Electron Flow Pathways Research Articles

Biogenic amorphous FeOOH activated additional intracellular electron flow pathways for accelerating reductive dechlorination of tetrachloroethylene

Dissimilatory iron-reducing bacteria (DIRB) with extracellular electron transfer (EET) capabilities have shown significant potential for bioremediating halogenated hydrocarbon contaminated sites rich in iron and humic substances. However, the role and microbial molecular mechanisms of iron-humic acid (Fe-HA) complexes in the reductive dehalogenation process of DIRB remains inadequately elucidated. In this study, we developed a sustainable carbon cycling approach using Fe-HA complexes to modulate the electron flux from sawdust (SD), enabling almost complete reductive dechlorination by most DIRB (e.g., Shewanella oneidensis MR-1) that lack complex iron-sulfur molybdo enzymes. The SD-Fe-HA/MR-1 system achieved a 96.52% removal efficiency of tetrachloroethylene (PCE) at concentrations up to 250 μmol/L within 60 days. Material characterization revealed that DIRB facilitated the hydrolysis of macromolecular carbon sources by inducing the formation of amorphous ferrihydrite (FeOOH) in Fe-HA complexes. More importantly, the bioavailable FeOOH activated additional intracellular electron flow pathways, increasing the activity of potential dehalogenases. Transcriptome further highlight the innovative role of biogenic amorphous FeOOH in integrating intracellular redox metabolism with extracellular charge exchange to facilitate reductive dechlorination in DIRB. These findings provide novel insights into accelerating reductive dechlorination in-situ contaminated sites lacking obligate dehalogenating bacteria.

Instant rerouting of photosynthetic electron transport to O2 reduction after the plasma membrane excitation of Chara in the presence of methyl viologen

—Action potential (AP) of excitable plant cells is an important signaling event that can differentially alter physicochemical and physiological processes in various parts of the same cell. In giant cells of characean algae, the AP propagation has minor effect on photosynthetic electron transport in areas with high activity of plasmalemmal H+-pump but inhibits linear electron flow in regions featuring high passive H+/OH− conductance of the plasma membrane (PM). Uneven spatial distributions of local periplasmic and cytoplasmic pH facilitate the operation of distinct (CO2-dependent and O2-mediated) pathways of photoinduced electron flow, which presumably accounts for differential influence of AP on photosynthesis. The excitation of Chara australis cell in the presence of methyl viologen (MV), a redox mediator with the prooxidant action, provides a convenient model system to clarify the influence of voltage-dependent ion fluxes across PM on photosynthetic activity of chloroplasts. This study shows that permeation of MV to their target sites in chloroplasts is restricted by PM in resting cells, but MV easily passes through ionic channels opened during the PM depolarization. This gated permeation of MV gives rise to strong non-photochemical quenching, decrease in the effective quantum yield of linear electron flow, apparent O2 uptake, and, finally, the enhanced ROS production, as detected by the fluorescent probe dichlorofluorescein. Taken together, the results indicate that the AP generation in the presence of MV acts as trigger for instant redirection of photosynthetic linear electron flow from CO2-dependent route to the path of O2 reduction with the eventual formation of H2O2 as a dominant and most stable ROS form.

Insights into the azo dye decolourisation and denitrogenation in micro-electrolysis enhanced counter-diffusion biofilm system

In this study, electron transport pathways were activated and diversified by coupling counter-diffusion biofilms with micro-electrolysis for Alizarin yellow R (AYR) denitrogenation. Due to the binding of AYR to two residues of EC 4.1.3.36 with higher binding energy, the expression of EC 4.1.3.36 was down-regulated, causing the EC 3.1.2.28 and EC 2.5.1.74 for menaquinone synthesis (redox mediator) undetectable in Membrane aerated biofilm reactors (MABR). Spontaneous electron generation in the micro electrolysis-coupled MABR (ME-MABR) significantly activated two enzymes. Activated menaquinone up-regulated decolourisation related genes expression in ME-MABR, including azoR (2.12 log2), NQO1 (2.97 log2), wrbA (0.45 log2), and ndh (0.47 log2). The diversified electron flow pathways also promoted the nitrogen metabolism coding genes up-regulation, accelerating further inorganic nitrogen denitrogenation after AYR mineralisation. Compared to MABR, the decolourisation, mineralisation, and denitrogenation in ME-MABR increased by 25.80 %, 16.53 %, and 13.32 %, respectively. This study provides new insights into micro-electrolysis enhanced removal of AYR.

