A bottleneck in the production of biodiesel from microalgae is the dewatering and lipid extraction process which is the dominant energy penalty and cost. A novel biodiesel production platform based on vortex fluidic device (VFD)-assisted direct transesterification (DT) of wet microalgal biomass of Chloroparva pannonica was developed and evaluated. Fatty acid extraction and fatty acid to FAME conversion efficiencies were used at different parameter settings to evaluate performance of the processing technology in confined and continuous mode. A response surface method based on Box-Behnken experimental design was used to determine the effects of water content, the ratio of biomass to methanol and residence time in the VFD. Average extraction efficiencies were 41% and conversion efficiencies >90% with the processing technology showing a broad tolerance to parameter settings. The findings suggest that VFD-assisted DT is a simple and effective way to produce biodiesel directly from wet microalgae biomass at room temperature.