Three-stage process for tequila vinasse valorization through sequential lactate, biohydrogen and methane production.

This study evaluated a novel three-stage process devoted to the cascade production of lactate, biohydrogen and methane from tequila vinasse (TV), with emphasis on attaining a high and stable biohydrogen production rate (HPR) by utilizing lactate as biohydrogen precursor. In the first stage, tailored operating conditions applied to a sequencing batch reactor were effective in sustaining a lactate concentration of 12.4 g/L, corresponding to 89% of the total organic acids produced. In the second stage, the stimulation of lactate-centered dark fermentation which entails the decoupling of biohydrogen production from carbohydrates utilization was an effective approach enabling stable biohydrogen production, having HPR fluctuations less than 10% with a maximum HPR of 12.3 L/L-d and a biohydrogen yield of 3.1 L/LTV. Finally, 1.6 L CH4/L-d and 6.5 L CH4/LTV were obtained when feeding the biohydrogen fermentation effluent to a third methanogenic stage, yielding a global energy recovery of 267.5 kJ/LTV.

Changes in performance and bacterial communities in a continuous biohydrogen-producing reactor subjected to substrate- and pH-induced perturbations.

The performance and microbial communities of a continuous dark fermentation reactor exposed to perturbations induced by substrate change and acidic pH shock were investigated. A mesophilic well-mixed reactor separately fed with two types of tequila vinasses (TVs) and lactose was operated at a fixed pH of 5.5, except during short-term pH (3.8) stress, for ~61 days at decreasing hydraulic retention times (HRTs) from 24 to 4 h. During the first ~23 days of operation with TV, a decrease in HRT down to 4 h resulted in the highest reported biohydrogen productivity from TV of 12.4 NL/L-d. It was shown that abrupt change in TV type (even with temporal feeding of lactose) and transient over-acidification impaired the normal operation of the reactor. However, it rapidly recovered from such disturbances, sustaining similar high-rate productivity to that previously encountered. Recovery was attributed to resistant and resilient microbial community features, as supported by molecular characterisation.

Effect of process parameters on enhanced biohydrogen production from tequila vinasse via the lactate-acetate pathway.

In this study, a lactate-type fermentation entailing the consumption of lactate and acetate (lactate-acetate pathway) is proposed to deal with lactic acid bacteria (LAB) inhibition during the production of biohydrogen (bioH2) from tequila vinasse. The effects of total solids content, substrate concentration, nutrient formulation and inoculum addition on bioH2 production performance were investigated. Batch experiments were performed in a 3-L completely mixed reactor at 35 °C and pH 6.5-5.8. The lactate-acetate pathway mediated consistent bioH2 production which was influenced by inoculum addition followed by substrate concentration, nutrient formulation and solids content. Maximum bioH2 production rate (225 NmL/L-h) and yield (124 NmL/g VSadded) were achieved by removing suspended solids and enhancing nutrient content, respectively. Illumina sequencing-based analysis revealed a dominance of Clostridium in the inoculum, which together with LAB and acetic acid bacteria shaped a keystone cluster for avoiding LAB inhibition while ensuring consistent bioH2 production performance.