Surfactin is a mixture of isoforms and homologous seriespresenting some differences in their physicochemical propertiesmainly caused by variations in the chain length (C12–C16) and branching of its hydroxy fatty acid component as well as replace-ments of the amino acid components on the peptide ring [9].These variations might be genetically determined, depend on thestrain, or even on the operational conditions used for biosurfac-tant production [9]. The differences on physicochemical propertieson homologues series and isoforms of biosurfactants might affect the application and effectiveness of these biomolecules, then beingnecessary to evaluate its functional properties, toxicity and suit-ability for industrial application [10].In this context, this study aimed to evaluate the production of biosurfactant by a new strain of Bacillus subtil is ICA56, grown on low cost substrates, analyze its functional properties under different abiotic variables, and test its potential to remove hydrocarbons and metals.