In this study, the corn stover was pretreated, enzymatically hydrolyzed, and then fermented in the lipid production fermentation. The corn stover fermentation effluent was utilized for the photo-fermentation of a Rhodobacter sphaeroides ZX-5 for hydrogen production. The hydrogen production was more than twofolds greater than that in the synthetic medium under the similar organic acid concentration range. The synergism among the pure organic acids was found to facilitate cell growth and hydrogen production, although some organic acid was not utilized for hydrogen production directly. The synergism among the components in the corn stover fermentation effluent was also found. The initial pH value was found to be an important parameter for the photo-fermentation of R. sphaeroide ZX-5 using the corn stover fermentation effluent. The results provided a possible way to utilize lignocellulose-derived organic acids for hydrogen production, and to treat fermentation wastewater in biofuel production using lignocellulose. (C) 2010 Elsevier Ltd. All rights reserved.