The periodic table of photosynthetic purple non-sulfur bacteria: intact cell-metal ions interactions.

Photosynthetic purple non-sulfur bacteria (PNB) have been widely utilized as model organisms to study bacterial photosynthesis. More recently, the remarkable resistance of these microorganisms to several metals ions called particular interest. As a result, several research efforts were directed toward clarifying the interactions of metal ions with PNB. The mechanisms of metal ions active uptake and bioabsorption have been studied in detail, unveiling that PNB enable harvesting and removing various toxic ions, thus fostering applications in environmental remediation. Herein, we present the most important achievements in the understanding of intact cell-metal ions interactions and the approaches utilized to study such processes. Following, the application of PNB-metal ions interactions toward metal removal from contaminated environments is presented. Finally, the possible coupling of PNB with abiotic electrodes to obtain biohybrid electrochemical systems is proposed as a sustainable pathway to tune and enhance metal removal and monitoring.

Fundamentals, Applications, and Future Directions of Bioelectrocatalysis.

Bioelectrocatalysis is an interdisciplinary research field combining biocatalysis and electrocatalysis via the utilization of materials derived from biological systems as catalysts to catalyze the redox reactions occurring at an electrode. Bioelectrocatalysis synergistically couples the merits of both biocatalysis and electrocatalysis. The advantages of biocatalysis include high activity, high selectivity, wide substrate scope, and mild reaction conditions. The advantages of electrocatalysis include the possible utilization of renewable electricity as an electron source and high energy conversion efficiency. These properties are integrated to achieve selective biosensing, efficient energy conversion, and the production of diverse products. This review seeks to systematically and comprehensively detail the fundamentals, analyze the existing problems, summarize the development status and applications, and look toward the future development directions of bioelectrocatalysis. First, the structure, function, and modification of bioelectrocatalysts are discussed. Second, the essentials of bioelectrocatalytic systems, including electron transfer mechanisms, electrode materials, and reaction medium, are described. Third, the application of bioelectrocatalysis in the fields of biosensors, fuel cells, solar cells, catalytic mechanism studies, and bioelectrosyntheses of high-value chemicals are systematically summarized. Finally, future developments and a perspective on bioelectrocatalysis are suggested.

Unbranched Hybrid Conducting Redox Polymers for Intact Chloroplast-Based Photobioelectrocatalysis.

Photobioelectrocatalysis (PBEC) adopts the sophistication and sustainability of photosynthetic units to convert solar energy into electrical energy. However, the electrically insulating outer membranes of photosynthetic units hinder efficient extracellular electron transfer from photosynthetic redox centers to an electrode in photobioelectrocatalytic systems. Among the artificial redox-mediating approaches used to enhance electrochemical communication at this biohybrid interface, conducting redox polymers (CRPs) are characterized by high intrinsic electric conductivities for efficient charge transfer. A majority of these CRPs constitute peripheral redox pendants attached to a conducting backbone by a linker. The consequently branched CRPs necessitate maintaining synergistic interactions between the pendant, linker, and backbone for optimal mediator performance. Herein, an unbranched, metal-free CRP, polydihydroxy aniline (PDHA), which has its redox moiety embedded in the polymer mainchain, is used as an exogenous redox mediator and an immobilization matrix at the biohybrid interface. As a proof of concept, the relatively complex membrane system of spinach chloroplasts is used as the photobioelectrocatalyst of choice. A "mixed" deposition of chloroplasts and PDHA generated a 2.4-fold photocurrent density increment. An alternative "layered" PDHA-chloroplast deposition, which was used to control panchromatic light absorbance by the intensely colored PDHA competing with the photoactivity of chloroplasts, generated a 4.2-fold photocurrent density increment. The highest photocurrent density recorded with intact chloroplasts was achieved by the "layered" deposition when used in conjunction with the diffusible redox mediator 2,6-dichlorobenzoquinone (-48 ± 3 µA cm-2). Our study effectively expands the scope of germane CRPs in PBEC, emphasizing the significance of the rational selection of CRPs for electrically insulating photobioelectrocatalysts and of the holistic modulation of the CRP-mediated biohybrids for optimal performance.

Rational design of artificial redox-mediating systems toward upgrading photobioelectrocatalysis.

