Effect of hyperthermophilic pretreatment on methane and hydrogen production from garden waste under mesophilic and thermophilic conditions

Abstract Anaerobic digestion of garden waste was investigated in a two-stage process consisting of hyperthermophilic pretreatment followed by mesophilic or thermophilic fermentation. The greatest digestion performance was achieved when the substrates were first treated at 70°C for 3 days with no inoculation, and then mixed with inoculum (anaerobic sludge) and subjected to anaerobic digestion at 55°C. Under such conditions, the maximum methane and hydrogen yields from grass were 517 NmlCH4/kgVS and 52 NmlH2/kgVS, whereas the corresponding values for leaves were 421 NmlCH4/kgVS and 23 NmlH2/kgVS, and these figure were far greater than the yields obtained in experiments with no hyperthermophilic stage. A metagenomic analysis of hyperthermophilic environments revealed the appearance of thermophilic and hyperthermophilic bacteria showing hydrolytic activity against lignocellulosic materials, including Caldicellulosiruptor, Thermovenabulum, Thermoanaerobacter, Moorella, Tepimicrobium, Geobacillus and Thermobacillus at a genus level. A noticeable methane production in the hyperthermophilic stage could be linked to the presence of Methanothermobacter sp.