Acetogenesis to ethanologenesis: facilitating NADH oxidation via reductive acetate uptake.

(Homo)acetogens, including Clostridium spp., represent an enigma in metabolic flexibility and diversity. Eubacterium callanderi KIST612 is an acetogen that produces n-butyrate with carbon monoxide (CO) as the carbon and energy source; however, the production route is unknown.

A Redox-Enzyme Integrated Microbial Fuel Cell Design Using the Surface Display System in MR-1.

A biofuel cell is an electrochemical device using exoelectrogen or biocatalysts to transfer electrons from redox reactions to the electrodes. While wild-type microbes and natural enzymes are often employed as exoelectrogen and biocatalysts, genetically engineered or modified organisms have been developed to enhance exoelectrogen activity. Here, we demonstrated a redox-enzyme integrated microbial fuel cell (REI-MFC) design based on an exoelectrogen-enhancing strategy that reinforces the electrogenic activity of MR1 by displaying an extra redox enzyme on the cell surface.

Advanced strategies for enzyme-electrode interfacing in bioelectrocatalytic systems.

Advances in protein engineering-enabled enzyme immobilization technologies have significantly improved enzyme-electrode wiring in enzymatic electrochemical systems, which harness natural biological machinery to either generate electricity or synthesize biochemicals. In this review, we provide guidelines for designing enzyme-electrodes, focusing on how performance variables change depending on electron transfer (ET) mechanisms. Recent advancements in enzyme immobilization technologies are summarized, highlighting their contributions to extending enzyme-electrode sustainability (up to months), enhancing biosensor sensitivity, improving biofuel cell performance, and setting a new benchmark for turnover frequency in bioelectrocatalysis.

NADH-dependent CO reductase on graphite for capacitive electrocatalytic interfacing mediated by solid-binding peptide.

NAD/NADH-dependent CO reductase (CR) adapted from Candida methylica (E.C. 1.

Prevalence of Escherichia coli in electrogenic biofilm on activated carbon in microbial fuel cell.

For a better understanding of the distribution of depth-dependent electrochemically active bacteria at in the anode zone, a customized system in a microbial fuel cell (MFC) packed with granular activated carbon (GAC) was developed and subsequently optimized via electrochemical tests. The constructed MFC system was sequentially operated using two types of matrice solutions: artificially controlled compositions (i.e.

Orientation-Controllable Enzyme Cascade on Electrode for Bioelectrocatalytic Chain Reaction.

The accomplishment of concurrent interenzyme chain reaction and direct electric communication in a multienzyme-electrode is challenging since the required condition of multienzymatic binding conformation is quite complex. In this study, an enzyme cascade-induced bioelectrocatalytic system has been constructed using solid binding peptide (SBP) as a molecular binder that coimmobilizes the invertase (INV) and flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase gamma-alpha complex (GDHγα) cascade system on a single electrode surface. The SBP-fused enzyme cascade was strategically designed to induce diverse relative orientations of coupling enzymes while enabling efficient direct electron transfer (DET) at the FAD cofactor of GDHγα and the electrode interface.

Regulatory transcription factor (CooA)-driven carbon monoxide partial pressure sensing whole-cell biosensor.

We designed and constructed a whole-cell biosensor capable of detecting the presence and quantity of carbon monoxide (CO) using the CO regulatory transcription factor. This biosensor utilizes CooA, a CO-sensing transcription regulator that activates the expression of carbon monoxide dehydrogenase (CODH), to detect the presence of CO and respond by triggering the expression of a GUS reporter protein (β-glucuronidase). The GUS reporter protein is expressed from a CO-induced CooA-binding promoter (P) by CooA and enables the effective colorimetric detection of CO.

Acetogen and acetogenesis for biological syngas valorization.

The bioconversion of syngas using (homo)acetogens as biocatalysts shows promise as a viable option due to its higher selectivity and milder reaction conditions compared to thermochemical conversion. The current bioconversion process operates primarily to produce C2 chemicals (e.g.

