Electrochemically active biofilms: facts and fiction. A review

This review examines the electrochemical techniques used to study extracellular electron transfer in the electrochemically active biofilms that are used in microbial fuel cells and other bioelectrochemical systems. Electrochemically active biofilms are defined as biofilms that exchange electrons with conductive surfaces: electrodes. Following the electrochemical conventions, and recognizing that electrodes can be considered reactants in these bioelectrochemical processes, biofilms that deliver electrons to the biofilm electrode are called anodic, ie electrode-reducing, biofilms, while biofilms that accept electrons from the biofilm electrode are called cathodic, ie electrode-oxidizing, biofilms. How to grow these electrochemically active biofilms in bioelectrochemical systems is discussed and also the critical choices made in the experimental setup that affect the experimental results. The reactor configurations used in bioelectrochemical systems research are also described and the authors demonstrate how to use selected voltammetric techniques to study extracellular electron transfer in bioelectrochemical systems. Finally, some critical concerns with the proposed electron transfer mechanisms in bioelectrochemical systems are addressed together with the prospects of bioelectrochemical systems as energy-converting and energy-harvesting devices.     

Electrochemically active biofilms: facts and fiction. A review

This review examines the electrochemical techniques used to study extracellular electron transfer in the electrochemically active biofilms that are used in microbial fuel cells and other bioelectrochemical systems. Electrochemically active biofilms are defined as biofilms that exchange electrons with conductive surfaces: electrodes. Following the electrochemical conventions, and recognizing that electrodes can be considered reactants in these bioelectrochemical processes, biofilms that deliver electrons to the biofilm electrode are called anodic, ie electrode-reducing, biofilms, while biofilms that accept electrons from the biofilm electrode are called cathodic, ie electrode-oxidizing, biofilms. How to grow these electrochemically active biofilms in bioelectrochemical systems is discussed and also the critical choices made in the experimental setup that affect the experimental results. The reactor configurations used in bioelectrochemical systems research are also described and the authors demonstrate how to use selected voltammetric techniques to study extracellular electron transfer in bioelectrochemical systems. Finally, some critical concerns with the proposed electron transfer mechanisms in bioelectrochemical systems are addressed together with the prospects of bioelectrochemical systems as energy-converting and energy-harvesting devices.     

Exocellular electron transfer in anaerobic microbial communities

Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory syntrophic consortia of proton-reducing acetogenic bacteria and hydrogen-consuming methanogenic archaea. Anaerobic microorganisms that use insoluble electron acceptors for growth, such as iron- and manganese-oxide as well as inert graphite electrodes in microbial fuel cells, also transfer electrons exocellularly. Soluble compounds, like humic substances, quinones, phenazines and riboflavin, can function as exocellular electron mediators enhancing this type of anaerobic respiration. However, direct electron transfer by cell-cell contact is important as well. This review addresses the mechanisms of exocellular electron transfer in anaerobic microbial communities. There are fundamental differences but also similarities between electron transfer to another microorganism or to an insoluble electron acceptor. The physical separation of the electron donor and electron acceptor metabolism allows energy conservation in compounds as methane and hydrogen or as electricity. Furthermore, this separation is essential in the donation or acceptance of electrons in some environmental technological processes, e.g. soil remediation, wastewater purification and corrosion.     

黄素介导的胞外电子转移研究与工程改造*

产电微生物的胞外电子转移在能源、环境等诸多领域有着非常重要的应用价值。希瓦氏菌(Shewanella oneidensis)作为模式产电微生物,其电催化系统引起了广泛的研究。黄素作为S. oneidensis重要的电子载体,其介导的胞外电子转移是电子传递过程中的一个限速步骤。然而自然环境中野生型S. oneidensis的黄素分泌量极低,对其工程改造也存在一定的局限性,因而严重阻碍了胞外电子的传递过程,这已成为限制其电子转移的主要瓶颈。基于S. oneidensis黄素介导的电子转移机制,系统地从黄素的合成路径及转录调控的角度阐明了黄素合成的调控因素,并综述近年来利用代谢工程、合成生物学以及电极材料修饰等方法来提高黄素介导电子转移的工程化策略,未来可利用一些系统的研究方法和表达工具来加速产电微生物黄素介导的胞外电子转移。     

