Dark anaerobic fermentation is an interesting alternative method for producing biohydrogen (H-2) as a renewable fuel because of its low cost and various usable organic substrates. Pulping sludge from wastewater treatment containing plentiful cellulosic substrate could be feasibly utilized for H-2 production by dark fermentation. The objective of this study was to investigate the optimal proportion of pulping sludge to paper waste, the optimal initial pH, and the optimal ratio of carbon and nitrogen (C/N) for H-2 production by anaerobic seed sludge pretreated with heat. The pulping sludge was pretreated with NaOH solution at high temperature and further hydrolyzed with crude cellulase. Pretreatment of the pulping sludge with 3% NaOH solution under autoclave at 121 degrees C for 2 h, hydrolysis with 5 FPU crude cellulase at 50 degrees C, and pH 4.8 for 24 h provided the highest reducing sugar production yield (229.68+/-2.09 mg/g(TVS)). An initial pH of 6 and a C/N ratio of 40 were optimal conditions for H-2 production. Moreover, the supplement of paper waste in the pulping sludge enhanced the cumulative H-2 production yield. The continuous hydrogen production was further conducted in a glass reactor with nylon pieces as supporting media and the maximum hydrogen production yield was 151.70 ml/g(TVS).