Biohydrogen production from synthetic wastewater as substrate was studied in anaerobic small scale batch reactors. Enriched anaerobic mixed consortia sampled from various environments were used as parent inocula to start the bioreactors. Selective enrichments were achieved by various physical and chemical pretreatments and changes in the microbial communities were monitored by metagenomic and molecular diagnostics approaches. Experimental data showed the feasibility of biohydrogen production using synthetic wastewater as substrate. The hydrogen generation capability of the different mixed consortia is clearly dependent on the pretreatment methods. The described approach opens the possibility for an alternative way towards simultaneous wastewater treatment and renewable energy generation. Copyright (C) 2013, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.