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As the cosmetic market continues to transform, innovative ingredients such as biosurfactants (BS) emerge as a fundamental step towards a generation of cosmetic products characterized by efficacy, safety, and environmental responsibility. In alignment with the global push towards natural, renewable, and non‐toxic materials, BS stands out as an interesting alternative to conventional synthetic surfactants. BS are a type of green surfactants and compared to synthetic surfactants derived from petroleum they are more biodegradable and have low or no toxicity. This work aimed to investigate the suitability of a biosurfactant extracted from the yeast Scheffersomyces shehatae (BS‐SS) to be used in cosmetic applications. The cytotoxicity of the BS‐SS was tested in fibroblast (L929) and keratinocyte (HaCaT) cell lines and the potential for ocular irritation was assessed by the hen's egg chorioallantoic membrane test (HET‐CAM). The cleaning efficiency of the BS‐SS was measured as the ability of make‐up removal in pig skin. Our results showed that the IC 50 values were 10.49 mg/mL for fibroblasts and 11.77 mg/mL for keratinocytes, and the biosurfactant did not result in hemorrhage, lysis, and coagulation damages in the chorioallantoic membrane, which classifies the biosurfactant solution as non‐irritant. Therefore, ocular toxicity and in vitro skin cell toxicity results showed that BS‐SS can be tolerable and safe for cosmetic purposes. BS‐SS featured promising cleaning properties, similar to the cleansing activity of the 1% SDS solution and of micellar water for lipstick and foundation products. In conclusion, the results indicate its potential use in cosmetic applications. [ABSTRACT FROM AUTHOR]