Lumen air pressure regulated multifunctional microbiotas in membrane-aerated biofilm reactors for simultaneous nitrogen removal and antibiotic elimination from aquaculture wastewater

In this study, two membrane-aerated biofilm reactors (MABRs) were constructed: one solely utilizing biofilm and another hybrid MABR (HMABR) incorporating both suspended-sludge and biofilm to treat low C/N aquaculture wastewater under varying lumen air pressure (LAP). Both HMABR and MABR demonstrated superior nitrogen removal than conventional aeration reactors. Reducing LAP from 10 kPa to 2 kPa could enhance denitrification processes without severely compromising nitrification, resulting in an increase in total inorganic nitrogen (TIN) removal from 50.2±3.1 % to 71.6±1.0 %. The HMABR exhibited better denitrification efficacy than MABR, underscoring its potential for advanced nitrogen removal applications. A decline in LAP led to decreased extracellular polymeric substance (EPS) production, which could potentially augment reactor performance by minimizing mass transfer resistance while maintaining microbial matrix stability and function. Gene-centric metagenomics analysis revealed decreasing LAP impacted nitrogen metabolic potentials and electron flow pathways. The enrichment of napAB at higher LAP and the presence of complete ammonia oxidation (Comammox) Nitrospira at lower LAP indicated aerobic denitrification and Comammox processes in nitrogen removal. Multifunctional microbial communities developed under LAP regulation, diversifying the mechanisms for simultaneous nitrification-denitrification. Increased denitrifying gene pool (narGHI, nirK, norB) and enzymatic activity at a low LAP can amplify denitrification by promoting denitrifying genes and electron flow towards denitrifying enzymes. Sulfamethoxazole (SMX) was simultaneously removed with efficiency up to 80.2 ± 3.7 %, mainly via biodegradation, while antibiotic resistome and mobilome were propagated. Collectively, these findings could improve our understanding of nitrogen and antibiotic removal mechanisms under LAP regulation, offering valuable insights for the effective design and operation of MABR systems in aquaculture wastewater treatment.

PGR5-LIKE PHOTOSYNTHETIC PHENOTYPE1A redox states alleviate photoinhibition during changes in light intensity.

Plants have evolved photosynthetic regulatory mechanisms to maintain homeostasis in response to light changes during diurnal transitions and those caused by passing clouds or by wind. One such adaptation directs photosynthetic electron flow to a cyclic pathway to alleviate excess energy surges. Here, we assign a function to regulatory cysteines of PGR5-like protein 1A (PGRL1A), a constituent of the PROTON GRADIENT REGULATION5 (PGR5)-dependent cyclic electron flow (CEF) pathway. During step increases from darkness to low light intensity in Arabidopsis (Arabidopsis thaliana), the intermolecular disulfide of the PGRL1A 59-kDa complex was reduced transiently within seconds to the 28-kDa form. In contrast, step increases from darkness to high light stimulated a stable, partially reduced redox state in PGRL1A. Mutations of 2 cysteines in PGRL1A, Cys82 and Cys183, resulted in a constitutively pseudo-reduced state. The mutant displayed higher proton motive force (PMF) and nonphotochemical quenching (NPQ) than the wild type (WT) and showed altered donor and acceptor dynamic flow around PSI. These changes were found to correspond with the redox state of PGRL1A. Continuous light regimes did not affect mutant growth compared to the WT. However, under fluctuating regimes of high light, the mutant showed better growth than the WT. In contrast, in fluctuating regimes of low light, the mutant displayed a growth penalty that can be attributed to constant stimulation of CEF under low light. Treatment with photosynthetic inhibitors indicated that PGRL1A redox state control depends on the penultimate Fd redox state. Our results showed that redox state changes in PGRL1A are crucial to optimize photosynthesis.

Crucial role of the PTOX and CET pathways in optimizing ATP synthesis in mesophyll chloroplasts of C3 and C4 plants

