Chemical Modification Research Articles

Biodegradation of Pharmaceutical Pollutants Using Fungal Enzyme

Abstract: The potential of filamentous fungi and their enzymes are investigated for bioremediating pharmaceutical pollutants from wastewater thereby addressing the growing concern of emerging pollutants in water resources. Among the various microorganisms, the current abstract discusses the role of fungi in degrading antibiotics and persistent pharmaceutical compounds. These fungi allow the biodegradation via the formation of highly oxidative hydroxyl radicals. It summarizes the role of both intra (cytochrome P450) and extracellular (laccase) enzyme systems for the bioremediation of emerging clinical pollutants. It emphasizes the importance of both biosorption and biodegradation for effective pollutant removal by breaking down pharmaceuticals (e.g. antibiotics) and other hazardous compounds (e.g. endocrine-disrupting compounds, polycyclic aromatic hydrocarbons). Although the enzymes CYP450 are not directly engaged, their metabolic pathways execute mechanisms such as oxidation, the formation of reactive intermediates, and chemical modification, all of which result in the degradation of clinical pollutants. Amongst all enzyme systems in the case of laccase which oxidise a wide spectrum of substrates, converts them to free radicals. Additionally, the role of recombinant enzyme production and transgenic strains for pollutant degradation has also been elaborated. These considerations emphasise the need for ongoing research and biotechnological advancements to address the growing concern of pharmaceuticals and personal care product pollutants in water systems.

In Vivo and In Silico Molecular Modeling Studies of Newly Synthesized Pyrazoles for Their Anti-Inflammatory and Analgesic Properties

Background: Pyrazole derivatives have gained attention as potential leads for developing anti-inflammatory and analgesic drugs. Chemical modifications of pyrazole structures offer a strategy to enhance their therapeutic properties, while minimizing undesirable effects. Objective: This study aimed to synthesize novel pyrazole derivatives through chemical modifications and assess their anti-inflammatory and analgesic activities. Methods: A series of pyrazole variants were synthesized by reacting various acetophenone derivatives and diethyl oxalate in basic conditions, followed by treatment with glacial acetic acid and hydrazine hydrate. The resulting compounds were further methylated and acetylated. The synthesized compounds were characterized using elemental analysis, 1H NMR, IR, and MASS spectroscopy. The anti-inflammatory activity was evaluated using the rat paw edema technique induced by carrageenan, while the analgesic activity was assessed using the writhing reflux method. Results: Among the newly synthesized pyrazole derivatives, compounds 3b, 3e and 3g′ demonstrated significant anti-inflammatory activity with reduced ulcerogenic potential. Compounds 3d, 3b′ and 3d′ exhibited notable analgesic activity compared to the standard drug. The chemical modifications introduced in these derivatives were confirmed through spectroscopic analyses. Conclusion: The findings of this study highlight the potential of pyrazole derivatives as promising candidates for anti-inflammatory and analgesic drug development. The structural modifications performed in this research improved the therapeutic activities of the compounds, while addressing safety concerns. Further investigations are warranted to elucidate the underlying mechanisms of action and optimize the pharmacological profiles of these novel pyrazole derivatives.

Efficiency of dye adsorption of modified biochar: A comparison between chemical modification and ball milling assisted treatment

Ball milling technology was employed to increase the adsorption capacity of biochar with respect to methylene blue as a model pollutant. Comparative studies on the adsorption ability were conducted on ball-milled biochar and biochar chemically modified by oxidation and alkaline treatment, showing that ball milling produces higher performances since increases available active sites. The particle size reduction, with the consequence of an increase in the surface area was hypothesized to be responsible for the high removal efficiency. The adsorption kinetics follows a pseudo-second-order model, implying that the process is affected by chemisorption. Moreover, the Langmuir model proved that the maximum monolayer adsorption capacity increased to 185.18 mg/g. Design of experiments allowed for evaluating the pH and initial concentration values, which maximized the removal efficiency. The milled biochar was also regenerated and reused over six times, resulting in a drop of only 1 %.

