Napier grass is a potential biomass for biofuel production due to its high adaptability andrnwidespread availability in tropical areas such as Ethiopia. A compositional analysis of Napierrngrass collected from the Menagesha Mushroom farm in an Addis Ababa city suburb wasrnperformed, and microbes and enzymes were purchased from France. An alkaline pretreatmentrn(2.5wt% NaOH, 1.5wt% urea, and 8wt% sample biomass) was mixed for 40 minutes at 121Â°C,rnfollowed by enzymatic hydrolysis (2ml Novozymes Cellic glucadase enzyme and 5gram pretreatedrnbiomass) shaken at 50rpm for 40 minutes at 50Â°C, and separated by centrifuge at 8500rpm tornhydrolyze Napier grass. Carbohydrates make up 52.1 % of the Napier grass composition, makingrnit an excellent carbon source. Acetone-Butanol-Ethanol fermentation (ABE) was carried out in arncontinuous fermentation process using co-cultured C. acetobutylicum bacteria. After optimizingrnthe operating parameters pH, Temperature, substrate concentration, and fermentation time, thernproductivity of butanol was found to be 0.077 g ltrn-1rn hrrn-1rn with a 29.5 % efficiency. The yield of ABErnwas 0.0833 g ltrn-1rn hrrn-1rn when compared to the semiâ€“Simultaneous Saccharification fermentationrn(sSSF) process, which combined alkaline pretreated biomass with enzymatic hydrolysis and coculturedrnbacteriarnafterrn24rnhoursrnofrnhydrolysisrnfollowedrnbyrn96hrsrnofrnfermentation.rnButanolrnefficiencyrnrnatrntherns-SSFrnprocessrnisrn33.6rn%.rnWhenrncomparedrntornpreviousrnstudies,rnABErnfermentationrnofrnNapierrnrngrassrnyieldsrna competitivernyield.