Biodiesel, a promising type of biofuel, can be produced from various types of renewable feedstocks, ranging from animal fats to plant oil. It is mainly made up of fatty acid alkyl ester compounds due to the transesterification reaction. This work aims to synthesize and characterize biodiesel, known as fatty acid methyl esters, from canola oil using an enzymatic reaction involving immobilized Novozym 435 and Rhizomucor miehei (RM IM) lipase enzymes.4 g of canola oil was added to the reaction mixture consisting of 0.2 g immobilized lipase and 3:1 methanol to oil ratio. First, the enzymatic methanolysis reaction was conducted at the temperature of 35°C and at agitation rate of 216 rpm for 24 hours. Next, the synthesized biodiesel was characterized using the Gas Chromatography-Mass Spectroscopy (GC/MS) analysis. Based on the analysis results, the main fatty acid methyl esters present in both products were hexadecanoic acid, 9-octadecenoic acid (z)-, 9,12,15-octadecatrienoic acid, (z, z, z)-, and 11,14-eicosadienoic acid. The transesterification of canola oil using both enzymes consistently revealed methyl oleate as the methyl ester with the highest composition, ranging from 67 to 71 %. In conclusion, canola oil was successfully converted into fatty acid methyl ester via the enzymatic transesterification process in this study.