The feasibility of fungal pretreatment of cornstalk with Phanerochaete chrysosporium for enzymatic saccharification and H-2 production was investigated in this study. Firstly, cornstalk was pretreated with P. chtysosporium at 29 degrees C under static condition for 15 d, lignin reduction was up to 34.3% with holocellulose loss less than 10%. Microscopic structure observation combined FTIR analysis further demonstrated that the lignin and crystallinity were decreased. Subsequently, the fungal-pretreated cornstalk was subjected to enzymatic hydrolysis by the crude cellulase from Trichoderma viride to produce fermentable sugars which were then fermented to bio-H-2 using Thermoanaerobacterium thermosaccharolyticum W16. The maximum enzymatic saccharification was found to be 47.3% which was 20.3% higher than the control without pretreatment. Upon fermentation of enzymatic hydrolysate, the yield of H-2 was calculated to be 80.3 ml/g-pretreated cornstalk. The present results suggested the potential of using hydrogen-producing bacteria for high-yield conversion of cornstalk into bio-H-2 integrate with biological pretreatment and enzymatic saccharification. (C) 2012 Elsevier Ltd. All rights reserved.