AbstractGrasses are abundant in many climatic regions of the world and have been regarded as weeds by many. This work investigated the use of Pennisetum purpureum (Napier grass) in the production of bioethanol. Two pretreated grasses were compared as the initial substance in the hydrolysis process followed by bacteria fermentation. For the purpose of breaking down lignin, alkali pretreatment, where grass was soaked in 7% NaOH, was used. For biological pretreatment, grass was incubated for 3 weeks with the white-rot fungus, Phanerochaete chrysosporium. Both types of pretreated materials were subjected to Trichoderma reesei ATCC 26921 enzyme hydrolysis. Glucose content from alkali-pretreated samples was 1.6-fold higher than fungus-pretreated samples. Hydrolysates from the pretreatments were fermented using the ethanol insensitive strain Escherichia coli K011. After 24 hours of fermentation, the ethanol yield from alkali-pretreated material was 1.5 times higher than the biological-pretreated material. It can be concluded that NaOH-pretreated enzyme hydrolysate had a better ethanol yield compared to biological-pretreated enzyme hydrolysate, but biological-pretreated enzyme hydrolysate had better ethanol conversion efficiency, which was 18.5 g/g. These results indicated that wild grass is capable of becoming an important biomass for small local bioethanol production.