Electron Donor Substrates Research Articles

Fast Reaction of the Prussian Blue Based Nanozyme "Artificial Peroxidase" with the Substrates: Pre-Steady-State Kinetic Approach.

This letter introduces the pre-steady-state kinetic approach, which is traditional for evaluation of elementary constants in molecular (enzyme) catalysis, for nanozymes. Apparently, the most active peroxidase-mimicking nanozyme based on catalytically synthesized Prussian Blue nanoparticles has been chosen. The elementary constants (k1) for the nanozymes' reduction by an electron-donor substrate (being the fastest stage according to steady-state kinetic data) have been determined by means of stopped-flow spectroscopy. These constants have been found to be dependent on both the size of the nanozyme and the reducing substrate redox potential. For the smallest nanozymes (32 nm in diameter), log(k1) linearly decays with an increase of the substrate redox potential (cotangent value ≈125 mV). On the contrary, for the largest nanozymes with a diameter above 150 nm, k1 is almost independent of it. Moreover, for the substrate with the lowest redox potential (K4[Fe(CN)6]), the rate constant under discussion (k1) is almost independent of the nanozymes' size. Perhaps, the rate of the intrananozyme electron transfer causing bleaching becomes comparative or even lower than that of the nanoparticle interaction with the fastest substrate. Anyway, the elementary constant of nanozyme reduction with potassium ferrocyanide (k1) reaches the value of 1 × 1010 M-1 s-1, which is 3-4 orders of magnitude faster than for enzymes peroxidases. The obtained results obviously demonstrate that the pre-steady-state kinetic approach is able to discover novel advantages of nanozymes from both fundamental and practical points of view.

Distinct roles of biochar and pyrite substrates in enhancing nutrient and heavy metals removal in intermittent-aerated constructed wetlands: Performances and mechanism

Constructed wetlands have been widely employed as a cost-effective and environmentally friendly alternative for treating primary and secondary sewage effluents. In this study, biochar and pyrite were utilized as electron donor substrates in intermittent-aerated vertical flow constructed wetlands to strengthen the nutrient and heavy metals removal simultaneously, and the response of nutrient reduction and microbial community to heavy metals stress was also explored. The results indicated that biochar addition exhibited a better nitrogen removal, while pyrite addition greatly promoted the phosphorus removal. Moreover, the high removal efficiencies of Cu2+, Pb2+ and Cd2+ (above 90%) except for Zn2+ were obtained in each system. However, the exposure of heavy metals decreased phosphorus removal while had little effect on nitrogen removal. The influent load and intermittent aeration implementation led to a significant shift in microbial community structures, but microbial biodiversity and abundance decreased under the exposure of heavy metals. Particularly, Thiobacillus and Ferritrophicum, associated with sulfur autotrophic denitrification and iron autotrophic denitrification, were more abundant in pyrite-based wetland systems.

The role of graphene in SnS2@Graphene for rechargeable lithium batteries: A view from the electronic structure

Based on the first-principles study, the adsorption and electron transfer properties of Li atom at different sites of SnS2 monolayer, SnS2@Graphene 2D-nanocomposite are analyzed. The differential charge density and density of states (DOS) analysis show that the graphene substrate as an electron donor can change the 2D-nanocomposite from a semiconductor to a metal, and reduce the adsorption energy of Li atom by decreasing the charge transferring from Li atom to SnS2. This indicates that graphene substrate is beneficial for improving the performance of SnS2@Graphene. Meanwhile, the Li atoms tend not to cluster on the SnS2@Graphene 2D-nanocomposite, which is useful to prolong the lifespan of the SnS2@Graphene. The functionality of graphene in SnS2@Graphene 2D-nanocomposite is proved by other electron donor substrates, such as a two-H-atom model and a Sn (111) substrate model. All the results indicate that the graphene, as an electron donor in SnS2@Graphene 2D-nanocomposite, plays a key role in improving the performance of SnS2 in rechargeable lithium batteries.