Photobioelectrocatalysis has recently attracted particular research interest owing to the possibility to achieve sunlight-driven biosynthesis, biosensing, power generation, and other niche applications. However, physiological incompatibilities between biohybrid components lead to poor electrical contact at the biotic-biotic and biotic-abiotic interfaces. Establishing an electrochemical communication between these different interfaces, particularly the biocatalyst-electrode interface, is critical for the performance of the photobioelectrocatalytic system. While different artificial redox mediating approaches spanning across interdisciplinary research fields have been developed in order to electrically wire biohybrid components during bioelectrocatalysis, a systematic understanding on physicochemical modulation of artificial redox mediators is further required. Herein, we review and discuss the use of diffusible redox mediators and redox polymer-based approaches in artificial redox-mediating systems, with a focus on photobioelectrocatalysis. The future possibilities of artificial redox mediator system designs are also discussed within the purview of present needs and existing research breadth.

Purple bacteria photo-bioelectrochemistry: enthralling challenges and opportunities.

Purple non-sulfur bacteria are anoxygenic photosynthetic microorganisms characterized by an extremely versatile metabolism, allowing them to grow in a broad variety of conditions as well as in the presence of different contaminants. This characteristic motivates the interest in their employment in photo-bioelectrochemical systems applicable in environments with dynamic physico-chemical properties. While the photochemistry of purple bacteria has been intensively studied, their photo-bioelectrochemistry and extracellular electron transfer process with an electrode surface remain largely unexplored. Herein, the process of harvesting electrons from intact purple bacteria is reviewed, and the perspective of enthralling future research possibilities is presented, placing emphasis on the major challenges in the photo-bioelectrochemistry of purple bacteria.

Online self-powered Cr(VI) monitoring with autochthonous Pseudomonas and a bio-inspired redox polymer.

The assessment of water quality is critical to implement preventive and emergency interventions aimed to limit/avoid environmental contamination and human exposure to toxic compounds. While established high-resolution techniques allow quantitative and qualitative determination of contaminants, their widespread application is not feasible due to cost, time, and need for trained personnel. In this context, the development of easy-to-implement approaches for preliminary detection of contaminants is of the utmost importance. Herein, a portable self-powered microbial electrochemical sensor enabling online monitoring of Cr(VI) is reported. The biosensor employs a bio-inspired redox mediating system to allow extracellular electron transfer between a bacterial isolate from chromium-contaminated environments and the electrode, providing a clear response to Cr(VI) presence. The biosensor shows good linearity (R2 = 0.983) and a limit of detection of 2.4 mg L-1 Cr(VI), with a sensitivity of 0.31 ± 0.02 µA cm-2 mgCr(VI)-1 L. The presented microbial bioanode architecture enhanced biosensor performance thanks to the improved "electrical wiring" between biological entities and the abiotic electrode surface. This approach could be easily implemented in engineered electrode surfaces, such as paper-based multi-anodes that maximize bacterial colonization, further improving biosensor response. Graphical abstract.

Unveiling salinity effects on photo-bioelectrocatalysis through combination of bioinformatics and electrochemistry.

Little is known about the adaptation strategies utilized by photosynthetic microorganisms to cope with salinity changes happening in the environment, and the effects on microbial electrochemical technologies. Herein, bioinformatics analysis revealed a metabolism shift in Rhodobacter capsulatus resulting from salt stress, with changes in gene expression allowing accumulation of compatible solutes to balance osmotic pressure, together with the up-regulation of the nitrogen fixation cycle, an electron sink of the photosynthetic electron transfer chain. Using the transcriptome evidence of hindered electron transfer in the photosynthetic electron transport chain induced by adaption to salinity, increased understanding of photo-bioelectrocatalysis under salt stress is achieved. Accumulation of glycine-betaine allows immediate tuning of salinity tolerance but does not provide cell stabilization, with a 40 ± 20% loss of photo-bioelectrocatalysis in a 60 min time scale. Conversely, exposure to or inducing the expression of the Rhodobacter capsulatus gene transfer agent tunes salinity tolerance and increases cell stability. This work provides a proof of concept for the combination of bioinformatics and electrochemical tools to investigate microbial electrochemical systems, opening exciting future research opportunities.

Tuning purple bacteria salt-tolerance for photobioelectrochemical systems in saline environments.