Microbial extracellular electron transfer and its relevance to iron corrosion

Extracellular electron transfer (EET) is a microbial metabolism that enables efficient electron transfer between microbial cells and extracellular solid materials. Microorganisms harbouring EET abilities have received considerable attention for their various biotechnological applications, including bioleaching and bioelectrochemical systems. On the other hand, recent research revealed that microbial EET potentially induces corrosion of iron structures. It has been well known that corrosion of iron occurring under anoxic conditions is mostly caused by microbial activities, which is termed as microbiologically influenced corrosion (MIC). Among diverse MIC mechanisms, microbial EET activity that enhances corrosion via direct uptake of electrons from metallic iron, specifically termed as electrical MIC (EMIC), has been regarded as one of the major causative factors. The EMIC‐inducing microorganisms initially identified were certain sulfate‐reducing bacteria and methanogenic archaea isolated from marine environments. Subsequently, abilities to induce EMIC were also demonstrated in diverse anaerobic microorganisms in freshwater environments and oil fields, including acetogenic bacteria and nitrate‐reducing bacteria. Abilities of EET and EMIC are now regarded as microbial traits more widespread among diverse microbial clades than was thought previously. In this review, basic understandings of microbial EET and recent progresses in the EMIC research are introduced.     

电活性微生物胞外聚合物的特征与应用

电活性微生物是一类能够通过直接接触、导电菌毛或氧化还原介质与电极或者其他细胞进行胞外电子传递的微生物。而在这个过程中,胞外聚合物(extracellular polymeric substances, EPS)扮演着重要的角色。EPS是微生物生长过程中通过细胞裂解、水解分泌的高分子聚合物的混合物,主要由蛋白质、多糖和腐殖质等物质组成。来自电活性微生物的EPS的不同组成成分和特性会对EPS的电活性以及电活性微生物胞外电子传递产生一定的影响,同时在环境应用方面发挥重要作用。因此,为了更全面了解电活性微生物EPS的电活性及其对电活性微生物胞外电子传递的作用,本文总体介绍了电活性微生物EPS的电活性的直接表征方法,再从组成成分、化学性质、物理性质和空间分布4个方面综述了其对EPS电活性的影响及其在电子传递中的作用,介绍了当前电活性微生物EPS在染料废水脱色、重金属吸附、有机污染物的生物转化和渗滤液管理等方面的环境应用,并从表征方法、试验规模和互作机理研究等角度展望了未来的研究方向。     

Electrochemical characterization of Geobacter sulfurreducens cells immobilized on graphite paper electrodes

Bacteria able to transfer electrons to conductive surfaces are of interest as catalysts in microbial fuel cells, as well as in bioprocessing, bioremediation, and corrosion. New procedures for immobilization of Geobacter sulfurreducens on graphite electrodes are described that allow routine, repeatable electrochemical analysis of cell-electrode interactions. Immediately after immobilizing G. sulfurreducens on electrodes, electrical current was obtained without addition of exogenous electron shuttles or electroactive polymers. Voltammetry and impedance analysis of pectin-immobilized bacteria transferring electrons to electrode surfaces could also be performed. Cyclic voltammetry of immobilized cells revealed voltage-dependent catalytic current similar to what is commonly observed with adsorbed enzymes, with catalytic waves centered at -0.15 V (vs. SHE). Electrodes maintained at +0.25 V (vs. SHE) initially produced 0.52 A/m(2) in the presence of acetate as the electron donor. Electrical Impedance Spectroscopy of coatings was also consistent with a catalytic mechanism, controlled by charge transfer rate. When electrodes were maintained at an oxidizing potential for 24 h, electron transfer to electrodes increased to 1.75 A/m(2). These observations of electron transfer by pectin-entrapped G. sulfurreducens appear to reflect native mechanisms used for respiration. The ability of washed G. sulfurreducens cells to immediately produce electrical current was consistent with the external surface of this bacterium possessing a pathway linking oxidative metabolism to extracellular electron transfer. This electrochemical activity of pectin-immobilized bacteria illustrates a strategy for preparation of catalytic electrodes and study of Geobacter under defined conditions.     

