Fluorescent labeling of nisin Z and assessment of anti-listerial action

Abstract

Biomolecule labeling by fluorescent markers has emerged as an innovative methodology for bio-analytical purposes in foodmicrobiology, medicine and pharmaceutics due to the great advantages of thismethod such as precision, wide detection limits, and in vivo recognition. Fluorescent nisin Z was synthesized by linking the carboxyl group and amino group of nisin Z and 5-aminoacetamido fluorescein (AAA-flu). This new structure was fully characterized by mass spectrometry with a molecular weight of 3717.3 Da. Intracellular K+ leakage and transmembrane electrical potential (Δψ)were used to evaluate the antibacterial action of the labeledmolecule against three listerial strains and demonstrated that nisin Z endured the labeling processwithout any activity loss. In vivo activity of labeled nisin was observed by confocal laser microscope which revealed its localization at the septum of listerial cell division sitewhere themembrane-bound cellwall precursor lipid II ismaximal. Fluorescent nisin Z showed its great potential as a tool to study antibacterial mechanism of action of nisin in biological systems