Active Site Research Articles

Cladophorol-A is an inhibitor of cyclic GMP-AMP synthase

Cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase (cGAS) is an enzyme sensor of double-stranded DNA (dsDNA) that serves to trigger activation of the cGAS-stimulator of interferon genes (STING) pathway. Excessive activation of this pathway has been demonstrated to contribute to various forms of inflammatory disease. As such, cGAS has arisen as a potential therapeutic target with broad potential applications. Using a pathway-targeted cell-based screening approach, we identified the natural product Cladophorol A as a new class of non-cytotoxic cGAS inhibitor (cell-based IC50 = 370 nM). An x-ray co-crystal structure at 2.75 Å resolution revealed that Cladophorol A inhibits cGAS by binding to its active site within the conserved adenosine nucleobase binding site.

Discovery of a new lead molecule to develop a novel class of human factor XIIa inhibitors.

Factor XIIa (FXIIa) is a plasma serine protease within the contact activation pathway. Inhibiting FXIIa could offer a viable therapeutic approach for achieving effective and safer anticoagulation without the bleeding risks that accompany the use of existing anticoagulants. Therefore, we investigated the anticoagulant properties of an amidine-containing molecule (inhibitor 1) to identify a potential lead molecule for subsequent development of FXIIa inhibitors. Results indicated that inhibitor 1 primarily inhibits human FXIIa with an IC50 value of ~30 µM. The inhibitor demonstrated variable selectivity against thrombin, factor IXa, factor Xa, factor XIa, and activated protein C. Michaelis-Menten kinetics indicated that the molecule is an active site inhibitor of FXIIa. Molecular modeling studies revealed that the molecule recognizes residues His57, Asp189, and Ala190 in FXIIa's active site. The inhibitor selectively and concentration-dependently prolonged the clotting time of human plasma under activated partial thromboplastin time assay conditions. The inhibitor did not exhibit significant cytotoxicity in human HEK293 cells and the in silico pharmacokinetics and toxicology data were comparable to known anticoagulants. This study introduces inhibitor 1 as a lead platform for further development as an anticoagulant to provide a more effective and safer approach to preventing and treating thromboembolic diseases.

Design, Synthesis, Biological and Computational Analysis of Isatin-based bis-Thiourea Analogues as Anti-diabetic and Anti-nematode Agents

A new series of isatin derivatives were synthesized, characterized by 1HNMR, 13CNMR and HREI-MS, and screened for α-glucosidase and alpha amylase inhibition. All the analogues were found to be dual inhibitors and showed good inhibitory potentials with IC50 values ranging from 5.28 ± 0.10 to 38.66 ± 0.30 µM (against alpha-amylase), and 5.45 ± 0.10 to 39.25 ± 0.50 µM (against alpha-glucosidase), as compared to the standard drug acarbose (IC50 = 11.12 ± 0.15 and 11.29 ± 0.07 µM, respectively). The most potent inhibitor among the series was analogue 24 (IC50 = 5.28 ± 0.10 for alpha-amylase and IC50 = 5.46 ± 0.10 µM for alpha-glucosidase), which has a nitro group attached at the meta-position of the phenyl ring A and the para position of phenyl ring B. Structure-activity relationship has been established mainly based on the position, nature and number of the substituent(s) attached to the phenyl ring. To investigate the binding interaction of the potent analog with the active site of an enzyme, molecular docking studies were carried out. To study the drug-likeness properties, the ADME study was also carried out. The most active compounds engage most of the amino acids composing the active site and display maximum interactions. These interactions majorly include formation of strong hydrogen bonds, which might be due to the presence of highly electronegative heteroatoms on aromatic rings. All the analogues were also tested for in vivo anti-nematodal activity against C. elegans to assess their cytotoxicity in comparison to the reference Levamisole. The cytotoxicity profile demonstrated that analogues 2, 7, 20 and 22 displayed minimum cytotoxicity at every concentration.

