In this study the starch from Solanum lycocarpum fruit was used for bioethanol production. S. lycocarpum starch was hydrolyzed in a serial reactor containing α-amylase and glucoamylase immobilized onto glutaraldehyde-derivative polyaniline, producing 91.6% of glucose for a 10% starch suspension and 86.3% of glucose for a 20% starch suspension after 60 min of reaction. Fermentation parameters for bioethanol production were evaluated by using a full factorial design (3²) with fermentation time (h) and concentration of glucose equivalent (%) as independent variables. Results from factorial design showed that the highest ethanol production was found when 1% of glucose equivalent was fermented by 48 h. At these conditions the bioethanol conversion efficiency reached 92.13% and it was estimated a productivity of 596.96 L tonne⁻¹. It can be concluded that the starch from S. lycocarpum fruit is a promising non-food and renewable energy source for bioethanol production.