Biosurfactants are surface-active compounds comprised of both hydrophilic and hydrophobic moieties. These biomolecules are produced by fungi, yeast, and bacteria. Fungal biosurfactants have gained considerable attention for various applications due to their unique structures and properties. Fungi are a promising candidate for producing these biomolecules, such as sophorolipids, xylolipids, hydrophobins, polyol lipids, cellobiose lipids, and mannosylerythritol lipids. In this chapter, we have focused on three critical aspects. First, we present the issues related to various types of fungal biosurfactants, physicochemical properties, and their application. Next, we have discussed the culture condition, cultivation parameters, and process optimization. In addition, parameters involved in the sustainable production of biosurfactants, such as simultaneous production and downstream processing, were considered. Finally, advanced achievements in biosurfactant applications, in addition to some aspects of commercial fungal-based biosurfactant formulations, have been presented over the years.