The development of photobioelectrochemical systems is an exciting field requiring a combination of electrochemical, biological and material science knowledge. One of the main advantages of applying anoxygenic photosynthetic microorganisms versus non-photosynthetic bacteria is the possibility to utilize sunlight as the energy source, while removing organic contaminants from a solution. Since bacterial cells utilize energy to maintain the intracellular osmolarity, bacterial species that do not rely on organic species as an energy source have an advantage over species requiring them for their sustainment. Herein, we discuss the possible use of Rhodobacter capsulatus, an extremely versatile photosynthetic purple bacteria, for application in environments within a range of low to moderately high salinity (0-25 g L-1 NaCl). Bacterial cells' capability to adapt to changing salinity, and effects on bioelectrochemical performance will be presented, as well as major drawbacks and research needs to drive future efforts and discussions.

Alginate-Encapsulated Bacteria for the Treatment of Hypersaline Solutions in Microbial Fuel Cells.

A microbial fuel cell (MFC) based on a new wild-type strain of Salinivibrio sp. allowed the self-sustained treatment of hypersaline solutions (100 g L-1 , 1.71 m NaCl), reaching a removal of (87±11) % of the initial chemical oxygen demand after five days of operation, being the highest value achieved for hypersaline MFC. The degradation process and the evolution of the open circuit potential of the MFCs were correlated, opening the possibility for online monitoring of the treatment. The use of alginate capsules to trap bacterial cells, increasing cell density and stability, resulted in an eightfold higher power output, together with a more stable system, allowing operation up to five months with no maintenance required. The reported results are of critical importance to efforts to develop a sustainable and cost-effective system that treats hypersaline waste streams and reduces the quantity of polluting compounds released.

Living Diatom Microalgae for Desiccation-Resistant Electrodes in Biophotovoltaic Devices.

Strategies of renewable energy production from photosynthetic microorganisms are gaining great scientific interest as ecosustainable alternatives to fossil fuel depletion. Green microalgae have been thoroughly investigated as living components to convert solar energy into photocurrent in biophotovoltaic (BPV) cells. Conversely, the suitability of diatoms in BPV cells has been almost completely unexplored so far, despite being the most abundant class of photosynthetic microorganisms in phytoplankton and of their good adaptability and resistance to harsh environmental conditions, including dehydration, high salinity, nutrient starvation, temperature, or pH changes. Here, we demonstrate the suitability of a series of diatom species (Phaeodactylum tricornutum, Thalassiosira weissflogii, Fistulifera pelliculosa, and Cylindrotheca closterium), to act as biophotoconverters, coating the surface of indium tin oxide photoanodes in a model BPV cell. Effects of light intensity, cell density, total chlorophyll content, and concentration of the electrochemical mediator on photocurrent generation efficiency were investigated. Noteworthily, biophotoanodes coated with T. weissflogii diatoms are still photoactive after 15 days of dehydration and four rewetting cycles, contrary to analogue electrodes coated with the model green microalga Dunaliella tertiolecta. These results provide the first evidence that diatoms are suitable photosynthetic microorganisms for building highly desiccation-resistant biophotoanodes for durable BPV devices.

Polydopamine-immobilized yeast cells for portable electrochemical biosensors applied in environmental copper sensing.

The coupling of biological organisms with electrodes enables the development of sustainable, low cost, and potentially self-sustained biosensors. A critical aspect is to obtain portable bioelectrodes where the biological material is immobilized on the electrode surface to be utilized on demand. Herein, we developed an approach for the rapid entrapment and immobilization of metabolically active yeast cells in a biocompatible polydopamine layer, which does not require a separate and time-consuming synthesis. The reported approach allows obtaining the "electrical wire" of intact and active yeast cells with resulting current generation from glucose oxidation. Additionally, the electrochemical performance of the biohybrid yeast-based system has been characterized in the presence of CuSO4, a widely used pesticide, in the environmentally relevant concentration range of 20-100 µM. The system enabled the rapid preliminary monitoring of the contaminant based on variations in current generation, with a limit of detection of 12.5 µM CuSO4. The present approach for the facile preparation of portable yeast-based electrochemical biosensors paves the way for the future development of sustainable systems for environmental monitoring.

Enzymatic and Microbial Electrochemistry: Approaches and Methods.

