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Pb-contaminated soil poses inherent risks for the spread of antibiotic resistance genes (ARGs). However, few reports have investigated the distribution of lead resistance genes (LRGs), particularly their role in ARGs dynamics during Pb-contaminated soil remediation. This study explored the mechanisms driving ARGs variation under different remediation strategies. The results indicated that an increase in the total abundance of ARGs after applying montmorillonite (Imvite), the composite remediation agents of MgO and Ca(H2PO4)2 (MgO-Ca(H2PO4)2), and the composite remediation agents of montmorillonite, MgO, and Ca(H2PO4)2 (Imvite-MgO-Ca(H2PO4)2). Bioelectrochemical systems (BES) effectively reduced ARGs abundance, and when combined with Imvite-MgO-Ca(H2PO4)2, lowered the risk of ARGs proliferation linked to antibiotic deactivation. Changes in Pb concentration and pH reshaped microbial communities, impacting both LRGs and ARGs. To reduce the risk of ARGs proliferation and transmission during, various control strategies, such as modifying Pb stress, adjusting pH, and manipulating microbial community structure, have been proposed. The study provided theoretical foundation and practical strategies for controlling ARGs dissemination during the remediation of heavy metal-contaminated soil.

Plastic wastes, widely distributed in the environment, can be transformed into microplastics, posing a huge threat to ecosystems and human health due to their stability and adsorbability to other toxic pollutants (e.g., heavy metals and antibiotics). Recently, microbial degradation of (micro)plastics has gained widespread attention because of its green and sustainable properties. Microbial degradation of (micro)plastics is based on the cascade effects of various enzymes secreted by microorganisms, which can convert (micro)plastics into oligomers and monomers, or even mineralize them into CO2 and H2O. The microbial degradation of (micro)plastics is affected by multiple factors, such as microbial species, plastic properties, and environmental conditions. Currently, limited efficient plastic-degrading microorganisms have been discovered, and their degradation mechanisms are still unclear. Furthermore, the efficiency of microbial degradation needs to be improved for future application. Therefore, this review systematically summarizes the sources and properties of existing plastics, identifies pure cultures and mixed cultures for plastic degradation, and examines their influencing factors. In particular, the microbial degradation behaviors of (micro)plastics, including relevant enzymes, degradation efficiency, and degradation mechanisms, were thoroughly discussed. Additionally, the augmentation technologies coupling with microbial degradation, such as advanced oxidation, electrochemical, and genetic engineering technologies, were introduced and highlighted for their potential prospects. This review provides a reference for future research and development of (micro)plastic biodegradation technology.

Bioelectrochemical systems (BESs) are an innovative technology for the efficient degradation of antibiotics. Shewanella oneidensis (S. oneidensis) MR-1 plays a pivotal role in degrading sulfamethoxazole (SMX) in BESs. Our study investigated the effect of BES conditions on SMX degradation, focusing on microbial activity. The results revealed that BESs operating with a 0.05 M electrolyte concentration and 2 mA/cm2 current density outperformed electrolysis cells (ECs). Additionally, higher electrolyte concentrations and elevated current density reduced SMX degradation efficiency. The presence of nutrients had minimal effect on the growth of S. oneidensis MR-1 in BESs; it indicates that S. oneidensis MR-1 can degrade SMX without nutrients in a short period of time. We also highlighted the significance of mass transfer between the cathode and anode. Limiting mass transfer at a 10 cm electrode distance enhanced S. oneidensis MR-1 activity and BES performance. In summary, this study reveals the complex interaction of factors affecting the efficiency of BES degradation of antibiotics and provides support for environmental pollution control.

