Marine biofilms are a virtually untapped source of bioactive molecules that may find application as novel antifoulants in the marine paint industry. This study aimed at determining the potential of marine biofilm bacteria to produce novel biomolecules with potential application as natural antifoulants. Nine representative strains were isolated from a range of surfaces and were grown in YEB medium and harvested during the late exponential growth phase. Bacterial biomass and spent culture medium were extracted with ethanol and ethyl acetate, respectively. Extracts were assayed for their antifouling activity using two tests: (1) antimicrobial well diffusion test against a common fouling bacterium, Halomonas marina, and (2) anti-crustacean activity test using Artemia salina. Our results showed that none of the ethanolic extracts (bacterial biomass) were active in either test. In contrast, most of the organic extracts had antimicrobial activity (88%) and were toxic towards A. salina (67%). Sequencing of full 16 S ribosomal DNA analysis showed that the isolates were related to Bacillus mojavensis and Bacillus firmus. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) profiling of ethyl acetate extracts of culture supernatants showed that these species produce the bioactive lipopeptides surfactin A, mycosubtilin and bacillomycin D.