The enzymatic synthesis of biodiesel by a high-pressure semi-continuous process in near-critical carbon dioxide (NcCO(2)) was studied. Biodiesel synthesis was evaluated in both batch and semi-continuous systems to develop an effective process. Batch processing demonstrated the advantageous properties of NcCO(2) as an alternative reaction medium. Three immobilized lipases (Novozym 435, Lipozyme RM IM, and Lipozyme TL IM from Novozymes) were tested, with Lipozyme TL IM the most effective, showing the highest conversion. Biodiesel conversion from several edible and non-edible oil feedstocks reached >92%. Higher conversion (99.0%) was obtained in a shorter time by employing repeated batch processes with optimized conditions: 44.3 g (500 mM) canola oil, a substrate molar ratio (methanol:oil) of 3:1, an enzyme loading of 20 wt% (of the oil used), at 30 °C, 100 bar, and 300 rpm agitation. The enzyme maintained 80.2% of its initial stability after being reused eight times. These results suggest that this method produces biodiesel energy-efficiently and environment-friendly.
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