In this study, sugarcane bagasse hemicellulosic hydrolisate was evaluated as carbon source in biosurfactant production by yeasts. 27 yeasts were screened in culture media based on commercial xylose, non-detoxified and detoxified sugarcane bagasse hydrolysate to select potential strains for BS production with emulsifying and tensoactive properties. Cutaneotrichosporon mucoides UFMG-CM-Y6148 cultured in medium based on detoxified hydrolysate was the best producer of biosurfactant with a volumetric productivity of 0.167 g.L−1. h−1 and highlighted emulsifying properties in kerose as emulsification index higher than 50% and an emulsifying activity of 1.0 EU. mL−1, and its effectiveness was demonstrated over a wide range of temperature, pH and salt concentration. The chromatography and spectrometry analysis demonstrated that the BS produced by C. mucoides UFMG-CM-Y6148 was a sophorolipid. The results obtained in this study demonstrated the successful sustainable prodution of biosurfactant by yeasts in media with hemicellulosic hydrolysate and the viability of these products for biorefineries.