Novel insights on the versatility of biohydrogen production from sugarcane vinasse via thermophilic dark fermentation: Impacts of pH-driven operating strategies on acidogenesis metabolite profiles

An innovative application of the anaerobic structured-bed reactor (AnSTBR) in thermophilic dark fermentation of sugarcane vinasse targeting biohydrogen (bioH₂) production was assessed. A detailed metabolite monitoring program identified the major substrates and primary metabolic pathways within the system. Increasing the applied organic loading rate positively affected bioH₂ production, reaching 2074 N mL-H₂ L^-1 d^-1 and indicating an optimal load of approximately 70 kg-COD m^-3 d^-1. Controlling the fermentation pH (5.0-5.5) was the primary strategy to maintain bioH₂-producing conditions, offsetting negative impacts associated with the compositional variability of vinasse. Metabolic correlations pointed out lactate as the primary substrate for bioH₂ production, indicating its accumulation as evidence of impaired reactors. The versatility of the acidogenic system was confirmed by identifying three major metabolic pathways according to the pH, i.e., lactate-producing (pH <5.0), bioH₂-/butyrate-producing (pH = 5.0-5.5) and bioH₂-producing/sulfate-reducing (pH >6.0) systems, which enables managing the operation of the reactors for diversified purposes in practical aspects.