Competing between foods and biofuels production, the world food crisis and high oil prices have ignite attention in algaculture for making biofuels. Among algae biodiesel positive characteristics are: it can be produced using sea water and wastewater without affecting fresh water resources, is biodegradable and relatively harmless to the environment if spilled. This work represents the first attempt of optimizing thirdgeneration biodiesel production by homogeneous transesterification. It was used a synthetic algae oil based on Chorella vulgaris fatty acid composition. The first step of this research was the experimentation based on a central composite design and second was the optimization of process conditions by using response surface methodology. Three process variables were evaluated at two levels: methanol/oil molar ratio, reaction temperature and amount of catalyst. A second-order model was obtained to predict yield of fatty acid methyl esters as a function of the three evaluated variables with high statistical significance (p-value=0.0146). The best combination of process variables are 14:1 methanol/oil molar ratio, 0.42 wt% of NaOH and 43 °C.