Stalk of corn (Zea mays L.), a commonly available crop residue in tropical and subtropical countries and found in plenty. In this study, we demonstrated that the pretreated corn stalk waste could be used to produce hydrogen. The isolated Bacillus licheniformis MSU AGM 2 strain (HM214759) from the pretreated paper mill effluent produced hydrogen under optimized conditions: carbon source (1 g/l), nitrogen source (12.5 g/l), temperature (35 degrees C) and pH (6.0). Alkaline pretreatment with 2% NaOH removed lignin by 48% from the corn stalk waste. Pretreated corn stalk ranges from 1 to 5 g/l were tested for the effective bacterial growth and hydrogen production. Kinetic parameters analyzed in 1 l bioreactor showed the maximum hydrogen production and hydrogen yield with 185 ml/l and 82.5 ml/g substrate, respectively. Growth profile and modified Gompertz model at the above mentioned condition fitted well (R-2, 0.93). Hence the anaerobic fermentation by the isolated strain had increased the hydrogen evolution rate with formic acid, acetate and butyrate concentrations at the end of the fermentation. Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.