Conventional biosurfactants have high production costs. Therefore, the use of low-cost carbon sources for their production is attractive for industry. The ability to remain stable under various environmental conditions further extends industrial application. Here we aimed to evaluate the stability of a new lipopeptide produced by Corynebacterium aquaticum using fish residue as an unconventional energy source. The biosurfactant was produced using 3% fish residue, 2% of the microorganism, and mineral medium. Biosurfactant characterization was performed by thin layer chromatography (TLC), as well as by testing its infrared, surface tension, emulsifying activity, and ionic character. The stability of the biosurfactant was evaluated by testing its surface tension at a range of temperatures, pH, and saline concentrations, as well as after 6 months of storage. The biosurfactant was characterized as a lipopeptide due to its retention time, which was coincident with the amino acid and lipid chains obtained in the TLC analysis, being confirmed by some regions of absorption verified in the infrared analysis. The surface tension and emulsifying activity of the biosurfactant were 27.8 mN/m and 87.6%, respectively, and showed anionic character. The biosurfactant was stable at temperatures of 20 to 121 °C, in saline concentrations of 1 to 7%, and at pH close to neutrality. Based on our findings, it is possible to use unconventional sources of energy to produce a lipopeptide biosurfactant that can act under various environments.