Nisin is an antimicrobial peptide with bacterial, fungicidal, virucidal properties, attacking bacteria and destroying the cell membranes. Thanks to its stability to hard conditions, it is a candidate for the use as molecular recognition elements in biosensing platform. In this work, the use of nisin as a biological molecule for the development of a sensitive biosensor for bacteria detection is reported: nisin molecules were immobilised on gold electrodes and Electrochemical Impedance Spectroscopy was to investigate the electrochemical responses after the exposure of the biosensor to different bacteria. The biosensor was able to detect all bacterium tested with different impedimetric responses; the singular impedimetric behaviours recorded after the exposure to pathogenic and non - pathogenic Salmonella strains, highlighted the possibility of the proposed biosensor to detect selectively Salmonella cells with a low limit of detection of 1.5 * 10 CFU/mL. Finally, the developed biosensor was used to detect Salmonella in milk.