In this study, production of hydrogen (H-2) from glucose, xylose, galactose, mannose, arabinose and rhamnose by a strain isolated from activated sludge was investigated. The strain, named as Citrobacter sp. CMC-1, was enriched in cellobiose amended minimal media. Based on 16S rRNA sequence, the CMC-1 strain is a close relative of Citrobacter amalonaticus strain SA01 (99%). Optimal cultivation parameters for H-2 production and growth such as pH and temperature were investigated. H-2 yields from glucose at optimal conditions (pH 6.0 and 34 degrees C) were 1.82 +/- 0.02 mol-H-2/mol-glucose. Strain CMC-1 fermented galactose, mannose, xylose, arabinose and rhamnose. After 48 h incubation, the strain CMC-1 completely fermented all sugars tested, except arabinose. Increase in fermentation period lowered residual formate level in the media and improved H-2 production for galactose, mannose and xylose (1.68 +/- 0.24, 1.93 +/- 0.14 and 1.63 +/- 0.07 mol-H-2/mol-substrate respectively). Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.