Bacterial extracellular electron transfer in bioelectrochemical systems

Bioelectrochemical systems (BES), typically microbial fuel cells (MFCs), have attracted increasing attention in the past decade due to their promising applications in many fields, such as bioremediation, energy generation and biosynthesis. Current-generating microorganisms play a key role in BES. The process of transferring electrons to electrode has been considered as a novel anaerobic bacteria respiration, and more and more bacteria capable of exchanging electrons with electrodes have been isolated. Among those bacteria, Shewanella and Geobacter genera are the most frequently used model organisms in the studies of BES, as well as the bacteria-electrode electron transfer mechanisms. Many significant new findings in the field of the bacterial extracellular electron transfer in BES have been reported recently. A better understanding of the mechanisms of bacterial extracellular electron transfer would provide more efficient strategies to enhance the applicability of BES. This review summarizes the recent advances of extracellular electron transfer mechanisms with foci on Shewanella and Geobacter species in BES.     

Bioelectrochemical fuel cells

In the past 20 years, inorganic fuel cells have been transformed from novelty devices to practical energy transfer-energy storage units. However, the advantage of the high operating efficiency afforded by these fuel cells is partially offset by (a) the limited viability and high cost of the catalysts, (b) the highly corrosive electrolytes, and (c) the elevated operating temperatures. The possibility exists to reduce some of these problems through the development of bioelectrochemical fuel cells. Such biological/electrochemical systems incorporate either microorganisms or enzymes as an active component within the specified electrode compartments. Recent studies with microorganisms as part of the anode compartment have been aimed at defining the mechanism of the observed electrochemical reactions. Recent investigations on the use of cell-free enzyme preparations in the electrode compartments have dealt primarily with developing methodology and defining mechanisms for enhancing the rate of electron transfer from the enzyme-cofactor active site to the solid electrode surface. Applications of this developing technology have been envisioned for analytical chemistry, medical devices, energy transfer, electrochemical synthesis, and detoxification. In this review, the theory and problems of bioelectrochemical fuel cells are described and related to research, both recent and proposed, for the practical development of this area.     

Improved electrical wiring of microbes: anthraquinone-modified electrodes for biosensing of chlorinated hydrocarbons

The present study reports the development of a novel bioelectrochemical sensor for trichloroethene (TCE), a common subsurface contaminant, based on the measurement of the electrical current resulting from the microbially catalysed reduction of TCE at anthraquinone (AQ)-modified electrodes. Firstly, we describe the development and electrochemical characterisation of AQ-modified electrodes, prepared via spontaneous or electrochemical reduction of AQ diazonium derivatives. Finally, the proof-of-principle of the bioelectrochemical sensor for TCE was evaluated, using a TCE-dechlorinating microbial culture as the biosensing element. The response of the bioelectrochemical sensor was measured either as the peak current in cyclic voltammetry or the steady-state current in chronoamperometry; in both cases, it was found to be proportional to TCE concentrations in the range 0-100 μmol/L. On the other hand, the microorganisms in contact with the electrode surface caused severe fouling problems which drastically reduced the life-time of the sensor.     

Electroactive bacteria—molecular mechanisms and genetic tools

In nature, different bacteria have evolved strategies to transfer electrons far beyond the cell surface. This electron transfer enables the use of these bacteria in bioelectrochemical systems (BES), such as microbial fuel cells (MFCs) and microbial electrosynthesis (MES). The main feature of electroactive bacteria (EAB) in these applications is the ability to transfer electrons from the microbial cell to an electrode or vice versa instead of the natural redox partner. In general, the application of electroactive organisms in BES offers the opportunity to develop efficient and sustainable processes for the production of energy as well as bulk and fine chemicals, respectively. This review describes and compares key microbiological features of different EAB. Furthermore, it focuses on achievements and future prospects of genetic manipulation for efficient strain development.     

