAGE gel analysis, followed by confirmation with western blotting. Finally, the intended protein was purified by Ni-resin chromatography column and then confirmed by western blot analysis.Results: After induction with IPTG, an expected band of 12.5 kDa corresponding to the desired protein was observed on the SDS PAGE gel. Also, after purification, the production of modified protein was confirmed by the presence of a single band on the SDS PAGE gel followed by western blot analysis.Conclusion: In this study, the modified recombinant Nisin was expressed in E.coli BL21 and then successfully purified, which can be used in studies aimed to investigate the this fusion peptides‟ anti-cancer activity.

کلیدواژه ها:

lactococcus lactis ، Nisin A ، antibacterial peptide ، anti-cancer ، recombinant peptide