Enhancement of immobilized biochar/FeS on nitrogen removal in constructed wetland at low temperature

Constructed wetland (CW) is an eco-friendly and economically treatment for the deep nitrogen removal. Microbial denitrification, a key process in CW, is temperature-sensitive, with decreased nitrogen removal efficiency at low temperature. To enhance the nitrogen removal efficiency at low temperature, this study explored the use of functional substrates, focusing on iron sulfide (FeS) nanoparticles and biochar (BC). The results showed that immobilized BC/FeS significantly improved nitrate (NO3-N) and total nitrogen (TN) removal in CW at low temperature (8–13 °C), which reached 93.59 ± 2.07 and 80.06 ± 3.00 %, respectively, with 30 % immobilized BC/FeS. Increasing the dosage of immobilized BC/FeS accelerated denitrification rate and reduced nitrite (NO2-N) accumulation. Autotrophic denitrification was the primary pathway, with the contribution of autotrophic denitrification increasing from 64.06 % (10 % immobilized BC/FeS) to 83.25 % (30 % immobilized BC/FeS). Sulfurimonas was the dominant denitrifying bacteria identified as a cold-tolerant denitrifying bacteria. The study underlines the potential of composite electron donor substrates to enhance nitrogen removal in CW, providing a sustainable solution to nitrogen pollution in cold climates.

A new versatile peroxidase with extremophilic traits over-produced in MicroTom cell cultures

Peroxidases are widespread key antioxidant enzymes that catalyse the oxidation of electron donor substrates in parallel with the decomposition of H2O2. In this work, a novel tomato peroxidase, named SAAP2, was isolated from MicroTom cell cultures, purified, and characterised. The enzyme was identified with 64% sequence coverage as the leprx21 gene product (suberization-associated anionic peroxidase 2-like) from Solanum lycopersicum, 334 amino acids long. Compared to other plant peroxidases, SAAP2 was more active at elevated temperatures, with the optimal temperature and pH at 90 °C and 5.0, respectively. Furthermore, the enzyme retained more than 80% of its maximal activity over the range of 70–80 °C and the presence of NaCl (1.0–4.5 M). It also exhibited broad pH versatility (65% relative activity over the pH range 2.0–7.0), acid-tolerance (80% residual activity after 22 h at pH 2.0–7.0), high thermostability (50% residual activity after 2 h at 80 °C) and proteolytic resistance. SAAP2 exhibited exceptional resistance under thermo-acidic conditions compared to the horseradish peroxidase benchmark, suggesting that it may find potential applications as a supplement or anti-pollution agent in the food industry.

Charge-Transfer-Driven Phase Transition of Two-Dimensional MoTe2 in Donor-Acceptor Heterostructures.

In this work, based on first-principles calculations, we propose that electrene can be considered as an electron-donating substrate to drive the phase transition of MoTe2 from the H to T' phase, which is a topic of long-standing interest and importance. In particular, new electrenes Ca2XN2 (X = Zr, Hf) are predicted with the existence of a nearly free two-dimensional (2D) electron gas and ultralow work functions. In MoTe2/Ca2XN2 donor-acceptor heterostructures, we find significantly large charge transfer (∼0.4e per MoTe2 unit cell) from Ca2XN2 to MoTe2, which stabilizes the T' phase and decreases the phase transition barrier (from ∼0.9 to ∼0.5 eV per unit cell). In addition, the phase transition of MoTe2 on Ca2XN2 remains effective as the interlayer distance varies. It therefore can be confirmed conclusively that our results open a new avenue for phase transition study and provide new insights for the large-scale synthesis of metastable high-quality T'-phase MoTe2.

Evaluation of the decolorization potential of azo dyes by aerobic granular sludge

This study assessed the azo dye decolorization potential by aerobic granular sludge (AGS), particularly investigating the effects of different electron donor substrates, redox mediators, dye structures, and alternative electron acceptors. Additionally, the stability and performance of an AGS reactor were evaluated during the treatment of a synthetic azo dye-containing effluent in the absence and presence of a redox mediator. Propionate and anthraquinone-2-sulfonate (AQS) were the best substrate (k1 1.1–1.9-fold higher) and redox mediator (k1 1.4-fold higher), respectively, for Reactive Black 5 (RB5) decolorization. AQS (50 μM) had a greater impact on Reactive Orange 16 (k1 = 15.60 h−1) than on Reactive Red 120 (k1 = 0.48 h−1) and RB5 (k1 = 7.92 h−1). Nitrate and sulfate (100 and 200 mg·L−1) inhibited dye reduction (decreasing k1 from 5.52 to 0.72 and 2.40 h−1, respectively), possibly because they compete with the dye for the electrons in the environment. However, AQS minimized their impact on this process (except for nitrate at 200 mg·L−1). In the reactor, RB5 and AQS did not alter the physical characteristics of AGS and its ability to remove organic matter and nutrients. Finally, the effect of AQS on decolorization was not so intense, increasing only ∼15%.

ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates.

In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self-sustaining cascade reached a titer of (S)-3-hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi-functional ones.

ATP‐Independent and Cell‐Free Biosynthesis of β‐Hydroxy Acids Using Vinyl Esters as Smart Substrates

AbstractIn vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β‐hydroxy acids through a non‐decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self‐sustaining cascade reached a titer of (S)‐3‐hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi‐functional ones.

Trialkylphosphonium oxoborates as C(sp3)-H oxyanion holes and their application in catalytic chemoselective acetalization.

The use of trialkylphosphonium oxoborates (TOB) as catalysts is reported. The site-isolated borate counter anion in a TOB catalyst increases the availability of C(sp3)-H to interact with electron donor substrates. The catalytic protocol is applicable to a wide range of substrates in the acetalization reaction and provides excellent chemoselectivity in the acetalization over thioacetalization in the presence of alcohols and thiols, which is otherwise hard to achieve using typical acid catalysts. Experimental and computational studies revealed that the TOB catalysts have multiple preorganized C(sp3)-Hs that serve as a mimic of oxyanion holes, which can stabilize the oxyanion intermediates via multiple C(sp3)-H non-classical hydrogen bond interactions.

Biochemical Characterization of Pyranose Oxidase from Streptomyces canus-Towards a Better Understanding of Pyranose Oxidase Homologues in Bacteria.

Pyranose oxidase (POx, glucose 2-oxidase; EC 1.1.3.10, pyranose:oxygen 2-oxidoreductase) is an FAD-dependent oxidoreductase and a member of the auxiliary activity (AA) enzymes (subfamily AA3_4) in the CAZy database. Despite the general interest in fungal POxs, only a few bacterial POxs have been studied so far. Here, we report the biochemical characterization of a POx from Streptomyces canus (ScPOx), the sequence of which is positioned in a separate, hitherto unexplored clade of the POx phylogenetic tree. Kinetic analyses revealed that ScPOx uses monosaccharide sugars (such as d-glucose, d-xylose, d-galactose) as its electron-donor substrates, albeit with low catalytic efficiencies. Interestingly, various C- and O-glycosides (such as puerarin) were oxidized by ScPOx as well. Some of these glycosides are characteristic substrates for the recently described FAD-dependent C-glycoside 3-oxidase from Microbacterium trichothecenolyticum. Here, we show that FAD-dependent C-glycoside 3-oxidases and pyranose oxidases are enzymes belonging to the same sequence space.

Bilirubin oxidase as a single enzymatic oxygen scavenger for the development of reductase-based biosensors in the open air and its application on a nitrite biosensor

The commercialization of amperometric or voltammetric biosensors that operate at potentials lower than −0.2 V vs SHE has been hindered by the need for anoxic working conditions due to the interference of molecular oxygen, whose electrochemical reduction can potentially mask other redox processes and generate reactive oxygen species (ROS). A deoxygenation step must be thus integrated into the analytical process. To this end, several (bio)chemical oxygen scavenging systems have been proposed, such as the bi-enzyme system, glucose oxidase/catalase. Still, a few issues persist owing to enzyme impurities and the formation of oxygen reactive species.Here in, we propose a new mono-enzymatic oxygen scavenging system composed of a multicopper oxidase as a single biocatalytic oxygen reducer. As a model, we used bilirubin oxidase (BOD), which catalyzes the direct reduction of oxygen to water in the presence of an electron donor substrate, without releasing hydrogen peroxide. Both the direct electron transfer and mediated electrochemical approach using different co-substrates were screened for the ability to promote the enzymatic reduction of oxygen. An optimal combination of BOD with sodium ascorbate proved to be quick (5 min) and effective. It was subsequently employed, as a proof-of-concept, in a voltammetric biosensor based on a multiheme cytochrome c nitrite reductase, which performs the reduction of nitrite to ammonia at potentials below −0.3 V vs SHE. The nitrite biosensor performed well under ambient air, with no need for a second enzyme to account for the build-up of oxygen reactive intermediaries.