The over-exploitation of fossil fuels and their negative environmental impacts have attracted the attention of researchers worldwide, and efforts have been made to propose alternatives for the production of sustainable and clean energy. One proposed alternative is the implementation of bioelectrochemical systems (BESs), such as microbial fuel cells (MFCs), which are sustainable and environmentally friendly. MFCs are devices that use bacterial activity to break down organic matter while generating sustainable electricity. Furthermore, MFCs can produce bioelectricity from various substrates, including domestic wastewater (DWW), municipal wastewater (MWW), and potato and fruit wastes, reducing environmental contamination and decreasing energy consumption and treatment costs. This review focuses on recent advancements regarding the design, configuration, and operation mode of MFCs, as well as their capacity to produce bioelectricity (e.g., 2203 mW/m2) and fuels (i.e., H2: 438.7 mg/L and CH4: 358.7 mg/L). Furthermore, this review highlights practical applications, challenges, and the life-cycle assessment (LCA) of MFCs. Despite the promising biotechnological development of MFCs, great efforts should be made to implement them in a real-time and commercially viable manner.

The effect of bioelectrochemical anaerobic digestion (BEAD) on the methanogenic performance of brewery wastewater at different organic loading rates (OLRs) was investigated and compared to conventional anaerobic digestion. A continuous BEAD reactor was used to treat brewery wastewater at different OLRs of 2, 4, 8, 16, and 20 g COD/L.d. The experimental results showed that the methane production was gradually increased from 0.48 L/L.d at an OLR of 2 g COD/L.d to 5.64 L/L.d at an OLR of 20 g COD/L.d. The methane production of the BEAD system was significantly higher than that of the conventional anaerobic reactor, indicating that BEAD has a better treatment effect for brewery wastewater. The performance of the conventional anaerobic reactor was significantly reduced especially at an OLR of 16 g COD/L.d, while the BEAD system could withstand a higher OLR. Bioelectrochemical systems provide a completely new platform for the anaerobic treatment of brewery wastewater and greatly improve the operation of anaerobic processes.

Rumen fermentation (RF) is a complex system of oxidation–reduction reactions governed by the rumen microbiota, and its end-products are the volatile fatty acids (VFAs) which constitute the main energy source for the host animal. We speculated that the process could be manipulated through electrofermentation (EF) by utilizing solid-state electrodes as electron sources. This study aimed to evaluate the effects of single-chamber EF applied (0.4 vs. 0.9 V) for 6, 12, or 24 h to rumen fluid on the resulting microbial biomass, dry matter digestibility (DMD), VFA production, NADH/NAD ratio, and composition of the rumen fluid’s bacterial communities. The application of a 0.9 V electric potential for 24 h produced the most significant changes in RF compared to the control treatment (0V), increasing microbial biomass (12%), DMD (58%), and VFA concentrations (~84%), and reducing NAD/NADH ratio (~50%) without a significant impact on bacterial diversity. These results suggest that the application of a 0.9 V electric potential allows the microbiota to better hydrolyze and digest the feed’s components and could offer an electron sink for the production of VFAs. Hence, EF can provide innovative solutions to a range of challenges that limit the RF process. Nonetheless, more studies are needed to understand how the solid electrodes and external electric energy function.

A microbial fuel cell (MFC) is a bioelectrochemical system that can be employed for the generation of electrical energy under microbial activity during wastewater treatment practices. The optimization of electrode spacing is perhaps key to enhancing the performance of an MFC. In this study, electrode spacing was evaluated to determine its effect on the performance of MFCs. The experimental work was conducted utilizing batch digesters with electrode spacings of 2.0 cm, 4.0 cm, 6.0 cm, and 8.0 cm. The results demonstrate that the performance of the MFC improved when the electrode spacing increased from 2.0 to 6.0 cm. However, the efficiency decreased after 6.0 cm. The digester with an electrode spacing of 6.0 cm enhanced the efficiency of the MFC, which led to smaller internal resistance and greater biogas production of 662.4 mL/g VSfed. The electrochemical efficiency analysis demonstrated higher coulombic efficiency (68.7%) and electrical conductivity (177.9 µS/cm) for the 6.0 cm, which was evident from the enrichment of electrochemically active microorganisms. With regards to toxic contaminant removal, the same digester also performed well, revealing removals of over 83% for chemical oxygen demand (COD), total solids (TS), total suspended solids (TSS), and volatile solids (VS). Therefore, these results indicate that electrode spacing is a factor affecting the performance of an MFC, with an electrode spacing of 6.0 cm revealing the greatest potential to maximize biogas generation and the degradability of wastewater biochemical matter.