Sonochemical Preparation of Fe-Y Zeolite Catalyst for the Conversion of Benzene to Phenol

Enzyme-mimetic materials such as Fe-containing zeolites are of research interest due to the high selectivity for the one-step selective oxidation of benzene to phenol over the Fe-catalytic active center at room temperature and ambient pressure. This study investigates a Fe-modified Y zeolite catalysts prepared via a sonochemical method for the oxidation of benzene-to-phenol using water as solvent. These catalysts were subjected to a screening and optimization experiment by a response surface methodology (RSM), evaluating their potential for the synthesis of phenol. Achieving the preparation of a catalyst with an increased phenol selectivity from 3% to ~91% and almost 100% when evaluated at 30 ºC. Various Fe species, including the α-Fe(II) active site, were detected using UV-Vis spectroscopy, FTIR spectroscopy, and magnetic thermal gravimetric analysis (MTGA). Highlighting the potential of sonochemistry for preparing at mild temperatures (<100 ºC) iron-modified zeolite Y catalysts with room temperature catalytic active in water-based systems.

Modulating electronic density of active metal site via MXene intercalated by alkali ions for superior hydrogen production

Modulating electronic density of active metal site via MXene intercalated by alkali ions for superior hydrogen production

Functional significance of CYP2B6 gene rare allelic variants identified in Japanese individuals

Cytochrome P450 2B6 (CYP2B6) catalyzes the metabolism of many drugs, including efavirenz and propofol. Genetic polymorphisms in CYP2B6 alter its enzymatic activity and substantially affect its pharmacokinetics. High-frequency variants, such as CYP2B6*6, are associated with the risk of developing side effects due to reduced CYP2B6 activity. However, the impact of rare alterations on enzyme function remains unknown, and some of these variants may significantly decrease the CYP2B6 activity. Therefore, in this study, we evaluated in vitro the functional alterations in 29 missense variants of the CYP2B6 gene identified in 8,380 Japanese individuals. Wild-type CYP2B6 and 29 rare CYP2B6 variants were transiently expressed in mammalian cells. The expression levels of variant CYP2B6 proteins in the microsomal fractions extracted from 293FT cells were assessed using western blotting and reduced-carbon monoxide difference spectroscopy, and a specific peak at 450 nm was detected in the wild-type and 19 variants. Furthermore, kinetic parameters were determined by assaying the reactions with efavirenz and propofol and quantifying the metabolite concentrations. We found that 12 variants had significantly lower or abolished enzymatic activity with both the substrates. In silico three-dimensional docking and molecular-dynamics simulations suggested that these functional changes were due to conformational changes in essential regions, such as the heme-binding site and ligand channels involved in transporting substrates to the active site. These findings have implications for predicting the plasma concentrations of CYP2B6 substrates and controlling their side effects.

N-doped porous graphite with multilevel pore defects and ultra-high conductivity anchoring Pt nanoparticles for proton exchange membrane water electrolyzers

Water electrolysis for hydrogen production offers a promising solution to future energy crises and environmental challenges. Although platinum is an efficient catalyst for hydrogen evolution reactions (HERs), its high cost and stability challenges limit its widespread use. A novel platinum-based catalyst, comprising platinum nanoparticles on nitrogen-doped porous graphite (Pt-N-porous graphite), addresses these limitations. This catalyst prevents nanoparticle aggregation, provides a high specific surface area of 1308 m2 g−1, and enhances mass transfer and active site exposure. Additionally, it exhibits superior electrical conductivity compared to commercial Pt-C, enhancing charge transfer efficiency. The Pt-N-porous graphite catalyst achieves an overpotential of 99 mV at 100 mA cm−2 and maintains stable performance after 10000 cycles. Applied as a catalyst-coated membrane (CCM) in a proton exchange membrane (PEM) electrolyzer, it demonstrates excellent performance. Thus, the industrially synthesizable Pt-N-porous graphite catalyst holds great potential for large-scale energy applications.

Angular-Substituted [1,4]Thiazino[3,4-a]Isoquinolines: Biological Evaluation and In Silico Studies on DPP-IV Inhibition.

Recent studies have discovered that aryl-substituted pyrido[2,1-a]isoquinolines have the potential to be highly active DPP IV inhibitors. In previous studies, we reported a novel synthetic approach for the construction of their sulfur-containing bioisosteric [1,4]thiazino[3,4-a]isoquinolines analogues, incorporating an additional aryl substituent. The present study aims to investigate the DPP IV inhibitory activity and cytotoxicity of the synthesized molecules by in vitro assay. The geometry optimization and molecular docking of the synthesized compounds were used to determine their binding modes to the active site of DPP IV. The docking analysis revealed that the energy-minimized poses of the studied compounds are close to the most important selectivity cliffs for DPP IV inhibition, forming hydrogen bonds and hydrophobic interactions with them. These results can be considered as a preliminary step towards further structural activity modifications.

