Biosurfactants are amphiphilic molecules produced by microorganisms, particularly bacteria. Biosurfactants have many applications including the bioremediation of hydrocarbon pollution in the environment. Hence, the toxicity of biosurfactants produced from bacteria should be evaluated. In this report, a biosurfactant harvested from the Pseudomonas sp. was characterized in terms of its ability to form an emulsion layer with hydrocarbons, its biochemical content and its toxicity towards germinating seeds. The biosurfactant produced was characterized as a glycolipid since a positive result was obtained in the sugar assay and a negative result was obtained in the protein assay. The ability of the biosurfactant to emulsify engine oil was found to be as strong as that of Triton X. Seeds of mung beans were germinated using the biosurfactant to assess the phytotoxicity of the biosurfactant. The biosurfactant was found not to have toxic effects on the development of roots and shoots of mung bean. The calculated germination index (GI) value of the mung bean exposed to the biosurfactant was significantly higher than that of the mung bean exposed to the chemical surfactant Triton. Hence, it can be concluded that the biosurfactant is non-toxic and exhibits non-inhibitory effects on the growth of mung bean and fenugreek and can be applied in the environmental bioremediation process.