The coupling of enzymes and/or intact bacteria with electrodes has been vastly investigated due to the wide range of existing applications. These span from biomedical and biosensing to energy production purposes and bioelectrosynthesis, whether for theoretical research or pure applied industrial processes. Both enzymes and bacteria offer a potential biotechnological alternative to noble/rare metal-dependent catalytic processes. However, when developing these biohybrid electrochemical systems, it is of the utmost importance to investigate how the approaches utilized to couple biocatalysts and electrodes influence the resulting bioelectrocatalytic response. Accordingly, this tutorial review starts by recalling some basic principles and applications of bioelectrochemistry, presenting the electrode and/or biocatalyst modifications that facilitate the interaction between the biotic and abiotic components of bioelectrochemical systems. Focus is then directed toward the methods used to evaluate the effectiveness of enzyme/bacteria-electrode interaction and the insights that they provide. The basic concepts of electrochemical methods widely employed in enzymatic and microbial electrochemistry, such as amperometry and voltammetry, are initially presented to later focus on various complementary methods such as spectroelectrochemistry, fluorescence spectroscopy and microscopy, and surface analytical/characterization techniques such as quartz crystal microbalance and atomic force microscopy. The tutorial review is thus aimed at students and graduate students approaching the field of enzymatic and microbial electrochemistry, while also providing a critical and up-to-date reference for senior researchers working in the field.

Bio-Inspired Redox-Adhesive Polydopamine Matrix for Intact Bacteria Biohybrid Photoanodes.

Interfacing intact and metabolically active photosynthetic bacteria with abiotic electrodes requires both establishing extracellular electron transfer and immobilizing the biocatalyst on electrode surfaces. Artificial approaches for photoinduced electron harvesting through redox polymers reported in literature require the separate synthesis of artificial polymeric matrices and their subsequent combination with bacterial cells, making the development of biophotoanodes complex and less sustainable. Herein, we report a one-pot biocompatible and sustainable approach, inspired by the byssus of mussels, that provides bacterial cells adhesion on multiple surfaces under wet conditions to obtain biohybrid photoanodes with facilitated photoinduced electron harvesting. Purple bacteria were utilized as a model organism, as they are of great interest for the development of photobioelectrochemical systems for H2 and NH3 synthesis, biosensing, and bioremediation purposes. The polydopamine matrix preparation strategy allowed the entrapment of active purple bacteria cells by initial oxygenic polymerization followed by electrochemical polymerization. Our results unveil that the deposition of bacterial cells with simultaneous polymerization of polydopamine on the electrode surface enables a 5-fold enhancement in extracellular electron transfer at the biotic/abiotic interface while maintaining the viability of the cells. The presented approach paves the way for a more sustainable development of biohybrid photoelectrodes.

Draft Genome Sequence of Salinivibrio sp. Strain EAGSL, a Biotechnologically Relevant Halophilic Microorganism.

The halophilic bacterium Salinivibrio sp. strain EAGSL was isolated from the Great Salt Lake (Utah) for use in microbial electrochemical technologies experiencing fluctuating salt concentrations. Genome sequencing was performed with Ion Torrent technology, and the assembled genome reported here is 3,234,770 bp with a GC content of 49.41%.

Chloroplast biosolar cell and self-powered herbicide monitoring.

Utilizing chloroplasts in biosolar cells offers a sustainable approach for sunlight harvesting. However, the limited electrochemical communication between these biological entities and an electrode surface has led to complex device setups, hindering their application in the field. Herein, a cross-linker enables a simple photoanode architecture with enhanced photoexcited electron transfer between chloroplasts and abiotic electrodes. The improved "wiring" of the photosynthetic electron transfer chain resulted in a five-fold increase in the biophotocurrent. The biophotoanode is applied in a Pt-free, portable biosolar cell allowing the in situ self-powered monitoring of diuron within limits set by the Environmental Protection Agency.

Purple Bacteria and 3D Redox Hydrogels for Bioinspired Photo-bioelectrocatalysis.

