Biological dihydrogen (H2) production is a promising alternative to the conventional nonrenewable energies. H2 production by dark fermentation is particularly studied regarding its independence from fossil fuel and electricity, its cultivation into bioreactors and the broad range of substrate that can be used. A major goal application of dark fermentation is the valuation of wastes. In this context, cheese whey wastes are particularly adapted, since rich in organic matter, abundant and low cost. A novel H2-producing strain was isolated and identify as a Clostridium beijerinckii strain based on morphological and physiological characteristics, and 16S rDNA sequencing. The optimum temperature and pH for H2 production was 40°C and pH 7, respectively. Substrate and stress tolerance tests showed that C. beijerinckii C.sp.1.3 could produce H2 from glucose, lactose, maltose, cellobiose, galacturonic acid, xylose, and sucrose, and to a lesser extent from starch, cellulose and glycerol. The H2 production potential from a mix of dairy industrial wastes of this strain has been evaluated. The highest productivity in batch for 5 g.L -1 of Total Organic Carbon (TOC) was 833.1 mL H2.L -1 .d -1 , with a yield of 2.03 mMH2.mMlactose -1 , demonstrating the potential of this strain for H2 production from industrial wastes.