The present study assesses the Paenibacillus sp. D9 lipopeptide biosurfactant synthesis in cheap substrates including functional properties and applicability for varying biotechnological processes. Different experimental setups were made for oil dispersion, heavy metals removals from contaminated environments, and washing performance. The study revealed surface tension activities of 31.7–32.7 mN/m, and maximum biosurfactant yield of more than 8 g/L. Removals of 85.90%, 98.68%, 99.97%, 63.28%, 99.93%, and 94.22% were obtained for Ca, Cu, Fe, Mg, Ni, and Zn, respectively from acid mine effluents. In comparison with chemical surfactants, there was pronounced removal of heavy metals from wastewater, contaminated sands, and vegetable matter, as well as improved oil dispersing activity. A comparative study revealed that biosurfactant was more efficient (> 60%) for removal of tomato sauce and coffee stains than chemical surfactants (< 50%). Thus, lipopeptide biosurfactants are green biomolecules reducing hazards and contaminations within the environment. The future use of this lipopeptide biosurfactant is greatly promising in biotechnology.