Reductive Methylation Research Articles

Midseason draining of paddy water suppresses microbial arsenic methylation in soil and alleviates rice straighthead disease

Arsenic (As) methylation is an important component of As biogeochemical cycle. Microbial As methylation is enhanced under anoxic conditions in paddy soil, producing dimethylarsenate (DMA) which can cause physiological straighthead disease in rice. We conducted field experiments at two sites to test the effect of midseason draining of paddy water on microbial As methylation and the incidence of straighthead disease. Compared to continuous flooding, midseason draining increased soil Eh, decreased the abundances of microbial genes for arsenate reduction (arsC and arrA) and arsenite methylation (arsM), and lowered the concentrations of both inorganic As and DMA in soil porewater. Draining shifted microbial composition, resulting in decreases in the relative abundance of 17–132 amplicon sequence variants. Draining decreased the accumulation of DMA in rice husk and of inorganic As and DMA in rice grain, decreased the incidence of straighthead disease, and increased grain yield by 20–45 %. Further experiments were conducted at eight field sites to assess the effect of midseason draining in a split field design. Draining decreased husk DMA concentration by 40–65 % and increased grain yield by 25–209 %. This study demonstrates that midseason draining can effectively suppress microbial As methylation and alleviate rice straighthead disease, benefiting both grain yield and safety.

Lysine methylation: A strategy to improve in-cell NMR spectroscopy of proteins

BackgroundIn-cell NMR is a valuable technique for investigating protein structure and function in cellular environments. However, challenges arise due to highly crowded cellular environment, where nonspecific interactions between the target protein and other cellular components can lead to signals broadening or disappearance in NMR spectra. ResultsWe implemented chemical reduction methylation to selectively modify lysine residues on protein surfaces aiming to weaken charge interactions and recover obscured NMR signals. This method was tested on six proteins varying in molecular size and lysine content. While methylation did not disrupt the protein's native conformation, it successful restored some previously obscured in-cell NMR signals, particularly for proteins with high isoelectric points that decreased post-methylation. SignificanceThis study affirms lysine methylation as a feasible approach to enhance the sensitivity of in-cell NMR spectra for protein studies. By mitigating signal loss due to nonspecific interactions, this method expands the utility of in-cell NMR for investigating proteins in their natural cellular environment, potentially leading to more accurate structural and functional insights.

Nickel-Catalyzed Reductive Methylation of Secondary Mesylates with Methyl Tosylate

Nickel-Catalyzed Reductive Methylation of Secondary Mesylates with Methyl Tosylate

Cross‐selective Deoxygenative Coupling of Aliphatic Alcohols: Installation of Methyl Groups including Isotopic Labels by Nickel Catalysis

AbstractNickel‐catalyzed cross‐electrophile coupling reactions of two aliphatic alcohol derivatives remain a challenge. Herein, we report a nickel‐catalyzed reductive methylation reaction of aliphatic mesylates with methyl tosylate. This reaction provides straightforward access to compounds bearing aliphatic methyl groups from alkyl alcohol derivatives. Isotopically labelled substrates and reagents can be employed in the reaction to provide perdeuterated and 13C‐labelled products. This transformation can be achieved by employing stoichiometric Mn reductant or electrochemically. Additionally, mechanistic experiments show that alkyl iodides are key intermediates in the transformation which undergo a stereoablative reaction via radical intermediates.

Cross-selective Deoxygenative Coupling of Aliphatic Alcohols: Installation of Methyl Groups including Isotopic Labels by Nickel Catalysis.

Nickel-catalyzed cross-electrophile coupling reactions of two aliphatic alcohol derivatives remain a challenge. Herein, we report a nickel-catalyzed reductive methylation reaction of aliphatic mesylates with methyl tosylate. This reaction provides straightforward access to compounds bearing aliphatic methyl groups from alkyl alcohol derivatives. Isotopically labelled substrates and reagents can be employed in the reaction to provide perdeuterated and 13C-labelled products. This transformation can be achieved by employing stoichiometric Mn reductant or electrochemically. Additionally, mechanistic experiments show that alkyl iodides are key intermediates in the transformation which undergo a stereoablative reaction via radical intermediates.

First row transition metal catalyzed small molecule activation for sustainable organic transformation: Synergy between nature and chemist’s toolbox

In this review we have tried to present our view on the synergy between nature and chemists’ toolbox toward discovery of new reactions. Toward the search for new chemical reactivity, not only chemists learn from nature but nature’s evolutionary pathway perhaps has footprints of chemist’s innovative discoveries and beyond. Taking CO2 as a representative example of an abundant energy rich small molecule resource, we have tried to explore the probable strategies of activating the molecule using first row transition metal catalysts for sustainable organic transformation. Our study has tried to unfold nature’s strategy of CO2 activation and recycling which eventually motivated chemists toward discovery of new chemical reactions and further inspired to expand their toolbox for discovering new reactivities beyond the natural enzyme repertoire. The basic chemical CO2 activation strategy which have been focused in this review are using CO2 as a C1 building block in organic synthesis; catalytic reductive formylation and methylation of amines; carboxylation reactions via CO2 insertion. Additionally, carbonylation reactions with CO2 as CO surrogate has also been discussed. The focus of this review is to correlate nature’s chemical strategies with chemist’s approach to understand the concept involved in the chemical activation and utilization process of CO2 for unlocking new reaction paradigm, both for chemists’ and for nature’s catalytic repertoire.