A Review of Chemical Modification of Vegetable Oils and Their Applications

In order to cope with the shortage of non-renewable energy and the increasingly environmental pollution, sustainable vegetable oils, as competitive alternatives, have widely been held in the good graces of the researchers. Vegetable oils are suitable for a wide range of applications such as biofuels and biodiesel. However, the development of vegetable oils is limited due to the characteristics of unsatisfactory oxidation stability and poor cold-flow properties. Chemical modification is considered as an effective solution to enhance the performance. The research progress of the chemical modification methods and applications of vegetable oils in recent years are summarized in this review. Reducing the content of carbon–carbon double bonds and increasing the degree of saturation are the keys to improve the physicochemical properties of vegetable oils. The prospects for the development direction and challenges of vegetable oils are proposed. Future research may focus on the use of multifunctional catalysts to optimize reaction conditions or to introduce active groups with lubricating properties in epoxidation reactions and explore the combination of chemical and auxiliary methods.

Sequestration of cyanide ions from aqueous medium by physio-chemically fabricated biochar of peels of banana and grape fruit in ecofriendly way

Pakistan is an agricultural country producing plenty of fruits, like: mango, banana, apple, peaches, grapes, plums, variety of citrus fruits including lemon, grapefruit, and oranges. So far the peels of most of the fruits are usually wasted and not properly utilized anywhere. In this work, the peels of banana and grapefruit are converted into biochar by slow pyrolysis under controlled supply of air and used for sequestering cyanide ions from aqueous medium after chemical modification with ZnCl2 and sodium dodecyl sulfate (SDS). The modified biochar was characterized by various instrumental techniques, like: SEM, FTIR, TGA, and CHNS. Different parameters, like: time, temperature, pH, and dose of adsorbent affecting the adsorption of cyanide ions, onto prepared biochar were optimized and to understand the adsorption phenomenon, kinetic and thermodynamic studies were performed. Concentration of cyanide ions was estimated by employing standard ion selective electrode system and it is found that Sodium Dodecyl Sulfate treated biochar of banana peels shown more adsorption capacity, i.e.,: 17.080 mg/g as compared to all samples. Present work revealed that the biochar produced from the fruit waste has sufficient potential to eliminate trace quantities of cyanide from water, especially after treatment with sodium dodecyl sulfate.

Dynamic Electrostatic Interfacial Engineering for Block Copolymer Microparticles with Reversible Structures.

Responsive nanoparticle surfactants (NPSs) can dynamically and reversibly modulate the interfacial interactions between incompatible components, which are essential in the interfacial catalysis, corrosion, and self-assembly of block copolymers (BCPs). However, NPSs with stimuli-responsive behavior often involve tedious chemical synthesis and surface modifications. Herein, we propose a strategy to in situ construct a kind of dynamic and reversible NPSs by the interfacial electrostatic interaction between the negatively charged nanoparticles (NPs) and the positively charged homopolymers. The NPSs assembled at the oil/water interface reduce the interfacial tension and direct the confined assembly of BCP. Meanwhile, the dynamic NPSs can be disassembled by increasing the pH value or introducing competitive electrostatic attractions, which can dynamically and reversibly change the interfacial properties as well as the alignment of polymer chains, enabling BCP microparticles with reversibly switchable lamellar and cylindrical structures. Furthermore, by the introduction of aggregation-induced emission luminogens as tails to the NPSs, the reversible transformation of BCP microparticles can be visualized by fluorescence emission, which is dependent on the nanostructures of microparticles. This work establishes a concept for dynamically manipulating interfacial interactions and reversibly switching BCP microparticles without time-consuming NPS synthesis, showing promising applications in the fabrication of smart materials with switchable structures and properties.

Genetic regulation of m6A RNA methylation and its contribution in human complex diseases.

N6-methyladenosine (m6A) has been established as the most prevalent chemical modification in message RNA (mRNA), playing an essential role in determining the fate of RNA molecules. Dysregulation of m6A has been revealed to lead to abnormal physiological conditions and cause various types of human diseases. Recent studies have delineated the genetic regulatory maps for m6A methylation by mapping the quantitative trait loci of m6A (m6A-QTLs), thereby building up the regulatory circuits linking genetic variants, m6A, and human complex traits. Here, we review the recent discoveries concerning the genetic regulatory maps of m6A, describing the methodological and technical details of m6A-QTL identification, and introducing the key findings of the cis- and trans-acting drivers of m6A. We further delve into the tissue- and ethnicity-specificity of m6A-QTL, the association with other molecular phenotypes in light of genetic regulation, the regulators underlying m6A genetics, and importantly, the functional roles of m6A in mediating human complex diseases. Lastly, we discuss potential research avenues that can accelerate the translation of m6A genetics studies toward the development of therapies for human genetic diseases.

