Decreasing hydrogen partial pressure can not only increase the activity of the hydrogen enzyme but also decrease the products inhibition, so it is an appropriate method to enhance the fermentative hydrogen production from anaerobic mixed culture. The effect of biogas release method on anaerobic fermentative hydrogen production in batch culture system was compared, i.e., Owen method with intermediately release, continuous releasing method, and continuous releasing+ CO₂ absorbing. The experimental results showed that, at 35°C, initial pH 7.0 and glucose concentration of 10 g·L^-1, the hydrogen production was only 28 mL when releasing gas by Owen method, while it increased two times when releasing the biogas continuously. The cumulative hydrogen production could reach 155 mL when carbon dioxide in the gas stream was continuously absorbed by 1 mol·L^-1 NaOH. The results showed that acetate was dominated, accounting for 43% in the dissolved fermentation products in Owen method, whereas the butyrate predominated and reached 47%–53% of the total liquid end products when releasing gas continuously. It is concluded that the homoacetogenesis could be suppressed when absorbing CO₂ in the gas phase in fermentative hydrogen production system.