In the present study, the feasibility of combining dry dark fermentation and mechanical pretreatment of wheat straw was studied in order to improve substrate valorization, save energy input, decrease the environmental impact and diversify biofuels and volatile fatty acids production. To this end, dark fermentation of wheat straw was performed at 55 degrees C and 35 degrees C under solid-state conditions (23% of total solid content) and it was considered as a biological pretreatment. Both biologically treated and raw straws were reduced at four particles size to cover the range of fine (50 < X < 500 mu m) and ultrafine milling (<50 mu m). Biological pretreatment led to a substrate conversion of 16% and 14%, mainly into volatile fatty acids and biohydrogen. Biological pretreatment improved the substrate grindability with a reduction of mean particle size up to 31% and a reduction of the milling specific energy consumption up to 35% compared to untreated straw. Finally, related to untreated straw, this combination of biological and mechanical treatments improved the bioethanol yield up to 83%, which leads to an enhancement of the overall substrate conversion up to 131%. Based on these high yields, this combination of dry biological-mechanical pretreatments appears more attractive and efficient in terms of bioproducts production, energy efficiency and environmental impact, compared to conventional pretreatments. (C) 2015 Elsevier Ltd. All rights reserved.