The Applications of Nanopore Sequencing Technology in Animal and Human Virus Research

In recent years, an increasing number of viruses have triggered outbreaks that pose a severe threat to both human and animal life, as well as caused substantial economic losses. It is crucial to understand the genomic structure and epidemiology of these viruses to guide effective clinical prevention and treatment strategies. Nanopore sequencing, a third-generation sequencing technology, has been widely used in genomic research since 2014. This technology offers several advantages over traditional methods and next-generation sequencing (NGS), such as the ability to generate ultra-long reads, high efficiency, real-time monitoring and analysis, portability, and the ability to directly sequence RNA or DNA molecules. As a result, it exhibits excellent applicability and flexibility in virus research, including viral detection and surveillance, genome assembly, the discovery of new variants and novel viruses, and the identification of chemical modifications. In this paper, we provide a comprehensive review of the development, principles, advantages, and applications of nanopore sequencing technology in animal and human virus research, aiming to offer fresh perspectives for future studies in this field.

The Functions of N-methyladenosine (m6A) Modification on HIV-1 mRNA.

In recent years, there has been a growing interest in the study of RNA modifications, with some researchers focusing specifically on the connection between these modifications and viruses, as well as the impact they have on viral mRNA and its functionality. The most common type of RNA chemical modification is m6A, which involves the addition of a methyl group covalently to the N6 position of adenosine. It is a widely observed and evolutionarily conserved RNA modification. The regulation of m6A modification primarily involves methyltransferases (writers) and demethylases (erasers) and is mediated by m6A-binding proteins (readers). In HIV-1, m6A sites are predominantly located in the 5' untranslated region (5'UTR) and 3' untranslated region (3'UTR). Additionally, m6A modifications are also present in the RRE RNA of HIV-1. This review provides a detailed account of the effects of these m6A modifications on HIV-1 functionality.

Modified Starch from Mango Pickling Industry Waste: Comparison of Physical and Chemical Modification

AbstractThis study focuses on extracting, modifying, and characterizing mango kernel starch (MKS) from raw pickling mango waste. Two physical and two chemical techniques: heat‐moisture treatment (HMT), ultrasound (US), sodium tri‐metaphosphate (STMP), and octenyl succinic anhydride (OSA) are employed for modification. HMT and STMP reduce swelling power and solubility due to structural changes, while US and OSA treatments enhance these properties by disrupting intermolecular bonds and weakening hydrogen bonds within the starch granules. Fourier transform infrared spectroscopy (FTIR) spectra demonstrate changes in absorption peaks, intensities, and positions in the modified starches, confirming successful structural alterations. X‐ray diffraction patterns indicate A‐type and C‐type patterns for native and modified starches, revealing alterations in crystallinity driven by modification techniques. Water binding capacity (WBC) generally increased in modified starches compared to native starch. Changes in amylose content and pasting properties are observed. Modified starches display higher thermal stability. Furthermore, starch retrogradation is affected by these modifications, with STMP and OSA modification treatments slowing it down, HMT and US treatments promoting it. Results demonstrate that native and modified MKS can be versatile ingredients for various industries. Chemically modified MKS excelled in crystallinity and hindered retrogradation, making it beneficial for edible coatings and baked goods. Physically modified MKS demonstrated accelerated retrogradation and suited for extruded snacks.

Chemical Modification of Birch Bark (Betula L.) for the Improved Bioprocessing of Cadmium(II), Chromium(VI), and Manganese(II) from Aqueous Solutions

This study aimed to assess the sorption capacity of a natural sorbent, specifically birch bark (BB), and its modification using chemical reagents, including nitric and hydrochloric acid, sodium hydroxide, and chloride. The objective of the chemical modification was to enhance the sorption capacity of the heavy metals cadmium(II), chromium(VI), and manganese(II). The most effective modification for adsorbing cadmium and manganese from aqueous solutions was achieved by treating the sorbent with a 0.1 M sodium hydroxide solution (BBNa). Conversely, in the case of chromium, each modification adversely affected its adsorption by the sorbent. Concentrations of the solutions were analyzed using atomic absorption spectrometry at appropriate time intervals. The adsorption process was described using Langmuir, Freundlich, and Temkin isotherms. The Freundlich isotherm provided the best fit for cadmium and chromium (R2 = 0.988 and 0.986, respectively), while the Langmuir isotherm was most suitable for manganese (R2 = 0.996). The sorption capacity varied for each metal ion: Cd (II)—33.13 mg/g, Cr (VI)—35.98 mg/g, and Mn (II)—24 mg/g for the highest concentration tested. This study employed pseudo–first-rate order, pseudo–second-rate order model kinetics, and the Weber–Morris model to examine the adsorption kinetics. The pseudo–second-rate order kinetics demonstrated the best fit (R2 > 0.94) for each heavy metal, which underlines the process’s chemical nature.

