Microbe Electrode Electron Transfer: Understanding Interactions from the Enzyme to the Microbial Community Level

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Microbial Biotechnology".

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 23449

Special Issue Editors


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Guest Editor
Institute of Applied Biosciences, Karlsruhe Institute of Technology (KIT),· 76131 Karlsruhe, Germany
Interests: anode- and cathode-assisted fermentation; electrode biofilms; extracellular respiration

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Guest Editor
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-156 Oeiras, Portugal
Interests: multiheme cytochromes; extracellular electron transfer; electroactive organisms; electron transfer mechanisms
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Special Issue Information

Dear Colleagues,

A fundamental understanding of the respiratory interaction of enzymes, individual cells, and biofilms with electrodes is key to elucidating new physiological principles but also to applying and developing microorganisms for microbial electrochemical technologies. We have recently seen that the field has been expanding rapidly with the discovery of new biocatalysts, new applications, new ways to accelerate electron transfer, and a better understanding of the fundamental biochemistry sustaining the microbe–electrode interaction. This Special Issue will compile studies on how organisms and biofilms interact with electrodes and how this understanding can be used to advance their application. Moreover, we will highlight future research questions and trends in the field of bioelectrochemistry.

Hence, with this Special Issue we encourage colleagues to submit papers that address: (1) the fundamental microbial biochemistry of extracellular electron transfer onto electrodes; (2) the impact of biofilms and their structure on electron transfer kinetics; (3) the characterization of new biocatalysts acting on the anode or cathode site; and (4) the further development of strains for electrode-assisted or driven processes.

Prof. Dr. Johannes Gescher
Dr. Catarina M. Paquete
Guest Editors

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Keywords

  • microbe–electrode electron transfer
  • microbial electrochemical technologies
  • biofilms
  • extracellular respiration

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Published Papers (6 papers)

