Biodiesel is a reliable and promising replacement of fossil diesel. It is stable, less toxic and can be produced from sustainable resources, including a variety of raw materials. Currently, the most widely used are vegetable oils (edible and nonedible), due to their availability. The present paper considers the potential of obtaining fatty acid ethyl esters (FAEE) from corn oil, which is a byproduct of bioethanol production process. The ultimate outcome would definitely increase the profitability of the initial bioethanol production process. The biodiesel production process was implemented in two steps, due to the high content of free fatty acids of the obtained corn oil. The first step includes an acid-catalyzed esterification process and the second step comprises an alkali-catalyzed transesterification process to receive FAEE. Two different catalysts (sulfuric acid and p-toluene sulfonic acid) were utilized and compared each other in the esterification process, in order to cope with high acid number of the raw material. A comprehensive qualitative and quantitative analysis of both feedstock and biodiesel was performed using gas chromatography-mass spectrometry method. The obtained biodiesel was characterized by a significantly lower cloud point compared to the feedstock and high acid number.