Besides the linear electron flow (LET), several regulatory and alternative electron transfer pathways exist in chloroplasts, among them the cyclic electron transport around photosystem I (CET) and PTOX enzyme which oxidizes the plastoquinone pool. Until now has not been investigated the influence of PTOX activity on the regulation of CET pathways and hence the regulation of ATP production. The present study reveals differential responses of CET and PTOX regulation to environmental changes and how these pathways can adjust the ATP content to C3 and C4 metabolism. Here, we conducted a comparative study in maize (C4) and pea (C3) chloroplasts by measuring the activity of PSI in the presence of the inhibitors antimycin A and propyl gallate, estimating the content of thylakoid proteins that participate in CET, and determining ATP content and ATP synthase activity, and fluorescence analysis. We found that NDH-CET pathway plays an important role in maize chloroplasts, whereas PGR5/PGRL1 proteins play a dominant role in pea chloroplasts. The obtained results indicate a substantial involvement of PTOX activity in regulating electron flow in response to ATP demand. We also found that C3 and C4 chloroplasts contained quantitatively different CET membrane components. Based on the study findings, we propose a model of regulation of the linear electron transport (LET) and CET pathways in mesophyll chloroplasts of C4 and C3 plants under low light intensity based on ATP level and PTOX activity. • The NDH complex and PGR5/PGRL1 play a main role in CET in maize and pea mesophyll chloroplasts, respectively. • High PTOX activity in maize seems to be the key for producing ATP required for C4 metabolism. • PTOX is localized mainly in grana in maize and in stroma membranes in pea chloroplasts. • Antenna aggregation in maize indicates a high reduction, which may regulate ATP/NADPH demand.

Pulse-opencircuit voltammetry: A novel method characterizes bioanode performance from microbe-electrode interfacial processes

Bioanode is a key component of bioelectrochemical systems, but the methods characterizing its resistance distribution are lacked. We propose a novel pulse-opencircuit voltammetry (POV) based on the analytical principle clarified from the electron flow pathways of microbe-electrode interfacial processes (MEIPs). A dual-cathode cell is designed to provide an experimental platform for ensuring precise data acquisition of bioanodes. This POV method enables to measure steady state polarization curves and ohmic potential loss curves by integrating potentiostatic discharge and current interruption techniques. They determines reaction resistance (RB,act) and ohmic resistance (RB,ohm) of biofilm with the assistance of impedance spectroscopy measuring material resistance. The results of various bioanodes demonstrate that RB,act is the principal limiting factor and its value relies on catabolism state. Whilst RB,ohm is relevant to extracellular electron transfer behaviors. They are two useful indicators of the dynamic evaluation of biofilm. We anticipate that this method together with the cell platform is accessible to users and has wide applications in bioanode construction and electroactive bacteria investigation.

Retraction Note: The highly efficient NDH-dependent photosystem I cyclic electron flow pathway in the marine angiosperm Zostera marina.

Retraction Note: The highly efficient NDH-dependent photosystem I cyclic electron flow pathway in the marine angiosperm Zostera marina.

Characterization of the Wave Phenomenon in Flash-Induced Fluorescence Relaxation and Its Application to Study Cyclic Electron Pathways in Microalgae.

Photosynthesis is a series of redox reactions, in which several electron transport processes operate to provide the energetic balance of light harvesting. In addition to linear electron flow, which ensures the basic functions of photosynthetic productivity and carbon fixation, alternative electron transport pathways operate, such as the cyclic electron flow (CEF), which play a role in the fine tuning of photosynthesis and balancing the ATP/NADPH ratio under stress conditions. In this work, we characterized the electron transport processes in microalgae species that have high relevance in applied research and industry (e.g., Chlorella sorokiniana, Haematococcus pluvialis, Dunaliella salina, Nannochloropsis sp.) by using flash-induced fluorescence relaxation kinetics. We found that a wave phenomenon appeared in the fluorescence relaxation profiles of microalgae to different extents; it was remarkable in the red cells of H. pluvialis, D. salina and C. sorokiniana, but it was absent in green cells of H. pluvialis and N. limnetica. Furthermore, in microalgae, unlike in cyanobacteria, the appearance of the wave required the partial decrease in the activity of Photosystem II, because the relatively high Photosystem II/Photosystem I ratio in microalgae prevented the enhanced oxidation of the plastoquinone pool. The wave phenomenon was shown to be related to the antimycin A-sensitive pathway of CEF in C. sorokiniana but not in other species. Therefore, the fluorescence wave phenomenon appears to be a species-specific indicator of the redox reactions of the plastoquinone pool and certain pathways of cyclic electron flow.

Limiting steps and the contribution of alternative electron flow pathways in the recovery of the photosynthetic functions after freezing-induced desiccation of Haberlea rhodopensis

Haberlea rhodopensis Friv. is unique with its ability to survive desiccation to an air-dry state during periods of extreme drought and freezing temperatures. To understand its survival strategies, it is important to examine the protective mechanisms not only during desiccation but also during rehydration. We investigated the involvement of alternative cyclic electron pathways during the recovery of photosynthetic functions after freezing-induced desiccation. Using electron transport inhibitors, the role of PGR5-dependent and NDH-dependent PSI-cyclic electron flows and plastid terminal oxidase were assessed during rehydration of desiccated leaves. Recovery of PSII and PSI, the capacity of PSI-driven cyclic electron flow, the redox state of plastoquinone pool, and the intersystem electron pool were analyzed. Data showed that the effect of alternative flows is more pronounced in the first hours of rehydration. In addition, the NDH-dependent cyclic pathway played a more determining role in the recovery of PSI than in the recovery of PSII.