Properties of biopolymer blends based on Rugulopteryx okamurae and hydrophobic polycaprolactone (PCL) and hydrophilic acylated soy protein isolated (SPIa).

The present study explored the utilization of Rugulopteryx okamurae (RO), an invasive brown seaweed, as a renewable raw material for plastic materials based on biopolymer blends. The goal of this study was to improve the previously observed poor mechanical properties of materials based on single biopolymer RO. To enhance these properties, two polymers with distinct hydrophobicities were incorporated into the formulation of different blends: hydrophobic polycaprolactone (PCL) and hydrophilic acylated soy protein isolate (SPIa). SPIa was derived from soy protein through a chemical modification process, introducing hydrophilic carboxyl groups. The addition of PCL significantly strengthened the blend, increasing the storage modulus (E'1Hz) from ~ 110 to ~ 250MPa. Conversely, SPIa incorporation resulted in softening, with E' values around 40MPa. Both additives enhanced deformability proportionally to their concentrations, with SPIa exhibiting notably higher deformability, reaching a maximum deformation of ~ 23% for a RO/SPIa ratio of 25/75. In summary, the study demonstrates the feasibility of producing environmentally friendly blend materials based on RO, tailored for specific applications by incorporating suitable additives into the formulation. Therefore, PCL is recommended for applications susceptible to moisture effects, such as packaging, while SPIa is suggested for highly absorbent applications such as personal care or horticulture.

Purine, nucleoside, and DNA nitration by peroxynitrite: Mechanistic considerations

Nitro-oxidative stress affects DNA, leading to special chemical modifications of nucleobases and deoxyribose, impacting DNA integrity and stability. Because of the importance of the topic, the state of the knowledge on purine, nucleoside, and DNA nitration by the reactive nitrogen species peroxynitrite was reviewed. Following a description of the chemical and physicochemical characteristics of purines and peroxynitrite, purine nitro-oxidation and its products, the reaction mechanisms, and the recently reported kinetic behavior of 8-NitroGua formation are discussed. Moreover, novel computational studies report structural and conformational DNA changes resulting from the formation of guanine nitration products. Given the relevance of the subject, surprisingly few publications deal with this topic, even considering the past five years.

The molecular interaction of six single-stranded DNA aptamers to cardiac troponin I revealed by docking and molecular dynamics simulation.

Cardiac troponin I (cTnI) is a cardiac biomarker for diagnosing ischemic heart disease and acute myocardial infarction. Current biochemical assays use antibodies (Abs) due to their high specificity and sensitivity. However, there are some limitations, such as the high-cost production of Abs due to complex instruments, reagents, and steps; the variability of Abs quality from batch to batch; the low stability at high temperatures; and the difficulty of chemical modification. Aptamer overcomes the limitations of antibodies, such as relatively lower cost, high reproducibility, high stability, and ease of being chemically modified. Aptamers are three-dimensional architectures of single-stranded RNA or DNA that bind to targets such as proteins. Six aptamers (Tro1-Tro6) with higher binding affinity than an antibody have been identified, but the molecular interaction has not been studied. In this study, six DNA aptamers were modeled and docked to cTnI protein. Molecular docking revealed that the interaction between all aptamer and cTnI happened in the similar cTnI region. The interaction between aptamer and cTnI involved hydrophobic interaction, hydrogen bonds, π-cation interactions, π-stack interactions, and salt-bridge formation. The calculated binding energy of all complexes was negative, which means that the complex formation was thermodynamically favorable. The electrostatic energy term was the main driving force of the interaction between all aptamer and cTnI. This study could be used to predict the behavior of further modified aptamer to improve aptamer performance.

