Fuel and oil spills during the exploration, refining, and distribution of oil and petrochemicals are primarily responsible for the accumulation of organic pollutants in the environment. The reduction in contamination caused by hydrocarbons, heavy metals, oily effluents, and particulate matter generated by industrial activities and the efficient recovery of oil at great depths in an environmentally friendly way pose a challenge, as recovery and cleaning processes require the direct application of surface-active agents, detergents, degreasers, or solvents, often generating other environmental problems due to the toxicity and accumulation of these substances. Thus, the application of natural surface-active agents is an attractive solution. Due to their amphipathic structures, microbial surfactants solubilize oil through the formation of small aggregates (micelles) that disperse in water, with numerous applications in the petroleum industry. Biosurfactants have proven their usefulness in solubilizing oil trapped in rock, which is a prerequisite for enhanced oil recovery (EOR). Biosurfactants are also important biotechnological agents in anti-corrosion processes, preventing incrustations and the formation of biofilms on metallic surfaces, and are used in formulations of emulsifiers/demulsifiers, facilitate the transport of heavy oil through pipelines, and have other innovative applications in the oil industry. The use of natural surfactants can reduce the generation of pollutants from the use of synthetic detergents or chemical solvents without sacrificing economic gains for the oil industry. Therefore, investments in biotechnological processes are essential. It is predicted that, in the not-too-distant future, natural surfactants will become viable from an economic standpoint and dominate the world market. The application of biosurfactants in these settings would lead to industrial growth and environmental sustainability. The main goal of this paper is to provide an overview of diverse applications of biosurfactants on environmental remediation, petroleum biotechnology, and the oil industry through a scientific literature review.