Fast depletion of fossil fuels with high fluctuating market prices has made the hunt for alternate resources for the production of transportation fuels mandatory. Our approach is to utilize rice straw as feedstock for the production of alcoholic biofuels. In this paper, we have compared the effect of various pretreatment processes on rice straw, viz. physical (steam under pressure) and chemical (acid) and enzymatic treatments as a precursor to ABE fermentation for production of biobutanol. Glucose analysis was done by quantification of glucose content spectrophotometrically via Glucose assay kit. The rice straw hydrolyzate produced through pretreatment was allowed to undergo anaerobic fermentation, using C. acetobutylicum MTCC 481. The yield and productivity of ABE solvents (acetone, butanol and ethanol) was calculated using HPLC. The ABE yield produced using hydrolyzate obtained through enzyme assisted acid hydrolysis, viz. acetone: 0.11, butanol: 0.861, ethanol: 0.05, was found to be the best among all experiments. (C) 2011 Elsevier Ltd. All rights reserved.