A semi-pilot continuous process (SPCP) for enzymatic biodiesel synthesis utilizing near-critical carbon dioxide (NcCO₂) as the reaction medium was developed with the aim of reducing the reaction time and alleviating the catalyst inhibition by methanol. Biodiesel synthesis was evaluated in both lab-scale and semi-pilot scale reactors (batch and continuous reactors). In a SPCP, the highest conversion (∼99.9 %) in four and a half hours was observed when three-step substrate (methanol) addition (molar ratio [oil/methanol] = 1:1.3) was used and the reaction mixture containing enzyme (Lipozyme TL IM, 20 wt.% of oil) was continuously mixed (agitation speed = 300 rpm) at 30 °C and 100 bar in a CO₂ environment. The biodiesel produced from canola oil conformed to the fuel standard (EU) even without additional downstream processing, other than glycerol separation and drying.