Genome-Based Reclassification of Strain KIST612, Previously Classified as , into a New Strain of .

The strain KIST612, initially identified as , was a suspected member of due to differences in phenotype, genotype, and average nucleotide identity (ANI). Here, we found that ATCC 8486 and KIST612 are genetically different in their central metabolic pathways, such as that of carbon metabolism. Although 16S rDNA sequencing of KIST612 revealed high identity with ATCC 8486 (99.

Task-specific polymeric membranes to achieve high gas-liquid mass transfer.

In the current study, Polyimide (P84)-based polymeric membranes were fabricated and used as spargers in the bubble column reactor (BCR) to get a high gas-liquid mass transfer (GL-MT) rate of oxygen in water. Different polymeric membranes were fabricated by incorporating polyvinyl pyrrolidone (PVP) as a porogen and a Zeolitic Imidazolate Framework (ZIF-8) to induce high porosity and hydrophobicity in the membranes. The GL-MT efficiency of membranes was evaluated by measuring the overall volumetric mass transfer coefficient (ka) of oxygen in air.

Protocol for construction and characterization of direct electron transfer-based enzyme-electrode using gold binding peptide as molecular binder.

Here, we present a protocol for constructing direct electron transfer (DET)-based enzyme-electrodes using gold-binding peptide (GBP). We describe fusion of four GBPs to flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase gamma-alpha complex (GDHγα), as model oxidoreductase, to generate four GDHγα variants. We then detail the measurements of catalytic and bioelectrochemical properties of these GDHγα variants on electrode together with surface morphology of GDHγα variants immobilized on gold surface.

Control of carbon monoxide dehydrogenase orientation by site-specific immobilization enables direct electrical contact between enzyme cofactor and solid surface.

Controlling the orientation of redox enzymes on electrode surfaces is essential in the development of direct electron transfer (DET)-based bioelectrocatalytic systems. The electron transfer (ET) distance varies according to the enzyme orientation when immobilized on an electrode surface, which influences the interfacial ET rate. We report control of the orientation of carbon monoxide dehydrogenase (CODH) as a model enzyme through the fusion of gold-binding peptide (gbp) at either the N- or the C-terminus, and at both termini to strengthen the binding interactions between the fusion enzyme and the gold surface.

Peptide sequence-driven direct electron transfer properties and binding behaviors of gold-binding peptide-fused glucose dehydrogenase on electrode.

Oriented enzyme immobilization on electrodes is crucial for interfacial electrical coupling of direct electron transfer (DET)-based enzyme-electrode systems. As inorganic-binding peptides are introduced as molecular binders and enzyme-orienting agents, inorganic-binding peptide-fused enzymes should be designed and constructed to achieve efficient DET. In this study, it is aimed to compare the effects of various gold-binding peptides (GBPs) fused to enzymes on electrocatalytic activity, bioactivity, and material-binding behaviors.

Metabolism perturbation Causedby the overexpression of carbon monoxide dehydrogenase/Acetyl-CoA synthase gene complex accelerated gas to acetate conversion rate ofEubacterium limosumKIST612.

Microbial conversion of carbon monoxide (CO) to acetate is a promising upcycling strategy for carbon sequestration. Herein, we demonstrate that CO conversion and acetate production rates of Eubacterium limosum KIST612 strain can be improved by in silico prediction and in vivo assessment. The mimicked CO metabolic model of KIST612 predicted that overexpressing the CO dehydrogenase (CODH) increases CO conversion and acetate production rates.

Functional Expression of a Mo-Cu-Dependent Carbon Monoxide Dehydrogenase (CODH) and Its Use as a Dissolved CO Bio-microsensor.

Herein, we report the heterologous expression in of a Mo-Cu-containing carbon monoxide dehydrogenase (Mo-Cu CODH) from , which resulted in an active protein catalyzing CO oxidation to CO. By supplying the growth medium with NaMoO (Mo) and CuSO (Cu), the Mo-Cu CODH metal cofactors precursors, the expressed L-subunit was found to have CO-oxidation activity even without the M- and S- subunits. This successful expression of CO-oxidizing-capable single L-subunit provides direct evidence of its role as the catalytic center of Mo-Cu CODH that has not been discovered and studied before.