Bucking the current trend in bioelectrochemical systems: a case for bioelectroanalytics

Turning wastewater directly into electricity is alluring, widespread use of microbial fuel cells (MFCs) to achieve this at industrial scale appears increasingly unlikely despite intense research efforts lasting over a decade. Such endeavors have not been futile, however, and game-changing discoveries have resulted from these well-intentioned, scientifically rigorous but ultimately frustrated attempts to resolve the Waste-Energy dichotomy. The appeal of MFCs is largely of conceptual elegance rather than financial competitiveness, based on the green ideal that bacteria can be turned into cost effective bio-batteries. This notion is founded on the solid principles of extracellular electron transfer (EET), where microbes use electrodes interchangeably with other electron acceptors to generate current as a direct proxy for microbial metabolism. We contend that a nuanced understanding of EET has been restricted by focusing on device performance when in fact this information could be more beneficially channeled into addressing analytical questions pertaining to the presence and activity of microorganisms across systems of environmental and medical import, i.e. bioelectroanalytics. We discuss here relevant literature detailing bioelectrochemical systems and contrast energy-centric conclusions with observations geared towards bioelectroanalytics. We explore the expanding possibilities of bioelectroanalytics enabled by advances in genetic techniques and rooted in the concept that microbial interactions with an electrode extend to more than just cells seeking alternative electron acceptors. Our intention is to highlight alternative directions in the field and encourage researchers to harness bioelectroanalytics to address wider societal problems, in addition to addressing climate change.     

Cathodes as electron donors for microbial metabolism: Which extracellular electron transfer mechanisms are involved?

This review illuminates extracellular electron transfer mechanisms that may be involved in microbial bioelectrochemical systems with biocathodes. Microbially-catalyzed cathodes are evolving for new bioprocessing applications for waste(water) treatment, carbon dioxide fixation, chemical product formation, or bioremediation. Extracellular electron transfer processes in biological anodes, were the electrode serves as electron acceptor, have been widely studied. However, for biological cathodes the question remains: what are the biochemical mechanisms for the extracellular electron transfer from a cathode (electron donor) to a microorganism? This question was approached by not only analysing the literature on biocathodes, but also by investigating known extracellular microbial oxidation reactions in environmental processes. Here, it is predicted that in direct electron transfer reactions, c-type cytochromes often together with hydrogenases play a critical role and that, in mediated electron transfer reactions, natural redox mediators, such as PQQ, will be involved in the bioelectrochemical reaction. These mechanisms are very similar to processes at the bioanode, but the components operate at different redox potentials. The biocatalyzed cathode reactions, thereby, are not necessarily energy conserving for the microorganism.     

Screening of natural phenazine producers for electroactivity in bioelectrochemical systems

Mediated extracellular electron transfer (EET) might be a great vehicle to connect microbial bioprocesses with electrochemical control in stirred-tank bioreactors. However, mediated electron transfer to date is not only much less efficient but also much less studied than microbial direct electron transfer to an anode. For example, despite the widespread capacity of pseudomonads to produce phenazine natural products, only Pseudomonas aeruginosa has been studied for its use of phenazines in bioelectrochemical applications. To provide a deeper understanding of the ecological potential for the bioelectrochemical exploitation of phenazines, we here investigated the potential electroactivity of over 100 putative diverse native phenazine producers and the performance within bioelectrochemical systems. Five species from the genera Pseudomonas, Streptomyces, Nocardiopsis, Brevibacterium and Burkholderia were identified as new electroactive bacteria. Electron discharge to the anode and electric current production correlated with the phenazine synthesis of Pseudomonas chlororaphis subsp. aurantiaca. Phenazine-1-carboxylic acid was the dominant molecule with a concentration of 86.1 μg/ml mediating an anodic current of 15.1 μA/cm². On the other hand, Nocardiopsis chromatogenes used a wider range of phenazines at low concentrations and likely yet-unknown redox compounds to mediate EET, achieving an anodic current of 9.5 μA/cm². Elucidating the energetic and metabolic usage of phenazines in these and other species might contribute to improving electron discharge and respiration. In the long run, this may enhance oxygen-limited bioproduction of value-added compounds based on mediated EET mechanisms.     

