The cosmetic industry provides a wide variety of shampoos to treat dandruff, containing insoluble ingredients such as Zn pyrithione. However, the solubility of this active ingredient is quite limited in both water and oil media; thus, antidandruff formulations must include a large amount of chemically synthesized stabilizing ingredients to avoid their precipitation. In this work, the stabilization of Zn pyrithione in O/W emulsions using a biosurfactant (BS) extract and Tween 80 is studied. The study includes an incomplete factorial design based on the tea tree oil/water ratio and both surfactant and biosurfactant concentrations. The formulations are characterized in terms of particle size, stability after 30 days, and solubility of Zn pyrithione. The formulation that provided the most favorable results contains Tween 80 (5%) and BS extract (2.5%), with an O/W ratio of 0.01. This provides the smallest particle size (40.5 μm), good stability after 30 days (91.0%), and the highest solubility of Zn pyrithione (59%). The results obtained enable the use of the combination of BS extract and Tween 80 as bio/surfactants of antidandruff shampoo formulations, along with another antiseptic agent such as tea tree oil. Furthermore, this is the first work where a biosurfactant is considered to be a stabilizing agent in antidandruff formulations.