tThis work aimed to study the production of a biosurfactant by a new strain of Bacillus subtilis ICA56 isolated from a Brazilian mangrove and to evaluate its functional properties and applicability for biore-mediation. The use of agro-industrial wastes (glycerol, sunflower oil, cheese whey and cashew applejuice) as alternative substrates for biosurfactant production was tested as this may lead to a reductionin the cost of the bioprocess. Glycerol was the best carbon source yielding 1290 mg L−1of crude biosur-factant. The critical micellar concentration of the crude biosurfactant produced by ICA56 was 25 mg L−1and, at this concentration, it was able to reduce the surface tension of the water from 72 to 30 mN m−1and to reduce the interfacial tension on a water/gasoline system from 15 to 3 mN m−1. Furthermore, thecrude biosurfactant retained its tensoative properties in a broad range of pH, temperature and salinity and it was not toxic to Artemia salina. In this work, model experiments were conducted to simulate theremoval of hydrocarbons and heavy metals from contaminated environmental systems in the laboratoryby the crude biosurfactant produced by ICA56. Results showed that it was very efficient, highlighting itspotential for bioremediation