微生物种间直接电子传递机理及应用研究进展

微生物胞内产生的电子转移到其他电子受体而获得能量的过程称为微生物胞外电子传递,其中,另一微生物作为电子受体时发生的电子传递称为微生物种间电子传递。根据微生物种间电子传递机制,可分间接种间电子传递和种间直接电子传递。由于种间直接电子传递不需要其他物质介导,因此较间接种间电子传递效率更高、能量利用更高。本文系统阐述了微生物进行胞外电子传递的机理及应用,重点分析了种间直接电子传递机理,并概述种间直接电子传递应用领域,为寻找更多电连接的微生物群落以及应用微生物提供参考。     

Electrochemically mediated CO<Subscript>2</Subscript> reduction for bio-methane production: a review

A number of methods for carbon capture, more specifically, CO₂ capture have been researched in the past few years. One such method is electrochemical CO₂ reduction to biomethane which also serves the purpose of biogas upgradation using microbial electrosynthesis systems. This technology is also known as Power to Gas technology and the review starts with the importance and requirement of PtG in the modern world by studying energy production and consumption patterns in Europe, with a focus on Norway. The paper summarises the recent works and concepts in the field of bioelectrochemical systems with a focus on electron transfer mechanisms, biocatalysts and reactor designs. Works and gaps in the studies of direct interspecies electron transfer and biocathode developments are discussed in detail. This is followed by a discussion explaining various reactor designs, the advantages of single chambered microbial reactors and the importance of reactors that combine anaerobic digestion with microbial electrolysis cells.     

Application of redox mediators in bioelectrochemical systems

Redox mediators (RM) are natural or artificial compounds used by microorganisms as electron acceptors and electron donors during electron transfer. Evidence collected in the last years indicates that the application of RM in bioelectrochemical systems (BES) enhanced the electron transfer from microorganisms to anodes and from cathodes to microorganisms. This review summarizes the results of using soluble or immobilized RM in BES to produce electricity and for the treatment of contaminants from wastewater effluents. In addition, future research focused on biohydrogen production, recovery or removal metals, and the use of humic substances (HS) extracted from natural environment is proposed.     

Electricity generation by microorganisms in the sediment-water interface of an extreme acidic microcosm

The attachment of microorganisms to electrodes is of great interest for electricity generation in microbial fuel cells (MFC) or other applications in bioelectrochemical systems (BES). In this work, a microcosm of the acidic ecosystem of Río Tinto was built and graphite electrodes were introduced at different points. This allowed the study of electricity generation in the sediment/water interface and the involvement of acidophilic microorganisms as biocatalysts of the anodic and cathodic reactions in a fuel-cell configuration. Current densities and power outputs of up to 3.5 A/m2 and 0.3 W/m2, respectively, were measured at pH 3. Microbial analyses of the electrode surfaces showed that Acidiphilium spp., which uses organic compounds as electron donors, were the predominant biocatalysts of the anodic reactions, whereas the aerobic iron oxidizers Acidithiobacillus ferrooxidans and Leptospirillum spp. were detected mainly on the cathode surface.     

微生物燃料电池在毒性物质检测中的应用

微生物燃料电池(Microbial fuel cell,MFC)利用微生物整体作为催化剂催化底物将化学能直接转化为电能,是一种极具应用前景的生物电化学技术。微生物在阳极氧化还原有机物产生电子并传递给阳极,电子通过外电路传递至阴极后将电子释放给阴极中的氧化剂,从而产生电流。当有毒物质进入MFC,微生物活性降低,电子传递量变少,电流降低,而电流的产生与微生物活性呈线性关系,据此可检测样品的毒性。本文主要介绍了微生物燃料电池在毒性物质抗生素、重金属离子、有机污染物、酸等方面的研究,并分析了微生物燃料电池存在的问题及未来研究方向,以期不久的将来微生物燃料电池能付之使用。     

分子生物学方法提高电活性微生物胞外电子传递效率的研究进展*

微生物细胞与电极之间的胞外电子传递效率是限制微生物电化学技术发展的关键因素,而分子生物学的发展为提高胞外电子传递效率带来了光明前景。从四种具有代表性的纯培养电活性微生物(奥奈达希瓦氏菌、铜绿假单胞菌、硫还原地杆菌和工程大肠杆菌)和混合培养电活性微生物出发,综述了利用分子生物学手段改造几种电活性微生物的研究成果,阐明了针对特异的电活性微生物,如何采取相应的分子生物学手段提高其胞外电子传递的效率,并展望了未来的研究方向。