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This present work aims to overcome some problems that follow the production of biodiesel by chemical methods. In this framework, we have substituted all the reagents used in this method in order to produce a biodegradable, renewable biofuel with less impurities and side products. Thus we substituted fresh vegetable oil by used frying oil, methanol by dimethyl carbonate (DMC) which is a non-toxic and biodegradable reagent, chemical catalysis by biocatalysis (enzymatic catalysis). We did not also use an organic solvent since DMC plays the role of acyl acceptor and solvent.The results found in this work show that biodiesel can be synthesized in good yield and without impurities from used frying oil using lipase as catalyst and DMC as acyl acceptor and solvent at the same time at a temperature not exceeding 45°C.