Ammonia removal from wastewater was successfully achieved by simultaneous nitrification and denitrification (SND) in a double-chamber microbial electrolysis cell (MEC). The MEC operations at different applied voltages (0.7 to 1.5 V) and initial ammonia concentrations (30 to 150 mg/L) were conducted in order to evaluate their effects on MEC performance in batch mode. The maximum nitrification efficiency of 96.8% was obtained in the anode at 1.5 V, followed by 94.11% at 1.0 V and 87.05% at 0.7. At 1.5 V, the initial ammonia concentration considerably affected the nitrification rate, and the highest nitrification rate constant of 0.1601/h was determined from a first-order linear regression at 30 mg/L ammonium nitrogen. The overall total nitrogen removal efficiency was noted to be 85% via the SND in the MEC operated at an initial ammonium concentration of 50 mg/L and an applied cell voltage of 1.5 V. The MEC operation in continuous mode could remove ammonia (50 mg/L) in a series of anode and cathode chambers at the nitrogen removal rate of 170 g-N/m3.d at an HRT of 15. This study suggests that a standalone dual-chamber MEC can efficiently remove ammonia via the SND process without needing additional organic substrate and aeration, which makes this system viable for field applications.

A milk-processing plant was drafted as a distinctive staple industry amid the diverse field of industries. Dairy products such as yogurt, cheese, milk powder, etc., consume a huge amount of water not only for product processing, but also for sanitary purposes and for washing dairy-based industrial gear. Henceforth, the wastewater released after the above-mentioned operations comprises a greater concentration of nutrients, chemical oxygen demand, biochemical oxygen demand, total suspended solids, and organic and inorganic contents that can pose severe ecological issues if not managed effectively. The well-known processes such as coagulation–flocculation, membrane technologies, electrocoagulation, and other biological processes such as use of a sequencing batch reactor, upflow sludge anaerobic blanket reactor, etc., that are exploited for the treatment of dairy effluent are extremely energy-exhaustive and acquire huge costs in terms of fabrication and maintenance. In addition, these processes are not competent in totally removing various contaminants that exist in dairy effluent. Accordingly, to decrease the energy need, microbial electrochemical technologies (METs) can be effectively employed, thereby also compensating the purification charges by converting the chemical energy present in impurities into bioelectricity and value-added products. Based on this, the current review article illuminates the application of diverse METs as a suitable substitute for traditional technology for treating dairy wastewater. Additionally, several hindrances on the way to real-world application and techno-economic assessment of revolutionary METs are also deliberated.

Bioelectrochemical systems are emerging technologies for the reduction in CO2 in fuels and chemicals, in which anaerobic chemoautotrophic microorganisms such as methanogens and acetogens are typically used as biocatalysts. The anaerobic digestion digestate represents an abundant source of methanogens and acetogens microorganisms. In a mixed culture environment, methanogen’s inhibition is necessary to avoid acetate consumption by the presence of acetoclastic methanogens. In this study, a methanogenesis inhibition approach based on the thermal treatment of mixed cultures was adopted and evaluated in terms of acetate production under different tests consisting of hydrogenophilic and bioelectrochemical experiments. Batch experiments were carried out under hydrogenophilic and bioelectrochemical conditions, demonstrating the effectiveness of the thermal treatment and showing a 30 times higher acetate production with respect to the raw anaerobic digestate. Moreover, a continuous flow bioelectrochemical reactor equipped with an anion exchange membrane (AEM) successfully overcomes the methanogens reactivation, allowing for a continuous acetate production. The AEM membrane guaranteed the migration of the acetate from the biological compartment and its concentration in the abiotic chamber avoiding its consumption by acetoclastic methanogenesis. The system allowed an acetate concentration of 1745 ± 30 mg/L in the abiotic chamber, nearly five times the concentration measured in the cathodic chamber.