Purification, fibrinolytic activity and substrate binding of nattokinase from Bacillus subtilis: A rapid and sensitive detection for fibrinolytic activity of nattokinase

Nattokinase (NK, EC 3.4.21.62) is an alkaline serine protease secreted by B. subtilis, which has a strong fibrinolytic activity in vitro and in vivo. Here, we fermented NK using a B. subtilis strain and purified the protein, designed a peptide segment derived from fibrin as a substrate of NK. Based on fluorescence resonance energy transfer (FRET), we found that NK exhibits a high hydrolytic activity towards the peptide, with Km of 9.33 ± 1.28 μM, kcat of 0.20 ± 0.01 s−1, and kcat/Km of 21,436.23 s−1 M−1, demonstrating that the peptide is a substrate for NK. Furthermore, we investigated the binding of the substrate with NK by tryptophan fluorescence quenching, molecular docking and dynamics simulation. Fluorescence quenching showed that the substrate binds to NK mainly through hydrogen bonding and van der Waals forces, with dissociation constants KD of 13.73 and 21.24 μM at 25 and 35 °C, respectively. Molecular docking and dynamics analysis revealed that the substrate recognizes the active site of NK, and provides new information on the characteristics of the binding of the substrate with NK. Our study demonstrated a rapid and sensitive method for the quantitative measurement of fibrinolytic activity of NK based on the substrate, which is very reproducible and rapid with virtually complete results in approximately 20 min.

Redox proteomic analysis of H2O2 -treated Jurkat cells and effects of bicarbonate and knockout of peroxiredoxins 1 and 2

Oxidation of thiol proteins and redox signaling occurs in cells exposed to H2O2 but mechanisms are unclear. We used redox proteomics to seek evidence of oxidation of specific proteins either by a mechanism involving reaction of H2O2 with CO2/bicarbonate to give the more reactive peroxymonocarbonate, or via a relay involving peroxiredoxins (Prdxs). Changes in oxidation state of specific Cys-SH residues on treating Jurkat T lymphoma cells with H2O2 were measured by isotopically labeling reduced thiols and analysis by mass spectrometry. The effects of bicarbonate and of knocking out either Prdx1 or Prdx2 were examined. Approximately 14,000 Cys-peptides were detected, of which ∼ 1% underwent 2-10 fold loss in thiol content with H2O2. Those showing the most oxidation were not affected by the presence of bicarbonate or knockout of either Prdx. Consistent with previous evidence that bicarbonate potentiates inactivation of glyceraldehyde-3-phosphate dehydrogenase, the GAPDH active site Cys residues were significantly more sensitive to H2O2 when bicarbonate was present. Several other proteins were identified as promising candidates for further investigation. Although we identified some potential protein candidates for Prdx- dependent oxidation, most of the significant differences between KO and WT cells were seen in proteins for which H2O2 unexpectedly increased their CysSH content over untreated cells. We conclude that facilitation of protein oxidation by bicarbonate or Prdx-mediated relays is restricted to a small number of proteins and is insufficient to explain the majority of the oxidation of cell thiols that occurs in response to H2O2.

One-Pot Synthesis of Mixed-Phase MoVTeNbOx Catalysts for Selective Oxidation of Propane to Acrylic Acid.

One-pot hydrothermal synthesis assisted by sodium citrate and citric acid is developed to fabricate mixed-phased MoVTeNbOx catalysts with different phase compositions for selective oxidation of propane to acrylic acid. Structures of various catalysts are comprehensively characterized. Interfacial interactions occur among the M1 and M2 phases of mixed-phase MoVTeNbOx catalysts to exert synergistic effects on the selective production of acrylic acid. Various mixed-phase MoVTeNbOx catalysts exhibit the same type of active site on the M1-phase component, with a significantly enhanced ability to adsorb and activate propane. The propane conversion increases linearly with the density of surface site for propane adsorption, while the propene selectivity increases linearly with the density of surface site for propene adsorption. Among the synthesized catalysts, a MoVTeNbOx catalyst composed of 35.3 wt % M1 phase, 19.2 wt % M2 phase, and 45.7 wt % amorphous phase exhibits the largest density of active site and consequently the best catalytic performance with 38.9% propane conversion and 71.2% acrylic acid selectivity at 380 °C.

