For improving photo-biohydrogen production, a novel gas bubble column photobioreactor with Ar gas sparging was developed for biohydrogen production by purple non-sulfur phototrophic bacteria, Rhodoseudomonas palustris CQK-01. The dissolved hydrogen concentration was in-situ measured by a hydrogen microsensor. Experimental results demonstrated that Ar gas sparging dramatically decreased the dissolved hydrogen concentration, resulting in an improvement in the photo-biohydrogen production performance. Furthermore, effects of the gas flow rate and the time interval of gas sparging were investigated. The results showed that with an increase in the gas flow rate, the hydrogen production performance increased initially due to the reduced dissolved hydrogen concentration and enhanced mass transport, and then it decreased as a result of an increased shear stress. Meanwhile, the short sparging time interval resulted in a low accumulation of dissolved hydrogen in the bioreactor, hence high hydrogen production performance. The optimal hydrogen production rate (5.86 mmol/l/h) and hydrogen yield (3.38 mol H-2/mol glucose) were obtained at the gas flow rate of 10 ml/min, respectively. Copyright (C) 2012, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.