Hydrogen production of photosynthetic bacteria group (PSBG) was studied using corn stalk hydrolyzate as hydrogen production substrate. The effects of sulfuric acid concentration, hydrolysis pH values and hydrolysis temperature on hydrogen production were investigated. The optimum process conditions were as follows: the sulfuric acid concentration was 1%, hydrolysis pH value was 6.0 and the hydrolysis temperature was 110°C , respectively. Three hydrolyzate detoxification methods were compared. The method of calcium hydroxide obtained the optimal detoxification effect and the maximum hydrogen yield was 472 mL H2/ (L-medium).