Biosurfactants are nontoxic, renewable, and environmentally benign; and have excellent emulsification and antibacterial properties. Oleic acid‐rich oils have a crucial role as a hydrophobic substrate in the biological production of sophorolipids (SL). The major purpose of this study is to identify a low‐cost oleic acid‐rich hydrophobic substrate that may be used as a secondary carbon source in the synthesis of SL by Starmerella bombicola (S. bombicola). Thus, the biosynthesis of SL was investigated utilizing a variety of edible and nonedible oils rich in oleic acid, which acts as a precursor. Characterization of SL generated from different oils using FT‐IR and NMR indicated the presence of both lactonic and acidic SL. SL production was greatest with secondary carbon sources, with 38.9, 27.57, and 19 g/L, respectively, in the order olive oil > sunflower oil > fleshing oil. Surprisingly, the cost analysis for fleshing oil synthesis indicated that it was the lowest at 0.43 INR/g, followed by sunflower oil at 0.52 INR/g and olive oil at 1.71 INR/g. Our findings indicate that fleshing oil is a potential secondary carbon source with significant SL yield as well as environmental and economic benefits.

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