1AbstractBackgroundProduction of alcohols from wastes through biological processes is environmentally and economically interesting, since they can be valorized as drop-in liquid fuels, which have a high market value. Using microbial mixed cultures in such processes is of great interest since it confers more stability, a higher resistance to both toxicity and contamination, and an increased substrate flexibility. However, there is still a lack of fundamental knowledge on such microbial populations used as inoculum in solventogenic processes. This work evaluates the effect of four different physicochemical pretreatments (acidic, thermal, acidic-thermal and thermal-acidic) on an anaerobic inoculum used for alcohols production from volatile fatty acids.ResultsAll experiments were conducted in single batches using acetate and butyrate as substrates, at 30°C and with a pressurized headspace of pure H2 at 2182 mBar. Higher productions of both ethanol and butanol were achieved with both thermal and acidic-thermal pretreatments of the inoculum. The highest concentrations of ethanol and butanol produced were respectively of 122 mg.L−1 and 97 mg.L−1 for the thermal pretreatment (after 710 hours), and of 87 mg.L−1 and 143 mg.L−1 for the acidic-thermal pretreatment (after 210 hours). Butyrate was consumed and acetate was produced in all assays. A mass balance study indicated that the inoculum provided part of the substrate. Thermodynamic data indicated that a high H2 partial pressure favored solventogenic metabolic pathways. Finally, sequencing data showed that both thermal and acidic-thermal pretreatments selected mainly the bacterial genera Pseudomonas, Brevundimonas and Clostridium.ConclusionThe acidic-thermal pretreatment selected a bacterial community more adapted to the conversion of acetate and butyrate into ethanol and butanol, respectively. A higher production of ethanol was achieved with the thermal pretreatment, but at a slower rate. The thermal-acidic pretreatment was unstable, showing a huge variability between replicates. The acidic pretreatment showed the lowest alcohol production, almost negligible as compared to the control assay.