A species by any other name would sound as sweet.

David Smith argues against the prevalent practice of naming newly discovered species after human beings.

Self-encapsulated enzyme through in-situ growth of polypyrrole for high-performance enzymatic biofuel cell

Lacking of electron flow pathway within enzymes, enzymatic biofuel cells (EBFCs) always suffer from low power and poor stability, which limits their further application. Aiming to improve enzyme’s conductivity and stability, we constructed a conductive enzyme nanocapsule n(GOx-PPy). A self-encapsulation was triggered by adding glucose to initiate an in-situ growth of PPy network within and around the glucose oxidase (GOx). Conductive pathways, as well as protective shell, were formed between single GOx active site and PPy network, allowing enhanced electron transfer and enzymatic stability. The n(GOx-PPy)-based EBFC demonstrated 245-fold increased power output, providing a promising strategy for constructing high-performance EBFCs.

Hollow InVO4 Nanocuboid Assemblies toward Promoting Photocatalytic N2 Conversion Performance.

The unique InVO4 mesocrystal superstructure, particularly with cubical skeleton and hollow interior, which consists of numerous nanocube building blocks, closely stacking by stacking, aligning by aligning, and sharing the same crystallographic orientations, is successfully fabricated. The synergy of a reaction-limited aggregation and an Ostwald ripening process is reasonably proposed for the growth of this unique superstructure. Both single-particle surface photovoltage and confocal fluorescence spectroscopy measurements demonstrate that the long-range ordered mesocrystal superstructures can significantly retard the recombination of electron-hole pairs through the creation of a new pathway for anisotropic electron flow along the inter-nanocubes. This promising charge mobility feature of the superstructure greatly contributes to the pronounced photocatalytic performance of the InVO4 mesocrystal toward fixation of N2 into NH3 with the quantum yield of 0.50% at wavelength of 385nm.

Cooperation of chloroplast ascorbate peroxidases and proton gradient regulation 5 is critical for protecting Arabidopsis plants from photo-oxidative stress.

High-light (HL) stress enhances the production of H2 O2 from the photosynthetic electron transport chain in chloroplasts, potentially causing photo-oxidative damage. Although stromal and thylakoid membrane-bound ascorbate peroxidases (sAPX and tAPX, respectively) are major H2 O2 -scavenging enzymes in chloroplasts, their knockout mutants do not exhibit a visible phenotype under HL stress. Trans-thylakoid proton gradient (∆pH)-dependent mechanisms exist for controlling H2 O2 production from photosynthesis, such as thermal dissipation of light energy and downregulation of electron transfer between photosystems II and I, and these may compensate for the lack of APXs. To test this hypothesis, we focused on a proton gradient regulation 5 (pgr5) mutant, wherein both ∆pH-dependent mechanisms are impaired, and an Arabidopsis sapx tapx double mutant was crossed with the pgr5 single mutant. The sapx tapx pgr5 triple mutant exhibited extreme sensitivity to HL compared with its parental lines. This phenotype was consistent with cellular redox perturbations and enhanced expression of many oxidative stress-responsive genes. These findings demonstrate that the PGR5-dependent mechanisms compensate for chloroplast APXs, and vice versa. An intriguing finding was that the failure of induction of non-photochemical quenching in pgr5 (because of the limitation in ∆pH formation) was partially recovered in sapx tapx pgr5. Further genetic studies suggested that this recovery was dependent on the NADH dehydrogenase-like complex-dependent pathway for cyclic electron flow around photosystem I. Together with data from the sapx tapx npq4 mutant, we discuss the interrelationship between APXs and ∆pH-dependent mechanisms under HL stress.

The highly efficient NDH-dependent photosystem I cyclic electron flow pathway in the marine angiosperm Zostera marina.

