Marine bacteria are a rich source of potentially useful antimicrobial molecules. However, much of the microbial diversity in marine ecosystems with its potential for uncovering new antimicrobial compounds remains to be discovered. This is particularly true for surface-attached marine bacteria, which comprise microbial communities that are generally unique to a host surface and geographic location. The current study characterises culturable microbial communities on marine surfaces from Sydney Harbour, Australia, and tests their antimicrobial activities. A high proportion (47%) of the 104 marine isolates from Sydney Harbour could not be classified to a known genus based on 16S ribosomal RNA gene sequences. Assays of antimicrobial activity from the 104 isolates showed that antimicrobial production is not widespread throughout the phylogeny of isolates with 8 of the 10 antimicrobial producers clustering into a distinct phylogenetic clade. These 8 closely related antibacterial isolates had potent activity in antibacterial cross-dilution assays, with no growth of target bacteria at supernatant concentrations of less than 6.6 % v/v. To gain an insight into the types of molecules responsible for this potent activity, differential polarity extractions were carried out on antibacterial culture supernatants from these 8 isolates. All of the activity fractionated into the most polar phase, suggesting that the antibacterial molecules are highly polar. Proteolytic digestion inhibited activity, indicating that the antibacterial molecules were proteins. This study is the first to link the phylogeny of numerous surface-attached marine bacteria with antimicrobial production.