One-pot two-step reduction of CO2 with amines and NaBH4 to N-substituted compounds at atmospheric pressure

In this paper, the reductive N-formylation, methylation and cyclization of amines with carbon dioxide (CO2) using sodium borohydride (NaBH4) at atmospheric pressure, via a one-pot two-step procedure, is presented. Formato borohydride (NaBHn(OCHO)4-n), in situ formed in the reaction of NaBH4 with CO2, as an efficient agent to promote the corresponding reaction, and the mechanism is proposed on the basis of experiments. This wide substrate scope and simple operational methodology also facilitates the effective synthesis of formamides, methylamines, benzimidazoles and other heterocyclic compounds avoiding high temperature and pressure.

Understanding the pharmacokinetic journey of Fc-fusion protein, rhIL-7-hyFc using complementary approach of two analytical methods, accelerator mass spectrometry and ELISA.

Antibody-based therapeutics (ABTs), including monoclonal/polyclonal antibodies and fragment crystallizable region (Fc)-fusion proteins, are increasingly used in disease treatment, driving the global market growth. Understanding the pharmacokinetic (PK) properties of ABTs is crucial for their clinical effectiveness. This study investigated the PK profile and tissue distribution of efineptakin alfa, a long-acting recombinant human interleukin-7 (rhIL-7-hyFc), using enzyme-linked immunosorbent assay (ELISA) and accelerator mass spectrometry (AMS). Totally, four rats were injected intramuscularly with 1mg/kg of rhIL-7-hyFc containing 14C-rhIL-7-hyFc, which was prepared via reductive methylation. Serum total radioactivity (TRA) and serum rhIL-7-hyFc concentrations were quantified using AMS and ELISA, respectively. The TRA concentrations in organs were determined by AMS. Serum TRA peaked at 10hours with a terminal half-life of 40hours. The rhIL-7-hyFc exhibited a mean peak concentration at around 17hours and a rapid elimination with a half-life of 12.3hours. Peak concentration and area under the curve of TRA were higher than those of rhIL-7-hyFc. Tissue distribution analysis showed an elevated TRA concentrations in lymph nodes, kidneys, and spleen, indicating rhIL-7-hyFc's affinity for these organs. The study also simulated the positions of 14C labeling in rhIL-7-hyFc, identifying specific residues in the fragment of rhIL-7 portion, and provided the explanation of distinct analytes targeted by each method. Combining ELISA and AMS provided advantages by offering sensitivity and specificity for quantification as well as enabling the identification of analyte forms. The integrated use of ELISA and AMS offers valuable insights for the development and optimization of ABT.

Quantitative Peptidomics Using Reductive Methylation of Amines.

A number of different approaches have been used for quantitative peptidomics. In this protocol, we describe the method in which peptides are reacted with formaldehyde and sodium cyanoborohydride, which converts primary and secondary amines into tertiary amines. By using different combinations of regular reagents, deuterated reagents (2H), and reagents containing deuterium and 13C, it is possible to produce five isotopically distinct forms of the methylated peptides, which can be quantified by mass spectrometry. Peptides with free N-termini that are primary amines incorporate two methyl groups using this procedure, which differ by 2Da for each of the five isotopic combinations. Peptides that contain unmodified lysine residues incorporate additional pairs of methyl groups, leading to larger mass differences between isotopic forms. The reagents are commercially available, relatively inexpensive, and chemically stable.

Dissolved organic matter mediated dark- and photo-aging processes of Hg(II): Critical impacts of binding sites and sulfidation on Hg(II) abiotic reduction and microbial methylation

Dissolved organic matter (DOM)-mediated divalent mercury (Hg(II)) aging kinetics play a crucial role in controlling Hg(II) transformation and bioavailability in natural aquatic environments. However, the differential environmental behaviors of new and aged Hg(II) in a same reaction system remains unknown. In this study, multi-isotope tracing was used to investigate the impacts of binding site and sulfidation during DOM-mediated Hg(II) aging processes on Hg(II) reduction and microbial methylation in the same reaction system. Stepwise reduction approach and liquid chromatography-inductively coupled plasma mass spectrometry (LC-ICP-MS) demonstrate that DOM-mediated dark aging processes are mainly driven by the rearrangement of DOM binding sites with Hg(II), but not the formation of mercury sulfide nanoparticles (HgSNP). The abundant but weaker RO/N (carboxyl and amino) Hg(II)-binding sites are replaced with stronger RSH (thiol) moieties towards Hg(II) binding with aging, resulting in a decrease in Hg(II) reduction. In contrast, besides reduction, DOM-mediated Hg(II) photoaging induces the formation of HgSNP, as revealed by LC-ICP-MS, which in turn decreases the microbial methylation potential of Hg(II). These findings help better understand and predict the kinetic characteristics of Hg(II) reactivity and its influence on Hg cycle within natural aquatic environments.

