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Biosurfactants are amphipathic molecules, biodegradable, with reduced toxicity. They can be synthesized by fermentative processes from oleaginous compounds and agro-industrial by-products. In this context, the present study describes the production and the physical, chemical, and structural characterization of the bioemulsifier secreted by the yeast <i>Scheffersomyces shehatae</i> 16-BR6-2AI in a medium containing hemicellulosic sugarcane bagasse hydrolysate combined with soybean oil. The bioemulsifier was produced in Erlenmeyer flasks and isolated; then, the physicochemical and structural characterization of the formed molecule was carried out. The following fermentation parameters were obtained: Y_X/S = 0.45, Y_P/S = 0.083, and productivity of 0.076 g/L/h. The bioemulsifier was found to be a polymer containing 53% of carbohydrates, 40.92% of proteins, and 6.08% of lipids, respectively. The FTIR spectrum confirmed the presence of functional groups such as amides, amines, and carbonyls. The bioemulsifier was stable over a range of temperature (−20 °C to 120 °C), salinity (1–15%), and pH (2–12). It was observed that the biomolecule has a better emulsifying action in organic solvents with a non-polar character. Therefore, this biomolecule is a potential substitute for synthetic surfactants and can be used in different applications.