Anaerobic sequencing batch reactor (ASBR) process offers great potential for H-2 production from wastewaters. In this study, an ASBR was used at first time for enhanced continuous H-2 production from fungal pretreated cornstalk hydrolysate by Thermoanaerobacterium thermosaccharolyticum W16. The reactor was operated at different hydraulic retention times (HRTs) of 6, 12, 18, and 24 h by keeping the influent hydrolysate constant at 65 mmol sugars L-1. Results showed that increasing the HRT from 6 to 12 h led to the H-2 production rate increased from 6.7 to the maximum of 9.6 mmol H-2 L-1 h(-1) and the substrate conversion reached 90.3%, although the H-2 yield remained at the same level of 1.7 mol H-2 mol(-1) substrate. Taking into account both H-2 production and substrate utilization efficiencies, the optimum HRT for continuous H-2 production via an ASBR was determined at 12 h. Compared with other continuous H-2 production processes, ASBR yield higher H2 production at relatively lower HRT. ASBR is shown to be another promising process for continuous fermentative H-2 production from lignocellulosic biomass. Copyright (C) 2014, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.