Cell number of Clostridium butyricum and Rhodobacter sphaeroides in co-culture was measured using q-PCR approach. During efficient H2 photoproduction from starch (6.2 mol H2/mol glucose), Clostridia growth and starch-hydrolyzing activity was partly suppressed. Apparently, the effect of R. sphaeroides towards C. butyricum was not attributed to altered Eh or pH values in the presence of purple bacteria. Further, disk-diffusion test proved that R. sphaeroides was capable of producing inhibitors against another purple bacterium, Rhodospirillum rubrum, but not against C. butyricum. We suggested that at initial cell number ratio C. butyricum:R. sphaeroides 1:1 purple bacteria outcompeted C. butyricum for yeast extract at its low concentration (80 mg/L). Under these conditions, the H2 yield was rather high (5.7 mol/mol). When the yeast extract concentration increased to 320 mg/L, this process was replaced by the low-yield H2 production (1.8 mol/mol) characteristic of Clostridia. However, increased percentage of purple bacteria in inoculum under these conditions prevented this shift. The outcome of competition depended on both the yeast extract concentration and cell number ratio. Apparently, the competition for yeast extract helped to maintain balance between fast-growing C. butyricum and slower-growing R. sphaeroides for efficient H2 photoproduction.
Keywords: Clostridia; Co-culture; Competition; Growth inhibition; Purple bacteria.