A novel Amycolatopsis strain KCTC 29142 was isolated and characterized based on the polyphasic taxonomic analysis including morphological observation, phylogenetic analysis, physiological and chemotaxonomic characteristics. The ethyl acetate extract of strain KCTC 29142 culture broth showed strong antibacterial activity and the active compound was identified as siderochelin A, a ferrous-ion chelating compound. In this study, siderochelin A showed good activity against multi-drug resistant pathogens, including Acinetobacter baumanii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), and Escherichia coli (E. coli ). The minimum inhibitory activity against clinical isolates was also determined.