Exploration of microwave absorptivity and catalysis of CMF@Co-MoS2 for microwave-initiated depolymerization of lignin

Obtaining key platform chemicals from lignin is a sustainable refining concept of the non-petroleum route. Rapid pyrolysis is a potential refining technology, and microwave-assisted catalytic depolymerization with low energy consumption is preferred. Herein, we reported the strategy of microwave-initiated depolymerization using a self-designed dynamic vapor flow reaction system with a facilely prepared CMF@Co-MoS2-0.18 catalyst, achieving efficient conversion of lignin (Dealkalize) to monophenol. Under constant microwave irradiation (2.45 GHz), the CMF@Co-MoS2-0.18 catalyst had excellent dielectric properties (Dielectric loss factor: 0.56) and microwave heating properties (∼140 s rise to ∼ 600℃). Therefore, microwave-initiated depolymerization of lignin showed that the CMF@Co-MoS2-0.18 catalyst can improve the monophenol content (54.72 %) and phenol selectivity (35.84 %) under the synergistic effect of microwave absorptivity and catalysis. The reaction of the lignin model and kinetics analysis show that the CMF@Co-MoS2-0.18 catalyst increases the yield of monophenol by reducing depolymerization activation energy and selectively breaking the β-O-4 bond. The finite element analysis pointed out that under microwave irradiation, CMF@Co-MoS2-0.18 continuously conducted heat to lignin and the lignin vapor would have sufficient energy to collide with the catalyst active site at a high frequency, which led to an increase in the pre-exponential factor and initiated catalytic depolymerization lignin. In summary, we provide a potential pathway for the rapid conversion of lignin into key platform chemicals.

Novel Large-scale Integrated Non-Precious Metal Electrodes for Efficient and Stable Oxygen Evolution Reaction at High Current Density in 2.5 kW Anion Exchange Membrane Water Electrolysis

The utilization of non-precious catalysts to substitute precious metal catalysts on anode side under high current density conditions and with long-term resistance is crucial for the implementation of alkaline anion-exchange membrane water electrolysis (AEMWE) applications. To accommodate industrial applications, a combination of chemical solution and electrodeposition was used to construct NiFeCo layered double hydroxide and NiS nanosheet heterostructures on Ni foam (NiFeCo LDH/NiS/NF). Both experiments and theoretical calculations show that the heterogeneous structure reduces the activation energy barrier of the catalytic reaction and provides an appropriate electronic structure. This not only guarantees the exposure of the active site but also adjusts the local coordination environment and chemisorption properties. Consequently, the reduction in energy barrier affects the rate-determining step (RDS) from *O to *OOH, as well as the energy barriers of all four steps in the oxygen evolution reaction (OER). Consequently, the optimized NiFeCo LDH/NiS/NF catalyst demonstrates a low voltage of 288mV to operate 1.0Acm-2. Additionally, the catalyst was conducted with commercial Pt/C coated onto carbon paper (Pt/C/CP) for 2×2 cm2 AEMWE exhibited 1.63V cell voltag to attain 1Acm−2 and operated for 2100h from 1Acm-2 to 2.5Acm-2 (1.69V to 1.92V). Further, the catalyst was prepared scalable to 15×15 cm2 for application into a 2.5kW AEMWE device, which required only 1.77V at 60 °C to catch 1Acm-2 for 1000h, which shows extremally promising application for stable AEMWE device.

In-situ preparation of BiOBr/Bi-doped CsPbBr3 S-scheme heterojunction for efficient photocatalytic CO2 reduction

Metal halide perovskite nanocrystals have garnered significant attention in the realm of photocatalytic CO2 reduction in recent years owing to their remarkable reducing properties and high selectivity. However, the issue of low photocatalytic activity due to severe carrier recombination phenomena in perovskite has posed a significant challenge. To address this, a one-pot in-situ preparation strategy was employed to create a BiOBr/Bi-doped perovskite nanocrystal S-scheme heterojunction, resulting in a significant improvement in the CO2 photoreduction performance. The heterojunction material exhibited a CO yield of 151.56 μmolg-1h−1, with a selectivity for CO of 93.6 %. The molar ratio of the components was modulated using an in-situ water aging strategy, yielding a photocatalyst performance that exceeded that of pure BiOBr. The synthesis strategy of one-step in-situ preparation of heterojunctions overcomes the problem of insufficient bonding caused by two-step methods. And various characterizations and DFT calculations strongly support the S-scheme mechanism of the photocatalyst. The active site effectively reduces the energy barrier for the transition from *COOH to *CO intermediate, thereby significantly improving photocatalytic activity.

