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Hydrogen production by dark fermentation is one promising technology. However, there are challenges in improving the performance and efficiency of the process. The important factors that must be considered to obtain a suitable process are the source of the inoculum and its pre-treatment, types of substrates, the reactor configurations and the hydrogen partial pressure. Furthermore, to obtain high-quality hydrogen, it is necessary to integrate an effective separation procedure that is compatible with the intrinsic characteristics of a biological process. Recent studies have suggested that a stable and robust process could be established if there was an effective selection of a mixed microbial consortium with metabolic pathways directly targeted to high hydrogen yields. Additionally, the integration of membrane technology for the extraction and separation of the hydrogen produced has advantages for the upgrading step, because this technology could play an important role in reducing the negative effect of the hydrogen partial pressure. Using this technology, it has been possible to implement a production-purification system, the 'hydrogen-extractive membrane bioreactor'. This configuration has great potential for direct applications, such as fuel cells, but studies of new membrane materials, module designs and reactor configurations are required to achieve higher separation efficiencies.

Many wild type Pseudomonas strains have the potential to contribute to the valorization of lignin in future biorefineries. Through a robust aromatic catabolism, i.e., biofunneling capacity, they can ease the inherent aromatic heterogeneity found in lignin hydrolysates and accumulate naturally marketable biopolymers like mcl-polyhydroxyalkanoate (mcl-PHA) under nitrogen limitation. Besides a comparative strain evaluation, we present fundamental research on the funneling of aromatic mixtures under specific bioprocess conditions to improve biocatalytic lignin valorization. For the most robust and best performing strain, P. putida KT2440, we improve the mcl-PHA accumulation from a defined aromatic mixture of p-coumarate, ferulate, and benzoate under technically relevant conditions by up to 40% by tailoring the nitrogen and oxygen supply. The highest mcl-PHA concentration (582 ± 41 mg L–1) was obtained for a C/N ratio of 60 for oxygen-unlimited conditions (oxygen transfer rate >20 mmol L–1 h–1). In contrast, aromatic intermediates accumulated under oxygen-limited conditions at oxygen transfer rates below 10 mmol L–1 h–1. The experimental conditions were scalable into a 1L stirred tank bioreactor. This study contributes to deepening our understanding of the biocatalytic capability of promising Pseudomonas strains toward downstream microbial conversions of lignin aromatics for future biorefinery applications.