Sustainable technologies for energy production and storage are currently in great demand. Bioelectrochemical systems (BESs) offer promising solutions for both. Several attempts have been made to improve carbon felt electrode characteristics with various pretreatments in order to enhance performance. This study was motivated by gaps in current knowledge of the impact of pretreatments on the enrichment and microbial composition of bioelectrochemical systems. Therefore, electrodes were treated with poly(neutral red), chitosan, or isopropanol in a first step and then fixed in microbial electrolysis cells (MECs). Four MECs consisting of organic substance-degrading bioanodes and methane-producing biocathodes were set up and operated in batch mode by controlling the bioanode at 400 mV vs. Ag/AgCl (3M NaCl). After 1 month of operation, Enterococcus species were dominant microorganisms attached to all bioanodes and independent of electrode pretreatment. However, electrode pretreatments led to a decrease in microbial diversity and the enrichment of specific electroactive genera, according to the type of modification used. The MEC containing isopropanol-treated electrodes achieved the highest performance due to presence of both Enterococcus and Geobacter. The obtained results might help to select suitable electrode pretreatments and support growth conditions for desired electroactive microorganisms, whereby performance of BESs and related applications, such as BES-based biosensors, could be enhanced.

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.

Electro-fermentation (EF) is an upcoming technology that can control the metabolism of exoelectrogenic bacteria (i.e., bacteria that transfer electrons using an extracellular mechanism). The fermenter consists of electrodes that act as sink and source for the production and movement of electrons and protons, thus generating electricity and producing valuable products. The conventional process of fermentation has several drawbacks that restrict their application and economic viability. Additionally, metabolic reactions taking place in traditional fermenters are often redox imbalanced. Almost all metabolic pathways and microbial strains have been studied, and EF can electrochemically control this. The process of EF can be used to optimize metabolic processes taking place in the fermenter by controlling the redox and pH imbalances and by stimulating carbon chain elongation or breakdown to improve the overall biomass yield and support the production of a specific product. This review briefly discusses microbe-electrode interactions, electro-fermenter designs, mixed-culture EF, and pure culture EF in industrial applications, electro methanogenesis, and the various products that could be hence generated using this process.

Constructed wetland-microbial electrochemical snorkel (CW-MES) systems, which are short-circuited microbial fuel cells (MFC), have emerged as a novel tool for wastewater management, although the system mechanisms are insufficiently studied in process-based or environmental contexts. Based on quantitative polymerase chain reaction assays, we assessed the prevalence of different nitrogen removal processes for treating nitrate-rich waters with varying cathode materials (stainless steel, graphite felt, and copper) and sizes in the CW-MES systems and correlated them to the changes of N2O emissions. The nitrate and nitrite removal efficiencies were in range of 40% to 75% and over 98%, respectively. In response to the electrochemical manipulation, the abundances of most of the nitrogen-transforming microbial groups decreased in general. Graphite felt cathodes supported nitrifiers, but nirK-type denitrifiers were inhibited. Anaerobic ammonium oxidation (ANAMMOX) bacteria were less abundant in the electrochemically manipulated treatments compared to the controls. ANAMMOX and denitrification are the main nitrogen reducers in CW-MES systems. The treatments with 1:1 graphite felt, copper, plastic, and stainless-steel cathodes showed higher N2O emissions. nirS- and nosZI-type denitrifiers are mainly responsible for producing and reducing N2O emissions, respectively. Hence, electrochemical manipulation supported dissimilatory nitrate reduction to ammonium (DNRA) microbes may play a crucial role in producing N2O in CW-MES systems.

Membrane separators are key elements of microbial fuel cells (MFCs), especially of those constructed in a dual-chamber configuration. Until now, membranes made of Nafion have been applied the most widely to set-up MFCs. However, there is a broader agreement in the literature that Nafion is expensive and in many cases, does not meet the actual (mainly mass transfer-specific) requirements demanded by the process and users. Driven by these issues, there has been notable progress in the development of alternative materials for membrane fabrication, among which those relying on the deployment of ionic liquids are emerging. In this review, the background of and recent advances in ionic liquid-containing separators, particularly supported ionic liquid membranes (SILMs), designed for MFC applications are addressed and evaluated. After an assessment of the basic criteria to be fulfilled by membranes in MFCs, experiences with SILMs will be outlined, along with important aspects of transport processes. Finally, a comparison with the literature is presented to elaborate on how MFCs installed with SILM perform relative to similar systems assembled with other, e.g., Nafion, membranes.