Nanostructured Surfaces with Multimodal Antimicrobial Action.

Self-disinfecting surfaces are a current pressing need for public health and safety in view of the current COVID-19 pandemic, where the keenly felt worldwide repercussions have highlighted the importance of infection control, frequent disinfection, and proper hygiene. Because of its potential impact upon real-world translation into downstream applications, there has been much research interest in multiple disciplines such as materials science, chemistry, biology, and engineering. Various antimicrobial technologies have been developed and currently applied on surfaces in public spaces, such as elevator buttons and escalator handrails. These technologies are mainly based on conventional methods of grafting quaternary ammonium salts (QACs) such as benzalkonium chloride or the immobilization of metal species of silver or copper. However, neither the long-term efficacy nor the fast-killing properties have been proven, and the future repercussions from extended use, such as environmental hazards and the induction of MDR development, is unknown. Nanostructured surfaces with excellent antimicrobial activities have been claimed to be the next generation of self-disinfecting surfaces with various promising applications and passive antimicrobial mechanisms, without the potential repercussions of active ingredient overuse. In this Account, we briefly introduce the concept of mechanobactericidal action realized by these nanostructured surfaces first discovered on cicada wings. The elimination of microbes on the surface was actualized by the physical rupture of the microbe cell wall by nanoprotusions, without any involvement of chemical species. By mimicking the physical features of naturally occurring biocidal surfaces, the fabrication of nanostructures on various substrates such as titania, silicon, and polymers has been well described. Observations of the dependence of their antimicrobial efficacy on physical characteristics such as height, density, and rigidity have also been documented. However, the complex fabrication of such nanostructures remains the main drawback preventing its widespread application. We outline our efforts in fabricating a series of zinc-based nanostructured materials with facile and generally applicable wet chemistry methods, including nanodaggered zeolitic imidazolate frameworks (ZIF-L) and ZnO nanoneedles. In our investigations, we discovered that there were additional modes of action that contributed to the excellent biocidal activities of our materials. The impact of surface chemistry and charge was partially responsible for the selectivity and efficacy of ZIF-L-coated surfaces, where the positively charged surfaces were able to attract and adhere negatively charged bacteria to the surface. The combination of semiconductor ZnO nanoneedles on electron-donating substrates allowed for the generation of reactive oxygen species (ROS), realizing the remote killing of bacteria unadhered to the nanostructured surface. Additionally, we demonstrate several real-life applications of the synthesized materials, underscoring the importance of materials development suited for scale-up and eventual translation to potential applications and commercial end products.

A TiO2 -Co(terpyridine)2 Photocatalyst for the Selective Oxidation of Cellulose to Formate Coupled to the Reduction of CO2 to Syngas.

Immobilization of a phosphonated cobalt bis(terpyridine) catalyst on TiO2 nanoparticles generates a photocatalyst that allows coupling aqueous CO2 -to-syngas (CO and H2 ) reduction to selective oxidation of biomass-derived oxygenates or cellulose to formate. An enzymatic saccharification pre-treatment process is employed that enables the use of insoluble cellulose as an electron-donating substrate under benign aqueous conditions suitable for photocatalytic CO2 conversion. The hybrid photocatalyst consists of solely earth-abundant components, and its heterogeneous nature allows for reuse and operation in aqueous solution for several days at 25 °C, reaching a cellulose-to-formate conversion yield of 17 %. Thus, the proof-of-concept for valorizing two waste streams (CO2 and biomass) simultaneously into value-added chemicals through solar-driven catalysis is demonstrated.

Bacteria Make a Living Breathing the Nitroheterocyclic Insensitive Munitions Compound 3-Nitro-1,2,4-triazol-5-one (NTO).

