Canola flake was investigated as a potential substrate for fungal conversion to produce polyunsaturated fatty acids (PUFAs)-added oil in a 7 1 fermenter. The results showed that yields of total oil were reduced 9-22% compared to initial oil in the canola flake, but as high as 445 mg/l arachidonic acid (ARA, C20:4n6) and 67 mg/l eicosapentaenoic acid (EPA, C20:5n3) were produced. The percentages of ARA and EPA of total fatty acids in this fermented oil were 15.5% and 2.3%, respectively. Supercritical CO2 extraction was then investigated for the lipid recovery from fermented canola flake, and extraction kinetics were modeled. The feasibility was demonstrated for production of PUFAs in a laboratory-scale fermentor using canola flake as a single nutrient, and for lipid extraction using supercritical CO2. (c) 2008 Elsevier Ltd. All rights reserved.