Zostera marina, a fully submerged marine angiosperm with a unique evolutionary history associated with its terrestrial origin, has distinct photochemical characteristics caused by its oxygen-evolving complex (OEC) being prone to deactivation in visible light. Based on the present phylogenetic analysis, the chloroplast NADPH dehydrogenase-like (NDH) complex was found to be completed in of Z. marina, unlike other marine plants, suggesting its crucial role. Thus, the responses of electron transport to irradiation were investigated through multiple chlorophyll fluorescence techniques and Western blot analysis. Moreover, the respective contribution of the two photosystem I cyclic electron flow (PSI-CEF) pathways to the generation of trans-thylakoid proton gradient (∆pH) was also examined using inhibitors. The contributions of the two PSI-CEF pathways to ∆pH were similar; furthermore, there was a trade-off between the two pathways under excess irradiation: the PGR5/L1-dependent PSI-CEF decreased gradually following its activation during the initial illumination, while NDH-dependent PSI-CEF was activated gradually with exposure duration. OEC inactivation was continuously under excess irradiation, which exhibits a positive linear correlation with the activation of NDH-dependent PSI-CEF. We suggest that PGR5/L1-dependent PSI-CEF was preferentially activated to handle the excess electron caused by the operation of OEC during the initial illumination. Subsequently, the increasing OEC inactivation with exposure duration resulted in a deficit of electrons. Limited electrons from PSI might preferentially synthesize NADPH, which could support the function of NDH-dependent PSI-CEF to generate ∆pH and ATP via reducing ferredoxin, thereby maintaining OEC stability.

Regulation of photosynthetic cyclic electron flow pathways by adenylate status in higher plant chloroplasts

Cylic electron flow (CEF) around Photosystem I in photosynthetic eukaryotes is likely to be necessary to augment ATP production, rapidly- and precisely balancing the plastid ATP/NADPH energy budget to meet the demands of downstream metabolism. Many regulatory aspects of this process are unclear. Here we demonstrate that the higher plant plastid NADH/Fd:plastoquinone reductase (NDH) and proposed PGR5/PGRL1 ferredoxin:plastoquinone reductase (FQR) pathways of CEF are strongly, rapidly and reversibly inhibited in vitro by ATP with Ki values of 670 μM and 240 μM respectively, within the range of physiological changes in ATP concentrations. Control experiments ruled out effects on secondary reactions, e.g. FNR- and cytochrome b6f activity, nonphotochemical quenching of chlorophyll fluorescence etc., supporting the view that ATP is an inhibitor of CEF and its associated pmf generation and subsequent ATP production. The effects are specific to ATP, with the ATP analog AMP-PNP showing little inhibitory effect, and ADP inhibiting only at higher concentrations. For the FQR pathway, inhibition was found to be classically competitive with Fd, and the NDH pathway showing partial competition with Fd. We propose a straightforward model for regulation of CEF in plants in which CEF is activated under conditions when stromal ATP low, but is downregulated as ATP levels build up, allowing for effective ATP homeostasis. The differences in Ki values suggest a two-tiered regulatory system, where the highly efficient proton pumping NDH is activated with moderate decreases in ATP, with the less energetically-efficient FQR pathway being activated under more severe ATP depletion.

Deconvolution of reduction potentials of formate dehydrogenase from Cupriavidus necator.

The formate dehydrogenase enzyme from Cupriavidus necator (FdsABG) carries out the two-electron oxidation of formate to CO2, but is also capable of reducing CO2 back to formate, a potential biofuel. FdsABG is a heterotrimeric enzyme that performs this transformation using nine redox-active cofactors: a bis(molybdopterin guanine dinucleotide) (bis-MGD) at the active site coupled to seven iron-sulfur clusters, and one equivalent of flavin mononucleotide (FMN). To better understand the pathway of electron flow in FdsABG, the reduction potentials of the various cofactors were examined through direct electrochemistry. Given the redundancy of cofactors, a truncated form of the FdsA subunit was developed that possesses only the bis-MGD active site and a singular [4Fe-4S] cluster. Electrochemical characterization of FdsABG compared to truncated FdsA shows that the measured reduction potentials are remarkably similar despite the truncation with two observable features at - 265mV and - 455mV vs SHE, indicating that the voltammetry of the truncated enzyme is representative of the reduction potentials of the intact heterotrimer. By producing truncated FdsA without the necessary maturation factors required for bis-MGD insertion, a form of the truncated FdsA that possesses only the [4Fe-4S] was produced, which gives a single voltammetric feature at -525mV, allowing the contributions of the molybdenum cofactor to be associated with the observed feature at - 265mV. This method allowed for the deconvolution of reduction potentials for an enzyme with highly complex cofactor content to know more about the thermodynamic landscape of catalysis.

