Electron bifurcation mechanism and homoacetogenesis explain products yields in mixed culture anaerobic fermentations

Anaerobic fermentation of organic wastes using microbial mixed cultures is a promising avenue to treat residues and obtain added-value products. However, the process has some important limitations that prevented so far any industrial application. One of the main issues is that we are not able to predict reliably the product spectrum (i.e. the stoichiometry of the process) because the complex microbial community behaviour is not completely understood. To address this issue, in this work we propose a new metabolic network of glucose fermentation by microbial mixed cultures that incorporates electron bifurcation and homoacetogenesis. Our methodology uses NADH balances to analyse published experimental data and evaluate the new stoichiometry proposed. Our results prove for the first time the inclusion of electron bifurcation in the metabolic network as a better description of the experimental results. Homoacetogenesis has been used to explain the discrepancies between observed and theoretically predicted yields of gaseous H2 and CO2 and it appears as the best solution among other options studied. Overall, this work supports the consideration of electron bifurcation as an important biochemical mechanism in microbial mixed cultures fermentations and underlines the importance of considering homoacetogenesis when analysing anaerobic fermentations

Performance and genome-centric metagenomics of thermophilic single and two-stage anaerobic digesters treating cheese wastes

The present research is the first comprehensive study regarding the thermophilic anaerobic degradation of cheese wastewater, which combines the evaluation of different reactor configurations (i.e. single and two-stage continuous stirred tank reactors) on the process efficiency and the in-depth characterization of the microbial community structure using genome-centric metagenomics. Both reactor configurations showed acidification problems under the tested organic loading rates (OLRs) of 3.6 and 2.4 g COD/L-reactor day and the hydraulic retention time (HRT) of 15 days. However, the two-stage design reached a methane yield equal to 95% of the theoretical value, in contrast with the single stage configuration, which reached a maximum of 33% of the theoretical methane yield. The metagenomic analysis identified 22 new population genomes and revealed that the microbial compositions between the two configurations were remarkably different, demonstrating a higher methanogenic biodiversity in the two-stage configuration. In fact, the acidogenic reactor of the serial configuration was almost solely composed by the lactose degrader Bifidobacterium crudilactis UC0001. The predictive functional analyses of the main population genomes highlighted specific metabolic pathways responsible for the AD process and the mechanisms of main intermediates production. Particularly, the acetate accumulation experienced by the single stage configuration was mainly correlated to the low abundant syntrophic acetate oxidizer Tepidanaerobacter acetatoxydans UC0018 and to the absence of aceticlastic methanogens

Cashew apple bagasse as new feedstock for the hydrogen production using dark fermentation process.

Cashew apple bagasse (CAB) has been studied as feedstock for the biohydrogen production using Clostridium roseum and the dark fermentation process. Pretreatment with alkaline hydrogen peroxide (CAB-AHP) on raw material and the acid and enzymatic hydrolysis have been taken into account to evaluate the H2 yields. Results show that the acid hydrolysate obtained from CAB produced higher H2 molar yield (HMY) (15 mmolH2/Lhydrolysate) than the acid hydrolysate from CAB-AHP (4.99 mmolH2/Lhydrolysate), These HMY were noticeably higher than values obtained from the enzymatic hydrolysate of CAB-AHP (1.05 mmolH2/Lhydrolysa) and the enzymatic hydrolysate of CAB (0.59 mmolH2/Lhydrolysa). The maximum biohydrogen productivity (12.57 mLH2/L.h) was achieved using the acid hydrolysate from CAB, with a H2 content of about 72% vol, that could be satisfactory in view of an energetic applications of the biogas. Results suggest that CAB could be considered for the hydrogen production process, providing an appropriate destination for this lignocellulosic biomass, and consequently, reducing the environmental impact it can exert