Structure of MltG from Mycobacterium abscessus reveals structural plasticity between composed domains.

MltG, a membrane-bound lytic transglycosylase, has roles in terminating glycan polymerization in peptidoglycan and incorporating glycan chains into the cell wall, making it significant in bacterial cell-wall biosynthesis and remodeling. This study provides the first reported MltG structure from Mycobacterium abscessus (maMltG), a superbug that has high antibiotic resistance. Our structural and biochemical analyses revealed that MltG has a flexible peptidoglycan-binding domain and exists as a monomer in solution. Further, the putative active site of maMltG was disclosed using structural analysis and sequence comparison. Overall, this study contributes to our understanding of the transglycosylation reaction of the MltG family, aiding the design of next-generation antibiotics targeting M. abscessus.

High-Valence Co Stabilized by In-Situ Growth of ZIF-67 on NiCo-LDH for Enhanced Performance in Oxygen Evolution Reaction.

The application of metal-organic frameworks (MOFs) in the electro-catalysis of heterogeneous structures is limited by the problems of low electrical conductivity and poor mechanical strength due to the complex synthesis process, although their high specific surface area and controllable structure. In this study, a method involving metal precipitation and ligand reaction is used during the electrochemical corrosion of hydroxides/oxy-hydroxides to obtain ZIF-67 in situ. The in situ growth technology not only effectively addresses the bonding strength and material conductivity challenges in the heterostructure between MOFs and the substrate but also enhances the catalyst's surface area and activity. Additionally, the exposure and protection of Co4+ by ZIF-67 contribute to the electrocatalyst's performance, demonstrating a low overpotential (η100) of 293mV, a Tafel slope of 25.8mV dec-1, and a charge transfer resistance of 3.9 Ω, with long-term robustness proven in continuous stability test exceeding 75000 s under the superhigh current density of 500mA cm-2. This work on binder-free in situ growth of MOFs not only provides relevant theoretical insights and experimental experience for cost-effective and controllable production of MOF-based catalysts but also offers ideas for the development of future electrocatalysts by exploring the exposure and protection of active site using MOFs materials.

In Silico Drug Encapsulation Using 2-Hydroxypropyl-β-CD, Tyrosine Kinase and Tyrosinase Inhibition of Dinuclear Cu(II) Carboxylate Complexes

In recent years, copper carboxylate complexes have garnered significant interest for biological applications. This study focuses on 20 Cu(II) carboxylate complexes selected from our previous research. Due to the hydrophobic nature of these complexes, the 2-hydroxypropyl-β-cyclodextrin (2HPβCD) was employed as a carrier to reduce toxicity and increase solubility for controlling drug delivery. Monte Carlo calculations were performed to confirm the interaction between the optimized structures of Cu(II) complexes and 2HPβCD, forming a host-guest system. All the structures were simulated and optimized using DFT-D calculations in Material Studio 2017. The results indicated that a neutral medium is more favorable for the adsorption of these complexes into 2HPβCD. More negative binding energy values suggested strong and energetically favorable adsorption on 2HPβCD. Complexes 4, 5, and 7 exhibited the highest interaction, making them excellent candidates for drug delivery systems. DFT-D calculations were also used to investigate the release of complexes, revealing that complexes 5, 14, and 19 were difficult to release due to their lowest energy. In contrast, complexes 8, 9, and 16 were found to be most efficient to release due to weak non-covalent interactions with 2HPβCD as we can predict from binding energy obtained by DFT-D. No specific trend was observed in the interaction of the complexes with 2HPβCD. Additionally, the effects of these complexes on c-kit tyrosine kinase and Mushroom tyrosinase were studied by molecular docking. The results demonstrated that all the complexes interacted with the active site of respective receptors through hydrophobic interactions. Complexes containing 1,10-phenanthroline and 2,2-bipyrdine were identified as having a strong, spontaneous binding ability with receptors.