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Research

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13 pages, 1589 KiB  
Article
Crossing the Wall: Characterization of the Multiheme Cytochromes Involved in the Extracellular Electron Transfer Pathway of Thermincola ferriacetica
by Marisa M. Faustino, Bruno M. Fonseca, Nazua L. Costa, Diana Lousa, Ricardo O. Louro and Catarina M. Paquete
Microorganisms 2021, 9(2), 293; https://doi.org/10.3390/microorganisms9020293 - 31 Jan 2021
Cited by 17 | Viewed by 3899
Abstract
Bioelectrochemical systems (BES) are emerging as a suite of versatile sustainable technologies to produce electricity and added-value compounds from renewable and carbon-neutral sources using electroactive organisms. The incomplete knowledge on the molecular processes that allow electroactive organisms to exchange electrons with electrodes has [...] Read more.
Bioelectrochemical systems (BES) are emerging as a suite of versatile sustainable technologies to produce electricity and added-value compounds from renewable and carbon-neutral sources using electroactive organisms. The incomplete knowledge on the molecular processes that allow electroactive organisms to exchange electrons with electrodes has prevented their real-world implementation. In this manuscript we investigate the extracellular electron transfer processes performed by the thermophilic Gram-positive bacteria belonging to the Thermincola genus, which were found to produce higher levels of current and tolerate higher temperatures in BES than mesophilic Gram-negative bacteria. In our study, three multiheme c-type cytochromes, Tfer_0070, Tfer_0075, and Tfer_1887, proposed to be involved in the extracellular electron transfer pathway of T. ferriacetica, were cloned and over-expressed in E. coli. Tfer_0070 (ImdcA) and Tfer_1887 (PdcA) were purified and biochemically characterized. The electrochemical characterization of these proteins supports a pathway of extracellular electron transfer via these two proteins. By contrast, Tfer_0075 (CwcA) could not be stabilized in solution, in agreement with its proposed insertion in the peptidoglycan wall. However, based on the homology with the outer-membrane cytochrome OmcS, a structural model for CwcA was developed, providing a molecular perspective into the mechanisms of electron transfer across the peptidoglycan layer in Thermincola. Full article
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15 pages, 8694 KiB  
Article
Improving the Cathodic Biofilm Growth Capabilities of Kyrpidia spormannii EA-1 by Undirected Mutagenesis
by Tobias Jung, Max Hackbarth, Harald Horn and Johannes Gescher
Microorganisms 2021, 9(1), 77; https://doi.org/10.3390/microorganisms9010077 - 30 Dec 2020
Cited by 11 | Viewed by 3024
Abstract
The biotechnological usage of carbon dioxide has become a relevant aim for future processes. Microbial electrosynthesis is a rather new technique to energize biological CO2 fixation with the advantage to establish a continuous process based on a cathodic biofilm that is supplied [...] Read more.
The biotechnological usage of carbon dioxide has become a relevant aim for future processes. Microbial electrosynthesis is a rather new technique to energize biological CO2 fixation with the advantage to establish a continuous process based on a cathodic biofilm that is supplied with renewable electrical energy as electron and energy source. In this study, the recently characterized cathodic biofilm forming microorganism Kyrpidia spormannii strain EA-1 was used in an adaptive laboratory evolution experiment to enhance its cathodic biofilm growth capabilities. At the end of the experiment, the adapted cathodic population exhibited an up to fourfold higher biofilm accumulation rate, as well as faster substratum coverage and a more uniform biofilm morphology compared to the progenitor strain. Genomic variant analysis revealed a genomically heterogeneous population with genetic variations occurring to various extends throughout the community. Via the conducted analysis we identified possible targets for future genetic engineering with the aim to further optimize cathodic growth. Moreover, the results assist in elucidating the underlying processes that enable cathodic biofilm formation. Full article
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15 pages, 2119 KiB  
Article
Coupling an Electroactive Pseudomonas putida KT2440 with Bioelectrochemical Rhamnolipid Production
by Theresia D. Askitosari, Carola Berger, Till Tiso, Falk Harnisch, Lars M. Blank and Miriam A. Rosenbaum
Microorganisms 2020, 8(12), 1959; https://doi.org/10.3390/microorganisms8121959 - 10 Dec 2020
Cited by 19 | Viewed by 3928
Abstract
Sufficient supply of oxygen is a major bottleneck in industrial biotechnological synthesis. One example is the heterologous production of rhamnolipids using Pseudomonas putida KT2440. Typically, the synthesis is accompanied by strong foam formation in the reactor vessel hampering the process. It is caused [...] Read more.
Sufficient supply of oxygen is a major bottleneck in industrial biotechnological synthesis. One example is the heterologous production of rhamnolipids using Pseudomonas putida KT2440. Typically, the synthesis is accompanied by strong foam formation in the reactor vessel hampering the process. It is caused by the extensive bubbling needed to sustain the high respirative oxygen demand in the presence of the produced surfactants. One way to reduce the oxygen requirement is to enable the cells to use the anode of a bioelectrochemical system (BES) as an alternative sink for their metabolically derived electrons. We here used a P. putida KT2440 strain that interacts with the anode using mediated extracellular electron transfer via intrinsically produced phenazines, to perform heterologous rhamnolipid production under oxygen limitation. The strain P. putida RL-PCA successfully produced 30.4 ± 4.7 mg/L mono-rhamnolipids together with 11.2 ± 0.8 mg/L of phenazine-1-carboxylic acid (PCA) in 500-mL benchtop BES reactors and 30.5 ± 0.5 mg/L rhamnolipids accompanied by 25.7 ± 8.0 mg/L PCA in electrode containing standard 1-L bioreactors. Hence, this study marks a first proof of concept to produce glycolipid surfactants in oxygen-limited BES with an industrially relevant strain. Full article
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16 pages, 1645 KiB  
Article
Impact of Inoculum Type on the Microbial Community and Power Performance of Urine-Fed Microbial Fuel Cells
by Maria Jose Salar-Garcia, Oluwatosin Obata, Halil Kurt, Kartik Chandran, John Greenman and Ioannis A. Ieropoulos
Microorganisms 2020, 8(12), 1921; https://doi.org/10.3390/microorganisms8121921 - 3 Dec 2020
Cited by 22 | Viewed by 3025
Abstract
Bacteria are the driving force of the microbial fuel cell (MFC) technology, which benefits from their natural ability to degrade organic matter and generate electricity. The development of an efficient anodic biofilm has a significant impact on the power performance of this technology [...] Read more.
Bacteria are the driving force of the microbial fuel cell (MFC) technology, which benefits from their natural ability to degrade organic matter and generate electricity. The development of an efficient anodic biofilm has a significant impact on the power performance of this technology so it is essential to understand the effects of the inoculum nature on the anodic bacterial diversity and establish its relationship with the power performance of the system. Thus, this work aims at analysing the impact of 3 different types of inoculum: (i) stored urine, (ii) sludge and (iii) effluent from a working MFC, on the microbial community of the anodic biofilm and therefore on the power performance of urine-fed ceramic MFCs. The results showed that MFCs inoculated with sludge outperformed the rest and reached a maximum power output of 40.38 mW·m−2anode (1.21 mW). The power performance of these systems increased over time whereas the power output by MFCs inoculated either with stored urine or effluent decreased after day 30. These results are directly related to the establishment and adaptation of the microbial community on the anode during the assay. Results showed the direct relationship between the bacterial community composition, originating from the different inocula, and power generation within the MFCs. Full article
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16 pages, 3648 KiB  
Article
Accelerated Electro-Fermentation of Acetoin in Escherichia coli by Identifying Physiological Limitations of the Electron Transfer Kinetics and the Central Metabolism
by Sebastian Beblawy, Laura-Alina Philipp and Johannes Gescher
Microorganisms 2020, 8(11), 1843; https://doi.org/10.3390/microorganisms8111843 - 23 Nov 2020
Cited by 3 | Viewed by 2987
Abstract
Anode-assisted fermentations offer the benefit of an anoxic fermentation routine that can be applied to produce end-products with an oxidation state independent from the substrate. The whole cell biocatalyst transfers the surplus of electrons to an electrode that can be used as a [...] Read more.
Anode-assisted fermentations offer the benefit of an anoxic fermentation routine that can be applied to produce end-products with an oxidation state independent from the substrate. The whole cell biocatalyst transfers the surplus of electrons to an electrode that can be used as a non-depletable electron acceptor. So far, anode-assisted fermentations were shown to provide high carbon efficiencies but low space-time yields. This study aimed at increasing space-time yields of an Escherichia coli-based anode-assisted fermentation of glucose to acetoin. The experiments build on an obligate respiratory strain, that was advanced using selective adaptation and targeted strain development. Several transfers under respiratory conditions led to point mutations in the pfl, aceF and rpoC gene. These mutations increased anoxic growth by three-fold. Furthermore, overexpression of genes encoding a synthetic electron transport chain to methylene blue increased the electron transfer rate by 2.45-fold. Overall, these measures and a medium optimization increased the space-time yield in an electrode-assisted fermentation by 3.6-fold. Full article
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Review