The nitroheterocyclic 3-nitro-1,2,4-triazol-5-one (NTO) is an ingredient of insensitive explosives increasingly used by the military, becoming an emergent environmental pollutant. Cometabolic biotransformation of NTO occurs in mixed microbial cultures in soils and sludges with excess electron-donating substrates. Herein, we present the unusual energy-yielding metabolic process of NTO respiration, in which the NTO reduction to 3-amino-1,2,4-triazol-5-one (ATO) is linked to the anoxic acetate oxidation to CO2 by a culture enriched from municipal anaerobic digester sludge. Cell growth was observed simultaneously with NTO reduction, whereas the culture was unable to grow in the presence of acetate only. Extremely low concentrations (0.06 mg L-1) of the uncoupler carbonyl cyanide m-chlorophenyl hydrazone inhibited NTO reduction, indicating that the process was linked to respiration. The ultimate evidence of NTO respiration was adenosine triphosphate production due to simultaneous exposure to NTO and acetate. Metagenome sequencing revealed that the main microorganisms (and relative abundances) were Geobacter anodireducens (89.3%) and Thauera sp. (5.5%). This study is the first description of a nitroheterocyclic compound being reduced by anaerobic respiration, shedding light on creative microbial processes that enable bacteria to make a living reducing NTO.

Exploration of processes governing microbial reductive dechlorination in a heterogeneous aquifer flow cell

Although microbial reductive dechlorination (MRD) has proven to be an effective approach for in situ treatment of chlorinated ethenes, field implementation of this technology is complicated by many factors, including subsurface heterogeneity, electron donor availability, and distribution of microbial populations. This work presents a coupled experimental and mathematical modeling study designed to explore the influence of heterogeneity on MRD and to assess the suitability of microcosm-derived rate parameters for modeling complex heterogeneous systems. A Monod-based model is applied to simulate a bioremediation experiment conducted in a laboratory-scale aquifer cell packed with aquifer material from the Commerce Street Superfund site in Williston, VT. Results reveal that (uncalibrated) model application of microcosm-derived dechlorination and microbial growth rates for transformation of trichloroethene (TCE), cis-1,2-dichloroethene (cis-DCE), and vinyl chloride (VC) reproduced observed aquifer cell concentration levels and trends. Mean relative errors between predicted and measured effluent concentrations were quantified as 6.7%, 27.0%, 41.5%, 32.0% and 21.6% over time for TCE, cis-DCE, VC, ethene and total volatile fatty acids (fermentable electron donor substrate and carbon source), respectively. The time-averaged extent of MRD (i.e., ethene formation) was well-predicted (4% underprediction), with modeled MRD exhibiting increased deviation from measured values under electron donor limiting conditions (maximum discrepancy of 14%). In contrast, simulations employing a homogeneous (uniform flow) domain resulted in underprediction of MRD extent by an average of 13%, with a maximum discrepancy of 45%. Model sensitivity analysis suggested that trace amounts of natural dissolved organic carbon served as an important fermentable substrate, providing up to 69% of the reducing equivalents consumed for MRD under donor-limiting conditions. Aquifer cell port concentration data and model simulations revealed that ethene formation varied spatially within the domain and was associated with regions of longer residence times. These results demonstrate the strong influence of subsurface heterogeneity on the accuracy of MRD predictions, and highlight the importance of subsurface characterization and the incorporation of flow field uncertainty in model applications for successful design and assessment of in situ bioremediation.

Electricity generation using membrane‐less microbial fuel cell powered by sludge supplemented with lignocellulosic waste

A membrane-less microbial fuel cell (ML-MFC) is an electrochemical device that incorporates microorganisms into the design in order to produce electricity through biologically catalyzed oxidation of soluble, electron-donating substrates. In this study, three lignocellulosic raw materials were added into the ML-MFC whereby the sludge acted as the pseudomembrane. All three materials were used as the substrates in ML-MFC for the production of electricity that was measured using a digital multimeter. Results showed that the ML-MFC that contained sludge supplemented with banana peel produced the highest electricity, followed by corn bran and palm oil mill effluent (POME) at 237.1 mV (23.75 mW/m2), 176.8 mV (12.65 mW/m2), and 138 mV (22.03 mW/m2) after 138 h, 192 h, and 108 h of incubation period, respectively. For the control test (sludge only), about 162.7 mV was recorded at shorter incubation period (84 h). This showed that long-term operation of the ML-MFC using these complex lignocellulosic compounds as a direct substrate for electricity generation is feasible, though their degradation is slow.

Targeting the menaquinol binding loop of mycobacterial cytochrome bd oxidase.