Proteomic responses to ocean acidification of the marine diazotroph Trichodesmium under iron-replete and iron-limited conditions.

Growth and dinitrogen (N2) fixation of the globally important diazotrophic cyanobacteria Trichodesmium are often limited by iron (Fe) availability in surface seawaters. To systematically examine the combined effects of Fe limitation and ocean acidification (OA), T. erythraeum strain IMS101 was acclimated to both Fe-replete and Fe-limited concentrations under ambient and acidified conditions. Proteomic analysis showed that OA affected a wider range of proteins under Fe-limited conditions compared to Fe-replete conditions. OA also led to an intensification of Fe deficiency in key cellular processes (e.g., photosystem I and chlorophyll a synthesis) in already Fe-limited T. erythraeum. This is a result of reallocating Fe from these processes to Fe-rich nitrogenase to compensate for the suppressed N2 fixation. To alleviate the Fe shortage, the diazotroph adopts a series of Fe-based economic strategies (e.g., upregulating Fe acquisition systems for organically complexed Fe and particulate Fe, replacing ferredoxin by flavodoxin, and using alternative electron flow pathways to produce ATP). This was more pronounced under Fe-limited-OA conditions than under Fe limitation only. Consequently, OA resulted in a further decrease of N2- and carbon-fixation rates in Fe-limited T. erythraeum. In contrast, Fe-replete T. erythraeum induced photosystem I (PSI) expression to potentially enhance the PSI cyclic flow for ATP production to meet the higher demand for energy to cope with the stress caused by OA. Our study provides mechanistic insight into the holistic response of the globally important N2-fixing marine cyanobacteria Trichodesmium to acidified and Fe-limited conditions of future oceans.

Role of cyclic and pseudo-cyclic electron transport in response to dynamic light changes in Physcomitrella patens.

Photosynthetic organisms support cell metabolism by harvesting sunlight and driving the electron transport chain at the level of thylakoid membranes. Excitation energy and electron flow in the photosynthetic apparatus is continuously modulated in response to dynamic environmental conditions. Alternative electron flow around photosystem I plays a seminal role in this regulation contributing to photoprotection by mitigating overreduction of the electron carriers. Different pathways of alternative electron flow coexist in the moss Physcomitrella patens, including cyclic electron flow mediated by the PGRL1/PGR5 complex and pseudo-cyclic electron flow mediated by the flavodiiron proteins FLV. In this work, we generated P.patens plants carrying both pgrl1 and flva knock-out mutations. A comparative analysis of the WT, pgrl1, flva, and pgrl1 flva lines suggests that cyclic and pseudo-cyclic processes have a synergic role in the regulation of photosynthetic electron transport. However, although both contribute to photosystem I protection from overreduction by modulating electron flow following changes in environmental conditions, FLV activity is particularly relevant in the first seconds after a light change whereas PGRL1 has a major role upon sustained strong illumination.

Electrical Conductivity Modulation of Crosslinked Composite Nanofibers Based on PEO and PEDOT:PSS

The aim of this work is to investigate the development of nanofiber mats, based on intrinsically conductive polymers (ICPs), which show simultaneously a high electrical conductivity and mandatory insoluble water properties. In particular, the nanofibers, thanks to their properties such as high surface area, porosity, and their ability to offer a preferential pathway for electron flow, play a crucial role to improve the essential characteristics ensured by ICPs. The nanofiber mats are obtained by electrospinning process, starting from a polymeric solution made of polyethylene oxide (PEO) and poly(styrene sulfonate) (PEDOT:PSS). PEO is selected not only as a dopant to increase the electrical/ionic conductivity, as deeply reported in the literature, but also to ensure the proper stability of the polymeric jet, to collect a dried nanofiber mat. Moreover, in the present work, two different treatments are proposed in order to induce crosslinking between PEO chains and PEDOT:PSS, made insoluble into water which is the final sample. The first process is based on a heating treatment, conducted at 130°C under nitrogen atmosphere for 6 h, named the annealing treatment. The second treatment is provided by UV irradiation that is effective to induce a final crosslinking, when a photoinitiator, such as benzophenone, is added. Furthermore, we demonstrate that both crosslinking treatments can be used to verify the preservation of nanostructures and their good electrical conductivity after water treatment (i.e., water resistance). In particular, we confirm that the crosslinking method with UV irradiation results to being more effective than the standard annealing treatment. Indeed, we demonstrate that the processing time, required to obtain the final crosslinked nanofiber mats with a high electrical conductance, results to being smaller than the one needed during the heating treatment.