In this work, the optimization of hydrogen production by photo-fermentation bacteria immobilized on agar gel granule was systematic investigated in batch culture. Experiment focus on the effect of some important affecting factors on photo-hydrogen production. Results indicated that immobilized Rhodopseudomonas faecalis RLD-53 exhibited the highest hydrogen yield of 3.15 mol H2/mol acetate under follow optimal condition: agar granule diameter of 2.5 mm, inoculum age of 24 h, agar concentration of 2%, biomass of 4 mg/ml in agar and light intensity of 9000 lux. More importantly, immobilized photo-fermentation bacteria not only can enhance hydrogen production but can increase acids-tolerance capacity, even at pH 5.0 hydrogen also was produced, and thus hopefully immobilized photo-fermentation bacteria can be applied in the combination of dark and photo-fermentation for hydrogen production with high yield.
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