This study addressed the utilization of an agro-waste, corn stover, as a renewable lignocellulosic feedstock for the fermentative H(2) production by the moderate thermophile Thermoanaerobacterium thermosaccharolyticum W16. The corn stover was first hydrolyzed by cellulase with supplementation of xylanase after delignification with 2% NaOH. It produced reducing sugar at a yield of 11.2 g L(-1) glucose, 3.4 g L(-1) xylose and 0.5 g L(-1) arabinose under the optimum condition of cellulase dosage 25 U g(-1) substrate with supplement xylanase 30 U g(-1) substrate. The hydrolyzed corn stover was sequentially introduced to fermentation by strain W16, where, the cell density and the maximum H(2) production rate was comparable to that on simulated medium, which has the same concentration of reducing sugars with hydrolysate. The present results suggest a promising combined hydrogen production process from corn stover with enzymatic hydrolysis stage and fermentation stage using W16. Crown Copyright (C) 2009 Published by Elsevier Ltd on behalf of Professor T. Nejat Veziroglu. All rights reserved.