| Summary: | The present study aimed to produce a biosurfactant from <i>Candida</i> yeast cultivated in a low-cost medium made of sugar-cane molasses (5%), frying oil waste (5%), and corn steep liquor (5%). Initially, the production at the flask-scale was investigated and then scaled up in bioreactors to 1.2, 3.0, and 50 L to simulate a real production scale. The products obtained an excellent reduction in surface tensions from 70 to 29 mN·m<sup>−1</sup> in the flask-scale, comparable to 33 mN·m<sup>−1</sup> in the 1.2-L reactor, to 31 mN·m<sup>−1</sup> in the 3-L reactor, and to 30 mN·m<sup>−1</sup> in the 50-L reactor. Regarding the yield, it was observed that the isolation by liquid-to-liquid extraction aided biosurfactant production up to 221.9 g·L<sup>−1</sup> with a critical micellar concentration of 0.5%. The isolated biosurfactant did not exhibit an inhibitory effect on the germination of vegetable seeds and presented no significant acute toxicity in assays with <i>Artemia salina</i> and <i>Allium cepa</i>. Among the different formulations of mayonnaise-like sauces, the most stable formula was observed with the addition of the biosurfactant at a concentration of 0.5% and the greatest results were associated with the guar and carboxymethyl cellulose gums. Thus, the biosurfactant from <i>C. bombicola</i> represents a promising alternative as a food additive in emulsions.
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