Microbial electrosynthesis is an uprising concept for the combined carbon dioxide reduction and electricity storage in the form of green chemical compounds. Although several proof of principle studies show great promise, mass-transfer limitations of substrates, protons and products remains one of the issues that needs to be addressed to bring the systems towards greater scale applications. A previously tested solution formed force flow-through catholyte recirculation, but this set-up encountered difficulties with gas accumulation during start-up at higher current densities (∼ −10 kA/m ³), creating the need for a bypass to release gas. In this study, start-up at high current density was achieved without a bypass by using an alternating flow-through regime. This regime decreased the operating energy input from 221 to 136 kWh per kg of produced hydrogen and reached acetate production within 10 days after start-up at high current density and elongation to n-caproate after 45 days. Mass-transfer studies were included by microsensor measurements of local conditions (hydrogen concentration, pH) combined with thermodynamic calculations at the start and end of 60-days biotic experiments. The microorganisms on the cathode decreased pH gradients and consumed the formed hydrogen. The presence of Clostridium sensu stricto 12 and Peptococcaceae species were related to chain elongation activity, and the presence of Methanobrevibacter was linked to methanogenesis activity. By identifying the effects of different flow-through strategies on local concentrations and functional microbial groups, this work provides insights on the optimal conditions for microbial CO ₂ conversion and highlight the application potential of microbial electrosynthesis.

It has been reported that lipid droplets (LDs), called oleosomes, have an inherent ability to inflate or shrink when absorbing or fueling lipids in the cells, showing that their phospholipid/protein membrane is dilatable. This property is not that common for membranes stabilizing oil droplets and when well understood, it could be exploited for the design of responsive and metastable droplets. To investigate the nature of the dilatable properties of the oleosomes, we extracted them from rapeseeds to obtain an oil-in-water emulsion. Initially, we added an excess of rapeseed oil in the dispersion and applied high-pressure homogenization, resulting in a stable oil-in-water emulsion, showing the ability of the molecules on the oleosome membrane to rearrange and reach a new equilibrium when more surface was available. To confirm the rearrangement of the phospholipids on the droplet surface, we used molecular dynamics simulations and showed that the fatty acids of the phospholipids are solubilized in the oil core and are homogeneously spread on the liquid-like membrane, avoiding clustering with neighbouring phospholipids. The weak lateral interactions on the oleosome membrane were also confirmed experimentally, using interfacial rheology. Finally, to investigate whether the weak lateral interactions on the oleosome membrane can be used to have a triggered change of conformation by an external force, we placed the oleosomes on a solid hydrophobic surface and found that they destabilise, allowing the oil to leak out, probably due to a reorganisation of the membrane phospholipids after their interaction with the hydrophobic surface. The weak lateral interactions on the LD membrane and their triggered destabilisation present a unique property that can be used for a targeted release in foods, pharmaceuticals and cosmetics.

We developed a technique based on the use of microsensors to measure pH and H₂ gradients during microbial electrosynthesis. The use of 3D electrodes in (bio)electrochemical systems likely results in the occurrence of gradients from the bulk conditions into the electrode. Since these gradients, e.g., with respect to pH and reactant/product concentrations determine the performance of the electrode, it is essential to be able to accurately measure them. Apart from these parameters, also local oxidation-reduction potential and electric field potential were determined in the electrolyte and throughout the 3D porous electrodes. Key was the realization that the presence of an electric field disturbed the measurements obtained by the potentiometric type of microsensor. To overcome the interference on the pH measure, a method was validated where the signal was corrected for the local electric field measured with the electric potential microsensor. The developed method provides a useful tool for studies about electrode design, reactor engineering, measuring gradients in electroactive biofilms, and flow dynamics in and around 3D porous electrodes of (bio)electrochemical systems.

In microbial electrosynthesis (MES), microorganisms grow on a cathode electrode as a biofilm, or in the catholyte as planktonic biomass, and utilize CO₂for their growth and metabolism. Modification of the cathode with metals can improve MES performance, due to their catalytic activity for H2 production, which can be consumed by microorganisms, or via modifying the cathode properties. On the other hand, metals can have an inhibiting effect on MES. While these single roles of metals and their oxides have been identified, an investigation of the simultaneous effects on MES is still lacking. Here, we modify activated carbon (AC) electrodes with nickel (Ni) at high (5%) and low (0.01%) loadings, to investigate its combined effects on MES. Upon Ni impregnation, multiple factors explained the MES performance, including electrocatalytic H2 production, trace element availability, metal toxicity, Ni leaching and redeposition/bio-crystalization. Instead, the electrode surface properties (i.e., surface area and pore structure) were not affected by Ni addition. Compared to unmodified AC, low Ni loading did not improve abiotic H2 production, whereas at high Ni loading a 6-fold increase was observed. During biological experiments, low Ni loading resulted in over a 3-fold increase of acetate production and 35% higher planktonic growth, compared to unmodified AC. Instead, high Ni loading resulted in 25-fold increase of acetate production, 21% decrease of planktonic growth, and improved biofilm growth. Unmodified AC, and low and high Ni loading each resulted in unique microbial community composition. The effect of Ni on MES is therefore concentration-dependent, with apparently different mechanisms of interaction being prevalent at low or high Ni loadings.

Carbon dioxide (CO₂) can be converted to valuable products using different catalysts, including metal or biological catalysts (e. g. microorganisms). Some products formed by metal electrocatalysts can be further utilized by microorganisms, and therefore catalytic cooperation can be envisioned. To prevent cumbersome separations, it is beneficial when both catalyst work under the same conditions, or at least in the same reaction medium. Here, we will show that a formate-producing copper electrocatalyst can function in a biological medium. Furthermore, we will show that the effluent of the copper-containing reactor can be used without purification as the sole medium for a bio-reactor, inoculated with a mixed culture of microorganisms. In that second reactor, formate, H₂ and CO₂ are consumed by the microorganisms, forming acetate and methane. Compared to simple buffer electrolyte, catalytic activity of copper was improved in the presence of microbial growth medium, likely due to EDTA (Ethylenediaminetetraacetic acid) present in the latter.