A Plant Microbial Fuel Cell (Plant-MFCs) has been studied both in the lab and in a field. So far, field studies were limited to a more conventional Plant-MFC design, which submerges the anode in the soil and places the cathode above the soil surface. However, for a large scale application a tubular Plant-MFC is considered more practical since it needs no topsoil excavation. In this study, 1 m length tubular design Plant-MFC was installed in triplicate in a paddy field located in West Kalimantan, Indonesia. The Plant-MFC reactors were operated for four growing seasons. The rice paddy was grown in a standard cultivation process without any additional treatment due to the reactor instalation. An online data acquisition using LoRa technology was developed to investigate the performance of the tubular Plant-MFC over the final whole rice paddy growing season. Overall, the four crop seasons, the Plant-MFC installation did not show a complete detrimental negative effect on rice paddy growth. Based on continuous data analysis during the fourth crop season, a continuous electricity generation was achieved during a wet period in the crop season. Electricity generation dynamics were observed before, during and after the wet periods that were explained by paddy field management. A maximum daily average density from the triplicate Plant-MFCs reached 9.6 mW/m2 plant growth area. In one crop season, 9.5−15 Wh/m2 electricity can be continuously generated at an average of 0.4 ± 0.1 mW per meter tube. The Plant-MFC also shows a potential to be used as a bio sensor, e.g., rain event indicator, during a dry period between the crop seasons.

Industrial fermentation in aerobic processes is plagued by high costs due to gas transfer limitations and substrate oxidation to CO2. It has been a longstanding challenge to engineer an obligate aerobe organism, such as Pseudomonas putida, into an anaerobe to facilitate its industrial application. However, the progress in this field is limited, due to the poor understanding of the constraints restricting its anoxic phenotype. In this paper, we provide a methodological description of a novel cultivation technology for P. putida under anaerobic conditions, using the so-called microbial electrochemical technology within a bioelectrochemical system. By using an electrode as the terminal electron acceptor (mediated via redox chemicals), glucose catabolism could be activated without oxygen present. This (i) provides an anoxic-producing platform for sugar acid production at high yield and (ii) more importantly, enables systematic and quantitative characterizations of the phenotype of P. putida in the absence of molecular oxygen. This unique electrode-based cultivation approach offers a tool to understand and in turn engineer the anoxic phenotype of P. putida and possibly also other obligate aerobes.

The conversion of C1 gas feedstock, such as carbon monoxide (CO), to useful platform chemicals has attracted considerable interest in industrial biotechnology. One conversion method is electrode-based electron transfer to microorganisms using bioelectrochemical systems (BESs). In this BES system, acetate is the predominant component of various volatile fatty acids (VFAs). To appropriately separate and concentrate the acetate produced, a BES-type electrodialysis cell with an anion exchange membrane was constructed and evaluated under various operational conditions, such as applied external current, acetate concentration, and pH. A high acetate flux of 23.9 mmol/m2∙h was observed under a −15 mA current in an electrodialysis-based bioelectrochemical system. In addition, the initial acetate concentration affected the separation efficiency and transportation rate. The maximum flux appeared at 48.6 mmol/m2∙h when the acetate concentration was 100 mM, whereas the effects of the initial pH of the anolyte were negligible. The acetate flux was 14.9 mmol/m2∙h when actual fermentation broth from BES-based CO fermentation was used as a catholyte. A comparison of the synthetic broth with the actual fermentation broth suggests that unknown substances and metabolites produced from the previous bioconversion process interfere with electrodialysis. These results provide information on the optimal conditions for the separation of VFAs produced by C1 gas fermentation through electrodialysis and a combination of a BES and electrodialysis.