A major challenge for the implementation of intact bacterial cells in photo-bioelectrochemical systems remains the hindered extracellular electron transfer. This study focuses on purple bacteria, photosynthetic microorganisms particularly interesting for the development of bioelectrochemical systems because of their versatile metabolisms. Although soluble monomeric redox mediators have been proven as effective systems for electron transfer mediation, their application in the field is not preferable owing to their toxicity and unwanted release into the environment. An abiotic/biotic photoanode is reported in which a bioinspired redox mediating system is implemented in a 3D geometry allowing to "electrically wire" intact bacterial cells. The 3D photoanode decreased the overpotential required for harvesting photoexcited electrons, operating at +0.073 V versus the saturated calomel electrode (SCE). Accordingly, the overpotential was significantly reduced compared with a pioneering Os-redox polymer reported in literature, which required operation at +0.303 V versus SCE. These results provide the basis for further development of bio-photoanodes for light-powered biosensing and power generation.

Advancing the fundamental understanding and practical applications of photo-bioelectrocatalysis.

Photo-bioelectrocatalysis combines the natural and highly sophisticated process of photosynthesis in biological entities with an abiotic electrode surface, to perform semi-artificial photosynthesis. However, challenges must be overcome, from the establishment and understanding of the photoexcited electron harvesting process at the electrode to the electrochemical characterization of these biotic/abiotic systems, and their subsequent tuning for enhancing energy generation (chemical and/or electrical). This Feature Article discusses the various approaches utilized to tackle these challenges, particularly focusing on powerful multi-disciplinary approaches for understanding and improving photo-bioelectrocatalysis. Among them is the combination of experimental evidence and quantum mechanical calculations, the use of bioinformatics to understand photo-bioelectrocatalysis at a metabolic level, or bioengineering to improve and facilitate photo-bioelectrocatalysis. Key aspects for the future development of photo-bioelectrocatalysis are presented alongside future research needs and promising applications of semi-artificial photosynthesis.

Microbial fuel cells in saline and hypersaline environments: Advancements, challenges and future perspectives.

This review is aimed to report the possibility to utilize microbial fuel cells for the treatment of saline and hypersaline solutions. An introduction to the issues related with the biological treatment of saline and hypersaline wastewater is reported, discussing the limitation that characterizes classical aerobic and anaerobic digestions. The microbial fuel cell (MFC) technology, and the possibility to be applied in the presence of high salinity, is discussed before reviewing the most recent advancements in the development of MFCs operating in saline and hypersaline conditions, with their different and interesting applications. Specifically, the research performed in the last 5years will be the main focus of this review. Finally, the future perspectives for this technology, together with the most urgent research needs, are presented.

Self-Powered Biosensors.

Self-powered electrochemical biosensors utilize biofuel cells as a simultaneous power source and biosensor, which simplifies the biosensor system, because it no longer requires a potentiostat, power for the potentiostat, and/or power for the signaling device. This review article is focused on detailing the advances in the field of self-powered biosensors and discussing their advantages and limitations compared to other types of electrochemical biosensors. The review will discuss self-powered biosensors formed from enzymatic biofuel cells, organelle-based biofuel cells, and microbial fuel cells. It also discusses the different mechanisms of sensing, including utilizing the analyte being the substrate/fuel for the biocatalyst, the analyte binding the biocatalyst to the electrode surface, the analyte being an inhibitor of the biocatalyst, the analyte resulting in the blocking of the bioelectrocatalytic response, the analyte reactivating the biocatalyst, Boolean logic gates, and combining affinity-based biorecognition elements with bioelectrocatalytic power generation. The final section of this review details areas of future investigation that are needed in the field, as well as problems that still need to be addressed by the field.

Lag Time Spectrophotometric Assay for Studying Transport Limitation in Immobilized Enzymes.

Enzymes are promising catalysts for bioprocessing. For instance, the enzymatic capture of CO2 using carbonic anhydrase (CA) is a carbon capture approach that allows obtaining bicarbonate (HCO3 -) with no high-energy input required. However, application in a commercially viable biotechnology requires sufficient enzymatic lifetime. Although enzyme stabilization can be achieved by different immobilization techniques, most of them are not commercially viable because of transport limitations induced by the immobilization method. Therefore, it is necessary to develop assays for evaluating the role of immobilization on transport limitations. Herein, we describe the development of a fast and reproducible assay for screening immobilized CA by means of absorbance measurement using a computer-controlled microplate reader in stop-flow format. The automated assay allowed minimizing the required volume for analysis to 120 µL. We validated the assay by determining lag times and activities for three immobilization techniques (modified Nafion, hydrogels, and enzyme precipitates), of which linear polyethyleneimine hydrogel showed outstanding performance for CA immobilization.