Collagen-Mesenchymal Stem Cell Microspheres Embedded in Hyaluronic Acid Solutions as Biphasic Stem Niche Delivery Systems for Pulmonary Differentiation.

Cell therapy has the potential to become a feasible solution for several diseases, such as those related to the lungs and airways, considering the more beneficial intratracheal administration route. However, in lung diseases, an impaired pulmonary extracellular matrix (ECM) precludes injury resolution with a faulty engraftment of mesenchymal stem cells (MSCs) at the lung level. Furthermore, a shielding strategy to avoid cell damage as well as cell loss due to backflow through the injection path is required. Here, an approach to deliver cells encapsulated in a biomimetic stem niche is used, in which the interplay between cells and physiological lung ECM constituents, such as collagen and hyaluronic acid (HA), can occur. To this aim, a biphasic delivery system based on MSCs encapsulated in collagen microspheres (mCOLLs) without chemical modification and embedded in an injectable HA solution has been developed. Such biphasic delivery systems can both increase the mucoadhesive properties at the site of interest and improve cell viability and pulmonary differentiation. Rheological results showed a similar viscosity at high shear rates compared to the MSC suspension used in intratracheal administration. The size of the mCOLLs can be controlled, resulting in a lower value of 200 μm, suitable for delivery in alveolar sacs. Biological results showed that mCOLLs maintained good cell viability, and when they were suspended in lung medium implemented with low molecular weight HA, the differentiation ability of the MSCs was further enhanced compared to their differentiation ability in only lung medium. Overall, the results showed that this strategy has the potential to improve the delivery and viability of MSCs, along with their differentiation ability, in the pulmonary lineage.

Chemical and Enzyme-Mediated Chemical Reactionsfor Studying Nucleic Acids and Their Modifications.

Nucleic acids are genetic information-carrying molecules inside cells. Apart from basic nucleotide building blocks, there exist various naturally occurring chemical modifications on nucleobase and ribose moieties, which greatly increase the encoding complexity of nuclei acids, contribute to the alteration of nucleic acid structures, and play versatile regulation roles in gene expression. To study the functions of certain nucleic acids in various biological contexts, robust tools to specifically label and identify these macromolecules and their modifications, and to illuminate their structures are highly necessary. In this review, we summarize recent technique advances of using chemical and enzyme-mediated chemical reactions to study nucleic acids and their modifications and structures. By highlighting the chemical principles of these techniques, we aimtopresent a perspectiveon the advancement of the field as well as to offerinsights intodeveloping specific chemical reactions and precise enzyme catalysis utilized for nucleic acids and their modifications.

Inhibition of endocytic uptake of severe acute respiratory syndrome coronavirus 2 and endo-lysosomal acidification by diphenoxylate.

Cell culture-based screening of a chemical library identified diphenoxylate as an antiviral agent against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The observed 50% effective concentrations ranged between 1.4 and 4.9 µM against the original wild-type strain and its variants. Time-of-addition experiments indicated that diphenoxylate is an entry blocker targeting a host factor involved in viral infection. Fluorescence microscopic analysis visualized that diphenoxylate prevented SARS-CoV-2 particles from penetrating the cell membrane and also impaired endo-lysosomal acidification. Diphenoxylate exhibited a synergistic inhibitory effect on SARS-CoV-2 infection in human lung epithelial Calu-3 cells when combined with a transmembrane serine protease 2 (TMPRSS2) inhibitor, nafamostat. This synergy suggested that efficient antiviral activity is achieved by blocking both TMPRSS2-mediated early and endosome-mediated late SARS-CoV-2 entry pathways. The antiviral efficacy of diphenoxylate against SARS-CoV-2 was reproducible in a human tonsil organoids system. In a transgenic mouse model expressing the obligate SARS-CoV-2 receptor, human angiotensin-converting enzyme 2, intranasal administration of diphenoxylate (10 mg/kg/day) significantly reduced the viral RNA copy number in the lungs by 70% on day 3. This study underscores that diphenoxylate represents a promising core scaffold, warranting further exploration for chemical modifications aimed at developing a new class of clinically effective antiviral drugs against SARS-CoV-2.

