Abstract. Anaerobic digestion can be used for the treatment of POME to reduce organic content and generate some substances, such as volatile acids, ethanol, and various gasses. During anaerobic fermentation process, microorganism requires the presence of trace elements for growth to improve their performance. This research will be carried out as a study of the presence of trace elements, such as metals (Cu 2+ , Mn 2+ , and Mg 2+ ) that has significant effects on ethanol formation. Circulating Bed Reactor was used and operated in a batch system for 48 hours. Metal ions were screened and analyzed by using two-level factorial design method whether there is any correlation effect between the addition of Cu 2+ , Mn 2+ , and Mg 2+ and ethanol formation. Several parameters which consists of Total Volatile Acids (TVA), dissolved Chemical Oxygen Demand (CODs), Volatile Suspended Solid (VSS), pH, Dissolved Oxygen (DO), and ethanol were measured every sampling. Mn metal is proven statistically affect both TVA and ethanol formation while Mg metal only affect TVA formation. Cu 2+ and Mg 2+ metals combination affect ethanol formation with largest detected ethanol concentration is 7,483.07 mg/L. The result from this study had identified the metal ions which has significant effect as a foundation for optimization ethanol formation.