Auto selection of microorganisms of excess activated sludge used as an inoculum for fermentative hydrogen production from substrates with different biopolymeric composition

When mixed organic waste is used for hydrogen production by dark fermentation, the microbial community which is most adapted to actual biopolymer composition of the substrate, is auto-selected. In this research, 6 substrates simulating different biopolymers (proteins, fats, carbohydrates) and their mixtures were used to enrich hydrogenproducing bacteria adapted to these substrates from non-pretreated activated sludge. Phylum Firmicutes dominated in the microbial community (67-100%) regardless of the substrate used, as was shown by high-throughput sequencing. Microbial diversity was low when using carbohydrate-rich substrates, and microbial community was mainly represented by Ruminococcus (26-90%) and Thermoanaerobacterium (6-67%). Dark fermentation of fats and proteins was characterized by higher microbial diversity. Thermoanaerobacterium (21%), Thermobrachium (19%), Tepidiphilus (16%) and Acetomicrobium (14%) dominated when using fats, while Thermobrachium (34%), Acetomicrobium (16%) and Clostridium sensu stricto 7 (12%) dominated when using proteins as substrate. Different microbial communities and substrates resulted in diverse process performance and metabolic pathway. Dark fermentation of starch achieved the maximum hydrogen yield of 138 ml/g volatile solids with 60.4 % hydrogen content in biogas. The dominance of genus Ruminococcus was supposed to be responsible for the highest hydrogen production. Minor amounts of methane from proteins and fats were produced by Methanothermobacter and Methanosarcina. The hydrogenotrophic pathway was slightly more predominant methane formation route than the others considered based upon to the stable ¹³C isotope analysis.

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