Simultaneous Production of Biohydrogen (bioH₂) and Poly-Hydroxy-Alkanoates (PHAs) by a Photoheterotrophic Consortium Bioaugmented with Syntrophomonas wolfei

Mixed cultures represent better alternatives to ferment organic waste and dark fermentation products in anerobic conditions because the microbial associations contribute to electron transfer mechanisms and combine metabolic possibilities. The understanding of the microbial interactions in natural and synthetic consortia and the strategies to improve the performance of the processes by bioaugmentation provide insight into the physiology and ecology of the mixed cultures used for biotechnological purposes. Here, synthetic microbial communities were built from three hydrogen (bioH₂) and poly-hydroxy-alkanoates (PHA) producers, Clostridium pasteurianum, Rhodopseudomonas palustris and Syntrophomonas wolfei, and a photoheterotrophic mixed consortium C4, and their performance was evaluated during photofermentation. Higher hydrogen volumetric production rates (H₂VPR) were determined with the consortia (28–40 mL/Lh) as compared with individual strains (20–27 mL/Lh). The designed consortia reached the highest bioH₂ and PHA productions of 44.3 mmol and 50.46% and produced both metabolites simultaneously using dark fermentation effluents composed of a mixture of lactic, butyric, acetic, and propionic acids. When the mixed culture C4 was bioaugmented with S. wolfei, the bioH₂ and PHA production reached 32 mmol and 50%, respectively. Overall, the consumption of organic acids was above 50%, which accounted up to 55% of total chemical oxygen demand (COD) removed. Increased bioH₂ was observed in the condition when S. wolfei was added as the bioaugmentation agent, reaching up to 562 mL of H₂ produced per gram of COD. The enhanced production of bioH₂ and PHA can be explained by the metabolic interaction between the three selected strains, which likely include thermodynamic equilibrium, the assimilation of organic acids via beta-oxidation, and the production of bioH₂ using a proton driving force derived from reduced menaquinone or via electron bifurcation.