Mycobacteria have shown enormous resilience to survive and persist by remodeling and altering metabolic requirements. Under stringent conditions or exposure to drugs, mycobacteria have adapted to rescue themselves by shutting down their major metabolic activity and elevate certain survival factor levels and efflux pathways to survive and evade the effects of drug treatments. A fundamental feature in this adaptation is the ability of mycobacteria to vary the enzyme composition of the electron transport chain (ETC), which generates the proton motive force for the synthesis of adenosine triphosphate via oxidative phosphorylation. Mycobacteria harbor dehydrogenases to fuel the ETC, and two terminal respiratory oxidases, an aa3-type cytochrome c oxidase (cyt-bcc-aa3) and a bacterial specific cytochrome bd-type menaquinol oxidase (cyt-bd). In this study, we employed homology modeling and structure-based virtual screening studies to target mycobacteria-specific residues anchoring the b558 menaquinol binding region of Mycobacterium tuberculosis cyt-bd oxidase to obtain a focused library. Furthermore, ATP synthesis inhibition assays were carried out. One of the ligands MQL-H2 inhibited both NADH2- and succinate-driven ATP synthesis inhibition of Mycobacterium smegmatis inside-out vesicles in micromolar potency. Similarly, MQL-H2 also inhibited NADH2-driven ATP synthesis in inside-out vesicles of the cytochrome-bcc oxidase deficient M. smegmatis strain. Since neither varying the electron donor substrates nor deletion of the cyt-bcc oxidase, a major source of protons, hindered the inhibitory effects of the MQL-H2, reflecting that MQL-H2 targets the terminal oxidase cytochrome bd oxidase, which was consistent with molecular docking studies. Characterization of novel cytochrome bd oxidase Menaquinol binding domain inhibitor (MQL-H2) using virtual screening and ATP synthesis inhibition assays.

Comparison of Direct and Mediated Electron Transfer for Bilirubin Oxidase from Myrothecium Verrucaria. Effects of Inhibitors and Temperature on the Oxygen Reduction Reaction

One of the processes most studied in bioenergetic systems in recent years is the oxygen reduction reaction (ORR). An important challenge in bioelectrochemistry is to achieve this reaction under physiological conditions. In this study, we used bilirubin oxidase (BOD) from Myrothecium verrucaria, a subclass of multicopper oxidases (MCOs), to catalyse the ORR to water via four electrons in physiological conditions. The active site of BOD, the T2/T3 cluster, contains three Cu atoms classified as T2, T3α, and T3β depending on their spectroscopic characteristics. A fourth Cu atom; the T1 cluster acts as a relay of electrons to the T2/T3 cluster. Graphite electrodes were modified with BOD and the direct electron transfer (DET) to the enzyme, and the mediated electron transfer (MET) using an osmium polymer (OsP) as a redox mediator, were compared. As a result, an alternative resting (AR) form was observed in the catalytic cycle of BOD. In the absence and presence of the redox mediator, the AR direct reduction occurs through the trinuclear site (TNC) via T1, specifically activated at low potentials in which T2 and T3α of the TNC are reduced and T3β is oxidized. A comparative study between the DET and MET was conducted at various pH and temperatures, considering the influence of inhibitors like H2O2, F−, and Cl−. In the presence of H2O2 and F−, these bind to the TNC in a non-competitive reversible inhibition of O2. Instead; Cl− acts as a competitive inhibitor for the electron donor substrate and binds to the T1 site.

Small Reorganization Energy for Ligand-Centered Electron-Transfer Reduction of Compound I to Compound II in a Heme Model Study

The electron-transfer (ET) processes from electron-donor substrates to oxoiron(IV) porphyrin π-cation-radical species (Cpd I) are key steps in their oxygenation reactions. Here, we have evaluated the rate constants of the outer-sphere ET reduction of Cpd I model complexes of meso-tetramesitylporphyrin (1) and 2,7,12,17-tetramesityl-3,8,13,18-tetramethylporphyrin (2) in light of the Marcus theory of ET to determine the ET reorganization energies (λ). The λ values of the ligand-centered ET reduction of Cpd I model complexes are much smaller than those of the metal-centered ET reduction of various oxoiron(IV) complexes. Moreover, the λ value of 1 is larger than that of 2, resulting from the difference in the nature of the a1u/a2u porphyrin π-cation-radical orbitals.