In this study, the optimal process conditions for the production of biodiesel from Gmelina arborea seed oil were investigated using the transesterification method. The impacts of process parameters such as catalyst concentration, reaction time, reaction temperature, methanol/sample ratio, and agitation speed were investigated separately and, in a matrix, to determine synergistic relationships. The response surface methodology was used to investigate the biodiesel yield and determine if it would be suitable for replacing fossil fuels. A two-level, five-factorial approach was used to investigate the impacts of process parameters on biodiesel yield. The results showed that the greatest biodiesel production of 78.28% was achieved at a catalyst concentration of 3%(W), a methanol/oil ratio of 10:1, a reaction temperature of 60OC, a reaction time of 3 hours, and an agitation speed of 300 rpm.