Team:REC-CHENNAI/Characterization-Description
CHARACTERIZATION DESCRIPTION
The 2017 team of iGEM REC-CHENNAI, had worked on food preservation by exploiting the antimicrobial property of Latarcin-2a_F10K in their model. However, the team had not provided validated results for the mutant’s decreased haemolytic and cytotoxic activities while retaining the antimicrobial activity.
We characterised Latarcin-2a_F10K to confirm whether the mutant had retained the antimicrobial action of the wild type while displaying decreased haemolytic and cytotoxic actions. For this, Latarcin-2a_F10K was synthesized, purified and quantified. The retention of the antimicrobial activity in the mutant as in the wild type against E. coli, B. subtilis, and S. cerevisiae was confirmed by observing similar Minimum Inhibitory Concentration (MIC), while the reduced haemolytic activity of the mutant Latarcin vis-a-vis the wild type was confirmed by their action on bovine blood at various concentrations. It was observed that mutant Latarcin showed no haemolytic action for a scale of concentrations as that of native Latarcin. Further, MTT assay performed on mammalian cell lines (HeLa and C212 skeletal muscle cell lines) treated with mutant and native Latarcin demonstrated decreased cytotoxicity of the mutant with respect to the native Latarcin.
