The search for renewable and sustainable fuels is becoming increasingly more important. As a result, the use of biomass as an energy source is one of the most promising methods in the reduction of energy dependence on non-renewable fossil fuels. In order to reduce the amount of solvent used during direct transesterification, this study has aimed at producing biodiesel from microalgae (Chlorella vulgaris) biomass using enzymatic catalysis (Novozym 435) and pressurized fluid (propane). The effects of pressure (3000 to 8000 kPa), temperature (323.15 to 353.15K), and exposure time (1 to 8h) have been studied through the experimental design for esters conversion. The newly investigated system showed consistency through the experimental design technique, evaluating the effect of significant variables such as time, temperature, and pressure in the process. The good conversions yielded have been achieved using mild temperature and higher pressure conditions, demonstrating that this process has great potential for obtaining biodiesel from microalgae biomass.