Effect of dilution and L-malic acid addition on bio-hydrogen production with Rhodopseudomonas palustris from effluent of an acidogenic anaerobic reactor

In this study, H2 was produced in a two-stage biological process: I) first stage; the dark fermentation of cheese whey wastewater, which is rich in lactose, by mixed anaerobic culture grown at thermophilic temperature in a continuously running fermentor and ii) second stage; the photo-fermentation of the residual medium by R. palustris strain (DSM 127) at 31oC under illumination of 150 W in batch mode, respectively. In the first part of the study, the effluent from the dark fermentation reactor was used either as it is (no dilution) or after dilution with distilled water at varying ratios such as 1/2 , 1/5, 1/10 (1 volume effluent/5 volume distilled water) before used in photo-fermentation experiments. In the second part of the study, L-malic acid at varying amounts was added into the hydrogen production medium in order to have L-malic acid concentrations ranging from 0 to 4 g/l. Non-diluted and pre-diluted mediums with or without L-malic acid addition were also tested for comparison purpose (as controls). Prior to the hydrogen production experiments, all samples were subjected to pH adjustment, (pH 6.7) and sterilized by autoclave at 121oC for 15 min. In regards to the experiments in which the effect of dilution of the effluent from dark fermentation was studied, it was observed that dilution of the effluent from dark fermentation resulted in much better hydrogen productions. Among the dilution rates used, the experiments operated with 1/5 dilution ratio produced the best hydrogen production (241 ml H2/ g CODfed). On the other hand, it was seen that the mixing the effluent with L-malic acid (0 - 4 g/l) at increasing ratios (studied from 0% L-malic acid up to 100% by volume in the mixture) had further positive effect and improved the hydrogen production. The bioreactors containing only L-malic acid media resulted in the best hydrogen production (438 ml H2 / g CODfed). It was found that, undiluted raw cheese whey wastewater effluent from dark hydrogen production reactor was not suitable for photo-fermentative hydrogen production. It was concluded that dilution of the feeding helps to reduce the nitrogen content and the volatile fatty acid content that might be otherwise harmful to the photo-heterotrophic organisms. The second conclusion that can be drawn is that cheese whey effluent should be mixed with L-malic acid rich co-substrates such as fruit juice processing effluents before fed into the photo-fermentation reactor. Finally, the two-stage H2-producing process could be applied in remediation of lactose-containing industrial wastes, H2 being used on-site, to reduce process costs via generation of electricity by the help of hydrogen fuel cells. (author)