Thickness and roughness of transparent gold-palladium anodes have no impact on growth kinetics and yield coefficients of early-stage Geobacter sulfurreducens biofilms.

Geobacter sulfurreducens is the model organism for electroactive microorganisms performing direct extracellular electron transfer and forming thick mature biofilm electrodes. Although numerous physiological properties of mature biofilm electrodes are deciphered, there is an extensive gap of knowledge on the early-stage biofilm formation. We have shown recently that transparent gold-palladium (AuPd) electrodes allow for analysis of early-stage biofilm formation using confocal laser scanning microscopy. Here we analysed the influence of thickness (ranging from 12.5 to 200 nm) and roughness of AuPd electrodes on physiological parameters of G. sulfurreducens early-stage biofilms. We show that when grown potentiostatically at -200 mV vs. Ag/ AgCl sat. KCl neither maximum current density (jmax of âˆ¼ 80-150 µA cm-2) nor lag time (lag t of âˆ¼ 0.2-0.4 days) or single cell yield coefficients (YNe of 1.43 × 1012 cells mole--1) of the biofilms are influenced by the electrode preparation. This confirms the robustness of the experimental approach, which is an inevitable prerequisite for obtaining reliable results in follow-up experiments.

The electrode potential determines the yield coefficients of early-stage Geobacter sulfurreducens biofilm anodes.

Geobacter sulfurreducens is the model for electroactive microorganisms (EAM). EAM can use solid state terminal electron acceptors (TEA) including anodes via extracellular electron transfer (EET). Yield coefficients relate the produced cell number or biomass to the oxidized substrate or the reduced TEA. These data are not yet sufficiently available for EAM growing at anodes. Thus, this study provides information about kinetics as well as yield coefficients of early-stage G. sulfurreducens biofilms using anodes as TEA at the potentials of -200 mV, 0 mV and +200 mV (vs. Ag/AgCl sat. KCl). The selected microorganism was therefore cultivated in single and double chamber batch reactors on graphite or AuPd anodes. Interestingly, whereas the lag time and maximum current density within 12 days of growth differed, the anode potential does not influence the coulombic efficiency and the formal potential of the EET, which remains constant for all the experiments at ~ -300 to -350 mV. We demonstrated for the first time that the anode potential has a strong influence on single cell yield coefficients which ranged from 2.69 × 1012 cells mole--1 at -200 mV and 1.48 × 1012 cells mole--1 at 0 mV to 2.58 × 1011 cells mole--1 at +200 mV in single chamber reactors and from 1.15 × 1012 cells mole--1 at -200 mV to 8.98× 1011 cells mole--1 at 0 mV in double chamber reactors. This data can be useful for optimization and scaling-up of primary microbial electrochemical technologies.

Quantification of outer membrane vesicles: a potential tool to compare response in Pseudomonas putida KT2440 to stress caused by alkanols.

The bacterial release of outer membrane vesicles (OMVs) is an important physiological mechanism of Gram-negative bacteria playing numerous key roles. One function of the release of OMVs is related to an increase in surface hydrophobicity. This phenomenon initiates biofilm formation, making bacteria more tolerant to environmental stressors. Recently, it was qualitatively shown for Pseudomonas putida that vesicle formation plays a crucial role in multiple stress responses. Yet, no quantification of OMVs for certain stress scenarios has been conducted. In this study, it is shown that the quantification of OMVs can serve as a simple and feasible tool, which allows a comparison of vesicle yields for different experimental setups, cell densities, and environmental stressors. Moreover, the obtained results provide insight to the underlying mechanism of vesicle formation as it was observed that n-alkanols, with a chain length of C7 and longer, caused a distinct and steep increase in vesiculation (12-19-fold), compared to shorter chain n-alkanols (2-4-fold increase).