Boosted Hydrogen evolution reaction based on synergistic effect of graphene, MoS2 and RuO2 ternary electrocatalyst

Hydrogen holds the promise of replacing fossil fuels and offers a sustainable pathway for energy generation. However, the large-scale production of hydrogen via the environment friendly electrocatalytic process relies heavily on the performance of electrocatalysts. In this study, we investigate the electrocatalytic performance of graphene nanosheets (GNS), molybdenum disulfide (MoS2), ruthenium dioxide (RuO2), and their composites for hydrogen evolution reaction (HER), a novel combination that has not been explored in previous literature. We synthesize the materials using Liquid Phase Exfoliation at 500 and 1000 RPMs for GNS and MoS2 and via hydrothermal methods for RuO2 nanosheets and nanoparticles, aiming to exploit synergistic effects for enhanced activity and stability. The synthesized GNS-1000/MoS2-1000/RuO2-NSs composite demonstrates promising HER results, showcasing a low overpotential of 63 mV and a reduced Tafel slope of 59 mV/dec. This improvement indicates enhanced electron transfer, improved active site dispersion, and increased surface area due to the synergistic effects, which also aids in long-term electrochemical stability. Our study underlines the potential of GNS/MoS2/RuO2 composites, particularly the GNS-1000/MoS2-1000/RuO2-NSs, in transforming hydrogen production methods and promoting efficient, sustainable energy solutions. The implications of our findings extend the boundaries of materials engineering, edging us closer to a sustainable energy future.

Synergetic Interplay of MnNi-MOF Composite with 2D MXene for Improved Supercapacitor Application

The growing demand for high-performance energy storage systems has driven the development of advanced materials to improve supercapacitor efficiency. In this study, MXene-MnNi nanocomposites are synthesized and evaluated for their potential as next-generation supercapacitor electrodes. The MnNi alloy, known for its excellent pseudocapacitive properties, is combined with MXene, a two-dimensional material recognized for its exceptional conductivity, mechanical robustness, and large surface area. The uniform dispersion of MnNi and MXene achieved through a straightforward hydrothermal method, optimizes charge transfer and enhances electrochemical performance. Electrochemical testing demonstrates that the 100 MX-MN nanocomposite exhibits a high specific capacity of 1028 C g-1 at 1 A g-1, along with excellent rate capability and long-term stability, outperforming many conventional materials. This outstanding performance is attributed to the synergistic effects of MnNi’s redox activity and MXene’s high conductivity, which together increase active site availability, accelerate ion diffusion, and maintain structural integrity. Additionally, a hybrid aqueous device is assembled using 100 MX-MN as the positive electrode and activated carbon as the negative electrode. The device delivers a maximum energy density of 87.35 W h kg-1 at a power density of 1.98 kW kg-1 and exhibits excellent capacity retention of 88 % after 10,000 charge-discharge cycles. These findings highlight the potential of MXene-MnNi nanocomposites as advanced materials for supercapacitor applications, offering a promising path for more efficient energy storage solutions in modern electronic devices.

Cobalt single-site molecular Catalyst-mediated electrochemical Hydrodechlorination for detoxification of halogenated Antibiotics: Performance, reaction pathway and mechanism

Electrocatalytic hydrodehalogenation (ECHD) offers a promising approach for detoxification of the halogenated antibiotics via hydrogenolysis of the C-X bonds (X = F, Cl or Br). Herein we demonstrated that the commercially-available cobalt phthalocyanine (CoPc) molecules with a high-loading of −[Co-N4]- moieties (Co wt%: 10.8 %) were exceptionally active for hydrogenolysis of the C-Cl bonds in florfenicol (FLO). It afforded an impressive mass activity of 6.2 gFLO h−1 g-1Co, a low Co leaching of 0.34 μg/L and a strong tolerance to interference from impurities in water, significantly surpassing reported Co/Fe/Ni-based particle and single-site counterparts. Mechanistic studies revealed that the ECHD on CoPc underwent a direct electron transfer manner rather than the atomic hydrogen (H*)-mediated indirect way, and the ECHD efficacy was affected by both the electron and proton transfer kinetics. The single-atom Co, with its d orbitals constituting the lowest unoccupied molecular orbitals of CoPc, served as the primary active site. It facilitated both electron and proton transfer, as well as the cleavage of the C-Cl bond but not the C-F bond. In a continuous-flow cell, the CoPc/C-driven ECHD reduced the antibacterial activity of a FLO-contaminated natural lake water sample by 90.8 % with an energy consumption of 0.19 kwh g-1FLO. This study highlighted great promise of the single-atom metal-bearing molecular catalyst toward the sustainable detoxification of halogenated antibiotics-contaminated water.