Background: Bioelectrochemical systems (BESs) are emerging energy-effective and environment-friendly technologies. Different applications of BESs are able to effectively minimize wastes and treat wastewater while simultaneously recovering electricity, biohydrogen and other value-added chemicals via specific redox reactions. Although there are many studies that have greatly advanced the performance of BESs over the last decade, research and reviews on agriculture-relevant applications of BESs are very limited. Considering the increasing demand for food, energy and water due to human population expansion, novel technologies are urgently needed to promote productivity and sustainability in agriculture. Methodology: This review study is based on an extensive literature search regarding agriculture-related BES studies mainly in the last decades (i.e., 2009⁻2018). The databases used in this review study include Scopus, Google Scholar and Web of Science. The current and future applications of bioelectrochemical technologies in agriculture have been discussed. Findings/Conclusions: BESs have the potential to recover considerable amounts of electric power and energy chemicals from agricultural wastes and wastewater. The recovered energy can be used to reduce the energy input into agricultural systems. Other resources and value-added chemicals such as biofuels, plant nutrients and irrigation water can also be produced in BESs. In addition, BESs may replace unsustainable batteries to power remote sensors or be designed as biosensors for agricultural monitoring. The possible applications to produce food without sunlight and remediate contaminated soils using BESs have also been discussed. At the same time, agricultural wastes can also be processed into construction materials or biochar electrodes/electrocatalysts for reducing the high costs of current BESs. Future studies should evaluate the long-term performance and stability of on-farm BES applications.

Microbial electrochemical technologies provide sustainable wastewater treatment and energy production. Despite significant improvements in the power output of microbial fuel cells (MFCs), this technology is still far from practical applications. Extracting electrical energy and harvesting valuable products by electroactive bacteria (EAB) in bioelectrochemical systems (BESs) has emerged as an innovative approach to address energy and environmental challenges. Thus, maximizing power output and resource recovery is highly desirable for sustainable systems. Insights into the electrode-microbe interactions may help to optimize the performance of BESs for envisioned applications, and further validation by bioelectrochemical techniques is a prerequisite to completely understand the electro-microbiology. This review summarizes various extracellular electron transfer mechanisms involved in BESs. The significant role of characterization techniques in the advancement of the electro-microbiology field is discussed. Finally, diverse applications of BESs, such as resource recovery, and contributions to the pursuit of a more sustainable society are also highlighted.

Biochemical oxygen demand (BOD) is a widely used index of water quality in wastewater treatment; however, conventional measurement methods are time-consuming. In this study, we analyzed a novel flame-oxidized stainless steel anode (FO-SSA) for use as the probe of bioelectrochemical system (BES)-based biosensors to monitor the BOD of treated swine wastewater. A thinner biofilm formed on the FO-SSA compared with that on a common carbon-cloth anode (CCA). The FO-SSA was superior to the CCA in terms of rapid sensing; the response time of the FO-SSA to obtain the value of R2 > 0.8 was 1 h, whereas the CCA required 4 h. These results indicate that the FO-SSA offers better performance than traditional CCAs in BES biosensors and can be used to improve biomonitoring of wastewater.

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.

Industrial fermentation in aerobic processes is plagued by high costs due to gas transfer limitations and substrate oxidation to CO 2 . It has been a longstanding challenge to engineer an obligate aerobe organism, such as Pseudomonas putida , into an anaerobe to facilitate its industrial application. However, the progress in this field is limited, due to the poor understanding of the constraints restricting its anoxic phenotype. In this paper, we provide a methodological description of a novel cultivation technology for P. putida under anaerobic conditions, using the so-called microbial electrochemical technology within a bioelectrochemical system. By using an electrode as the terminal electron acceptor (mediated via redox chemicals), glucose catabolism could be activated without oxygen present. This (i) provides an anoxic-producing platform for sugar acid production at high yield and (ii) more importantly, enables systematic and quantitative characterizations of the phenotype of P. putida in the absence of molecular oxygen. This unique electrode-based cultivation approach offers a tool to understand and in turn engineer the anoxic phenotype of P. putida and possibly also other obligate aerobes.