Emerging trends in microbial fuel cell diversification-Critical analysis.

Global need for transformation from fossil-based to bio-based economy is constantly emerging for the production of low-carbon/renewable energy/products. Microbial fuel cell (MFC) catalysed by bio-electrochemical process gained significant attention initially for its unique potential to generate energy. Diversification of MFC is an emerging trend in the context of prioritising/enhancing product output while exploring the mechanism specificity of individual processes.

Methanol supply speeds up synthesis gas fermentation by methylotrophic-acetogenic bacterium, Eubacterium limosum KIST612.

This study analyzed the effect of methanol on the metabolism of syngas components (i.e., H and CO) by the syngas fermenting acetogenic strain E.

Microbial fuel cell driven mineral rich wastewater treatment process for circular economy by creating virtuous cycles.

The aim of this work is to study for concurrent harvesting bioelectricity and struvite mineral from mineral rich wastewater containing with nitrogen (N) and phosphorous (P) contents using MFCs and a chemical precipitation system. Whole reaction was constructed to sequentially run hybrid reactor (consisting of MFCs and struvite precipitation), gravitational sedimentation, nitrogen purging and MFCs. The MFCs generated around 6.

Biosensing and electrochemical properties of flavin adenine dinucleotide (FAD)-Dependent glucose dehydrogenase (GDH) fused to a gold binding peptide.

In the present work, direct electron transfer (DET) based biosensing system for the determination of glucose has been fabricated by utilizing gold binding peptide (GBP) fused flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) from Burkholderia cepacia. The GBP fused FAD-GDH was immobilized on the working electrode surface of screen-printed electrode (SPE) which consists of gold working electrode, a silver pseudo-reference electrode and a platinum counter electrode, to develop the biosensing system with compact design and favorable sensing ability. The bioelectrochemical and mechanical properties of GBP fused FAD-GDH (GDH-GBP) immobilized SPE (GDH-GBP/Au) were investigated.

Behavior of CO-water mass transfer coefficient in membrane sparger-integrated bubble column for synthesis gas fermentation.

The gas-liquid mass transfer coefficient (ka) of O was investigated in a bubble column reactor (BCR) using a sintered gas filter (SF), ceramic membrane module (CMM), and hollow fiber membrane module (HFM), which have different ranges of gas supply areas. ka was enhanced by increasing flow rate in all of the spargers. Different responses when changing the gas supply area were obtained depending on the sparger type.

Controlling Voltage Reversal in Microbial Fuel Cells.

Microbial fuel cell (MFC) systems have been developed for potential use as power sources, along with several other applications, with bacteria as the prime factor enabling electrocatalytic activity. Limited voltage and current production from unit cells limit their practical applicability, so stacking multiple MFCs has been proposed as a way to increase power production. Special attention is paid to voltage reversal (VR), a common occurrence in stacked MFCs, and to identifying the mechanisms underlying this phenomenon.

A simultaneous gas feeding and cell-recycled reaction (SGCR) system to achieve biomass boosting and high acetate titer in microbial carbon monoxide fermentation.

This study employed a simultaneous gas feeding and cell-recycled reaction (SGCR) system to ferment CO using Eubacterium limosum KIST612. A bubble column reactor was equipped with an ex-situ hollow fiber membrane module to enable cell recycling. The internal gas circulation rate was adjusted by controlling the pump speed to provide sufficient gas supplement to the microorganism.

Determination of optimum electrical connection mode for multi-electrode-embedded microbial fuel cells coupled with anaerobic digester for enhancement of swine wastewater treatment efficiency and energy recovery.

In this work, three multi-electrode-embedded microbial fuel cells (MFCs) were connected sequentially and operated in series and parallel modes, fed by effluent of an anaerobic digester continuously operated using swine wastewater. The anaerobic digester achieved ~0.75 CH L d while removing 71.