Bioaugmentation strategies based on bacterial and methanogenic cultures to relieve stress in anaerobic digestion of protein-rich substrates

Agostini, S.; Bucci, L.; Doni, D.; Costantini, P.; Gupte, A.; Muller, B.; Sibilla, F.; Basaglia, M.; Casella, S.; Kougias, P. G.; Campanaro, S.; Favaro, L.; Treu, L.

doi:10.1101/2023.12.11.571062

bioRxiv · 10.1101/2023.12.11.571062

Bioaugmentation strategies based on bacterial and methanogenic cultures to relieve stress in anaerobic digestion of protein-rich substrates

Agostini, S.·Bucci, L.·Doni, D.·Costantini, P.·Gupte, A.·Muller, B.·Sibilla, F.·Basaglia, M.·Casella, S.·Kougias, P. G.·Campanaro, S.·Favaro, L.·Treu, L.

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Abstract

Anaerobic co-digestion of protein-rich substrates is a prominent strategy for converting valuable feedstocks into methane, but it releases ammonia, which can inhibit methanogenesis. This study developed a cutting-edge combined culturomic and metagenomic approach to investigate the microbial composition of an ammonia-tolerant biogas plant. Newly-isolated microorganisms were used for bioaugmentation of stressed batch reactors fed with casein, maize silage and their combination. A co-culture enriched with proteolytic bacteria was isolated, selected and compared with the proteolytic collection strain Pseudomonas lundensis DSM6252. The co-culture and P. lundensis were combined with the ammonia-resistant archaeon Methanoculleus bourgensis MS2 to boost process stability. A microbial population pre-adapted to casein was also tested for evaluating the digestion of protein-rich feedstock. The promising results suggest combining proteolytic bacteria and M. bourgensis could exploit microbial co-cultures to improve anaerobic digestion stability and ensure stable productivity even under the harshest of ammonia conditions. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=84 SRC="FIGDIR/small/571062v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@623848org.highwire.dtl.DTLVardef@5d2311org.highwire.dtl.DTLVardef@a95f70org.highwire.dtl.DTLVardef@1b8beb6_HPS_FORMAT_FIGEXP M_FIG C_FIG Highlights- High ammonia release from protein-rich substrates inhibits anaerobic digestion - Newly isolated bacterial strains from anaerobic digester were obtained - Proper bioaugmentation alleviates stress in casein and maize silage co-digestion - Bioaugmentation with a hydrolytic/hydrogenotrophic co-culture enhances CH4 yields

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Agostini, S., Bucci, L., Doni, D., Costantini, P., Gupte, A., Muller, B., Sibilla, F., Basaglia, M., Casella, S., Kougias, P. G., Campanaro, S., Favaro, L., Treu, L.. 2023-12-11. Bioaugmentation strategies based on bacterial and methanogenic cultures to relieve stress in anaerobic digestion of protein-rich substrates. https://doi.org/10.1101/2023.12.11.571062

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