Enduracidin is a 17-amino acid lipopeptide antibiotic produced by Streptomyces fungicidicus. The peptide acts by binding to extracellular Lipid II and exhibits potent bactericidal activity against a wide variety of Gram-positive bacteria, including vancomycin resistant strains. In studying the biosynthesis of enduracidin we have identified the enduracidin gene cluster and have begun investigating the formation of the arginine-derived amino acid enduracididine, the timing of chlorination events and the steps leading to the 2Z,4E branched chain lipid tail. Results from targeted gene disruptions, chemical and genetic complementation studies, and findings from labeled precursor incorporation experiments will be presented.