Synthesis and anti-inflammatory activities of two new N-acetyl glucosamine derivatives

N-acetyl glucosamine (NAG) is a natural amino sugar found in various human tissues with previously described anti-inflammatory effects. Various chemical modifications of NAG have been made to promote its biomedical applications. In this study, we synthesized two bi-deoxygenated NAG, BNAG1 and BNAG2 and investigated their anti-inflammatory properties, using an in vivo and in vitro inflammation mouse model induced by lipopolysaccharide (LPS). Among the parent molecule NAG, BNAG1 and BNAG2, BNAG1 showed the highest inhibition against serum levels of IL-6 and TNF α and the leukocyte migration to lungs and peritoneal cavity in LPS challenged mice, as well as IL-6 and TNF α production in LPS-stimulated primary peritoneal macrophages. BNAG2 displayed an anti-inflammatory effect which was comparable to NAG. These findings implied potential application of these novel NAG derivatives, especially BNAG1, in treatment of certain inflammation-related diseases.

Magnetically modified-mitoxantrone mesoporous organosilica drugs: an emergent multimodal nanochemotherapy for breast cancer

BackgroundChemotherapy, the mainstay treatment for metastatic cancer, presents serious side effects due to off-target exposure. In addition to the negative impact on patients’ quality of life, side effects limit the dose that can be administered and thus the efficacy of the drug. Encapsulation of chemotherapeutic drugs in nanocarriers is a promising strategy to mitigate these issues. However, avoiding premature drug release from the nanocarriers and selectively targeting the tumour remains a challenge.ResultsIn this study, we present a pioneering method for drug integration into nanoparticles known as mesoporous organosilica drugs (MODs), a distinctive variant of periodic mesoporous organosilica nanoparticles (PMOs) in which the drug is an inherent component of the silica nanoparticle structure. This groundbreaking approach involves the chemical modification of drugs to produce bis-organosilane prodrugs, which act as silica precursors for MOD synthesis. Mitoxantrone (MTO), a drug used to treat metastatic breast cancer, was selected for the development of MTO@MOD nanomedicines, which demonstrated a significant reduction in breast cancer cell viability. Several MODs with different amounts of MTO were synthesised and found to be efficient nanoplatforms for the sustained delivery of MTO after biodegradation. In addition, Fe3O4 NPs were incorporated into the MODs to generate magnetic MODs to actively target the tumour and further enhance drug efficacy. Importantly, magnetic MTO@MODs underwent a Fenton reaction, which increased cancer cell death twofold compared to non-magnetic MODs.ConclusionsA new PMO-based material, MOD nanomedicines, was synthesised using the chemotherapeutic drug MTO as a silica precursor. MTO@MOD nanomedicines demonstrated their efficacy in significantly reducing the viability of breast cancer cells. In addition, we incorporated Fe3O4 into MODs to generate magnetic MODs for active tumour targeting and enhanced drug efficacy by ROS generation. These findings pave the way for the designing of silica-based multitherapeutic nanomedicines for cancer treatment with improved drug delivery, reduced side effects and enhanced efficacy.

Role of atomic substitution in first coordination shell of Fe–N–C single atom catalyst towards oxygen reduction reaction: A theoretical study

Due to structural complexity and limited understanding, precise chemical modification in atomically dispersed metal-nitrogen-carbon (M-N-C) catalysts requires theoretical calculations to optimize electronic, geometric, and catalytic performance and experimental implementation of learned lessons to improve their catalytic performance towards oxygen reduction reaction (ORR). Herein, we conducted a density functional theory (DFT) investigation to understand the effect of the replacement of O-atoms in Fe–O4–C by N-atoms in the first coordination shell of Fe-site (constructing five models: Fe–O4–C, Fe–O3N–C, Fe–N2O2–C, Fe–ON3–C and Fe–N4–C) towards ORR. The geometric and electronic calculations of these models suggested that replacing all the O-atoms with N-atoms around the Fe-site would make it more conducive to the adsorption of ORR intermediates. Moreover, the Gibbs free energy calculations demonstrated that the Fe-site becomes the active centre for 2e− ORR, leading to the dominant generation of H2O2 when Fe is coordinated with four O-atoms (Fe–O4–C). Whereas in compounds like Fe–N2O2–C, Fe–O3N–C, and Fe–N4–C, where the Fe-site displays 4e− ORR, form water is a preferred product if the O-atoms are replaced by N-atoms, as in Fe–N2O2–C, Fe–O3N–C, and Fe–N4–C. This study would provide a fundamental understanding of how atomically dispersed M-N-C catalysts' electrocatalytic activity is affected by changes in the coordination environment of metal sites.