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22 pages, 1921 KiB  
Review
Microfluidic Microbial Bioelectrochemical Systems: An Integrated Investigation Platform for a More Fundamental Understanding of Electroactive Bacterial Biofilms
by Stéphane Pinck, Lucila Martínez Ostormujof, Sébastien Teychené and Benjamin Erable
Microorganisms 2020, 8(11), 1841; https://doi.org/10.3390/microorganisms8111841 - 23 Nov 2020
Cited by 21 | Viewed by 5637
Abstract
It is the ambition of many researchers to finally be able to close in on the fundamental, coupled phenomena that occur during the formation and expression of electrocatalytic activity in electroactive biofilms. It is because of this desire to understand that bioelectrochemical systems [...] Read more.
It is the ambition of many researchers to finally be able to close in on the fundamental, coupled phenomena that occur during the formation and expression of electrocatalytic activity in electroactive biofilms. It is because of this desire to understand that bioelectrochemical systems (BESs) have been miniaturized into microBES by taking advantage of the worldwide development of microfluidics. Microfluidics tools applied to bioelectrochemistry permit even more fundamental studies of interactions and coupled phenomena occurring at the microscale, thanks, in particular, to the concomitant combination of electroanalysis, spectroscopic analytical techniques and real-time microscopy that is now possible. The analytical microsystem is therefore much better suited to the monitoring, not only of electroactive biofilm formation but also of the expression and disentangling of extracellular electron transfer (EET) catalytic mechanisms. This article reviews the details of the configurations of microfluidic BESs designed for selected objectives and their microfabrication techniques. Because the aim is to manipulate microvolumes and due to the high modularity of the experimental systems, the interfacial conditions between electrodes and electrolytes are perfectly controlled in terms of physicochemistry (pH, nutrients, chemical effectors, etc.) and hydrodynamics (shear, material transport, etc.). Most of the theoretical advances have been obtained thanks to work carried out using models of electroactive bacteria monocultures, mainly to simplify biological investigation systems. However, a huge virgin field of investigation still remains to be explored by taking advantage of the capacities of microfluidic BESs regarding the complexity and interactions of mixed electroactive biofilms. Full article
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