Dilute sulphuric acid pretreatment followed by enzyme saccharification of Sorghum bicolor straw was undertaken to examine its potential as a feedstock in bioethanol production in Australia. Factorial design experiments evaluated the impact of pretreatment parameters on hemicellulose solubilisation and cellulose enzymatic hydrolysis. Sugar yields in prehydrolysate and saccharified liquors were found to increase with treatment severity; temperature was found to have the greatest impact. Degradation products were minimal; acetate and total phenolics peaked at 33 and 1.5 mg/g respectively. Conditions for maximum hemicellulose solubilisation (2% H2SO4 for 60 min at 121 degrees C) differed to those associated with maximum glucose release from solid residue saccharifications (1% H2SO4/90 min /121 degrees C). Water extractive sugars accounted for over 20% total sugars recovered. Addition of beta-glucosidase and xylanase to enzyme saccharification enhanced reaction rates and final sugar yields three-fold, whilst reducing cellulase dosage. Considering its abundance, high sugar potential and apparent ease of conversion, sorghum straw is an appropriate feedstock for the production of second generation fuels. Crown Copyright (C) 2011 Published by Elsevier Ltd. All rights reserved.