Vitamin E deficiency has caused many problems for humans, especially patients who have difficulty absorbing this substance in their meals. Vitamin E supplements are commonly suggested to patients to solve these problems. Thus, improving the bioavailability of vitamin E in O/W microemulsions is one of the challenges for researchers. Another challenge is the safe production of these materials which must be considered. Using biosurfactants instead of surfactants is a move towards green and safe production and consumption of these materials. This paper discusses method of preparing vitamin E microemulsions by using a crude biosurfactant from Yarrowia lipolytica IMUFRJ50628. In this work, maximal crude biosurfactant concentration 3.3 g/L was attained after 130 h of cultivation and the crude biosurfactant was characterized by FTIR analysis. Phase diagram of bio‐ based microemulstions (water/vitamin E/soy bean oil/lecithin/biosurfactant) was compared with chemical‐based microemulstions (water/vitamin E/soy bean oil/lecithin/Tween 80). The results showed that the crude biosurfactant is a good alternative to the Tween 80 for green production consideration. Dynamic light scattering measurements for the both systems showed they were stable under human stomach conditions such as 37°C and pH = 2 for 7 days. Our successful method for preparing a bio‐ based O/W microemulsion confirmed that biosurfactants can be used instead of surfactants from chemical synthesis and it is thought that the incorporation of biosurfactants may be widely used in the future as a vehicle for vitamin E in the pharmaceutical and food industries.