Fungal enzymes are widely used in technological processes and have some interesting features to be applied in a variety of biosynthetic courses. Here, free and substrate-immobilised lipases fromFusarium verticillioidesP24 were obtained by solid-state fermentation using wheat bran as substrate and fungal carrier. Based on their hydrolytic and transesterification activities, the lipases were characterised as pH-dependent in both reactions, with higher substrate conversion in an alkaline environment. Thermally, the lipases performed well from 30 to 45 degrees C, being more stable in mild conditions. Organic solvents significantly influenced the lipase selectivity using different vegetable oils as fatty acid source. Omega(omega)-3 production inn-hexane achieved 45% using canola oil, against approximate to 18% in cyclohexane. However, omega-6 production was preferably produced for both solvents using linseed oil with significant alterations in the yield (approximate to 79% and 49% forn-hexane and cyclohexane, respectively). Moreover, the greatest enzyme selectivity for omega-6 led us to suppose a lipase preference for the Sn1 position of the triacylglycerol. Lastly, a transesterification reaction was performed, achieving 90% of ester conversion in 72 h. This study reports the characterisation and use of free and substrate-immobilised lipases fromFusarium verticillioidesP24 as an economic and efficient method for the first time.