Photo fermentation is a biological process that can be applied for hydrogen production. The process is environmental friendly which is operated under mild conditions using renewable resources. In order to increase yield of H2 produced by Rhodobacter sphaeroides, some experimental factors that may enhance H2 production were studied. The effect of operating parameters including agitation, aeration and light on hydrogen production using R. sphaeroides NCIMB 8253 was investigated. Rhodobacter sphaeroides NCIMB 8253 was grown in 100 mL serum bottle containing growth medium with maliec acid as the sole organic carbon source. The cultures were incubated anaerobically at 30 degrees C with tungsten lamp (100 W) as the light source (3.8 klux) and argon gas was purged for maintaining anaerobic condition. The results show that maximum hydrogen produced was higher (54.37 mL) in static culture with 69.98% of H2 in the total gas compared with shake culture (11.57 mL) with 57.86% of H2. By using static culture, H2 produced was five times higher compared with non-static in both aerobic and anaerobic condition. It was found that growth and H2 production with fluorescent lamp showed better results than growth and H2 production with tungsten light.
A two-stage fermentation process consisting of dark and photo-fermentation periods was carried out in a batch reactor. In the first stage, glucose was fermented in the dark stage using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564; CSN1-4) to produce acetate, CO2 and H2. The acetate produced in the first stage is fermented to H2 and CO2 by Rhodobacter sphaeroides NCIMB 8253 for further hydrogen production in the second, illuminated stage. The yield of hydrogen in the first stage was about 3.10 mol H2 (mol glucose)(-1) at a glucose concentration of 10 g L(-1), pH 6 +/- 0.2 and 37 degrees C and the second stage yield was about 1.10-1.25 mol H2 (mol acetic acid)(-1) at pH 6.8 +/- 0.2 and 32 degrees C, without removal of the Clostridium CSN1-4. The overall yield of hydrogen in the two-stage process, with glucose as the main substrate was higher than single-stage fermentation.