Selective reductive α-methylation of chalcone derivatives using methanol

An electron rich, new co-operative iridium(III) complex catalysed tandem conversion of α,β-unsaturated ketones to the α-methylated ketones is described using methanol. Employing this chemoselective protocol, reductive methylation of a wide range of chalcone derivatives, variety of aryl and aliphatic fragment containing α,β-unsaturated ketones along with few biologically important molecules were effectively carried out. Several control experiments, kinetic studies, Hammett study, and DFT calculations were accomplished to comprehend this co-operative iridium(III) catalysed tandem reductive methylation of α,β-unsaturated ketones.

Microbial community composition in the rhizosphere of Pteris vittata and its effects on arsenic phytoremediation under a natural arsenic contamination gradient.

Arsenic contamination causes numerous health problems for humans and wildlife via bioaccumulation in the food chain. Phytoremediation of arsenic-contaminated soils with the model arsenic hyperaccumulator Pteris vittata provides a promising way to reduce the risk, in which the growth and arsenic absorption ability of plants and the biotransformation of soil arsenic may be greatly affected by rhizosphere microorganisms. However, the microbial community composition in the rhizosphere of P. vittata and its functional role in arsenic phytoremediation are still poorly understood. To bridge this knowledge gap, we carried out a field investigation and pot experiment to explore the composition and functional implications of microbial communities in the rhizosphere of four P. vittata populations with a natural arsenic contamination gradient. Arsenic pollution significantly reduced bacterial and fungal diversity in the rhizosphere of P. vittata (p < 0.05) and played an important role in shaping the microbial community structure. The suitability of soil microbes for the growth of P. vittata gradually decreased following increased soil arsenic levels, as indicated by the increased abundance of pathogenic fungi and parasitic bacteria and the decrease in symbiotic fungi. The analysis of arsenic-related functional gene abundance with AsChip revealed the gradual enrichment of the microbial genes involved in As(III) oxidation, As(V) reduction, and arsenic methylation and demethylation in the rhizosphere of P. vittata following increased arsenic levels (p < 0.05). The regulation of indigenous soil microbes through the field application of fungicide, but not bactericide, significantly reduced the remediation efficiency of P. vittata grown under an arsenic contamination gradient, indicating the important role of indigenous fungal groups in the remediation of arsenic-contaminated soil. This study has important implications for the functional role and application prospects of soil microorganisms in the phytoremediation of arsenic-polluted soil.

One-Pot Reductive Methylation of Nitro- and Amino-Substituted (Hetero)Aromatics with DMSO/HCOOH: Concise Synthesis of Fluorescent Dimethylamino-Functionalized Bibenzothiazole Ligands with Tunable Emission Color upon Complexation.

One-pot reductive N,N-dimethylation of suitable nitro- and amino-substituted (hetero)arenes can be achieved using a DMSO/HCOOH/Et3N system acting as a low-cost but efficient reducing and methylating agent. The transformation of heteroaryl-amines can be accelerated by using dimethyl sulfoxide/oxalyl chloride or chloromethyl methyl sulfide as the source of active CH3SCH2+ species, while the exclusion of HCOOH in the initial stage of the reaction allows avoiding N-formamides as resting intermediates. The developed procedures are applicable in multigram-scale synthesis, and because of the lower electrophilicity of CH3SCH2+, they also work in pathological cases, where common methylating agents provide N,N-dimethylated products in no yield or inferior yields due to concomitant side reactions. The method is particularly useful in one-pot reductive transformation of 2-H-nitrobenzazoles to corresponding N,N-dimethylamino-substituted heteroarenes. These, upon Cu(II)-catalyzed oxidative homocoupling, afford 2,2'-bibenzazoles substituted with dimethylamino groups as charge-transfer N^N ligands with intensive absorption/emission in the visible region. The fluorescence of NMe2-functionalized bibenzothiazoles remains intensive even upon complexation with ZnCl2, while emission maxima are bathochromically shifted from the green/yellow to orange/red spectral region, making these small-molecule fluorophores, exhibiting large emission quantum yields and Stokes shifts, an attractive platform for the construction of various functional dyes and light-harvesting materials with tunable emission color upon complexation.

Electrochemical Reductive N-Methylation with CO2 Enabled by a Molecular Catalyst

The development of benign methylation reactions utilizing CO2 as a one-carbon building block would enable a more sustainable chemical industry. Electrochemical CO2 reduction has been extensively studied, but its application for reductive methylation reactions remains out of the scope of current electrocatalysis. Here, we report the first electrochemical reductive N-methylation reaction with CO2 and demonstrate its compatibility with amines, hydroxylamines, and hydrazine. Catalyzed by cobalt phthalocyanine molecules supported on carbon nanotubes, the N-methylation reaction proceeds in aqueous media via the chemical condensation of an electrophilic carbon intermediate, proposed to be adsorbed or near-electrode formaldehyde formed from the four-electron reduction of CO2, with nucleophilic nitrogenous reactants and subsequent reduction. By comparing various amines, we discover that the nucleophilicity of the amine reactant is a descriptor for the C-N coupling efficacy. We extend the scope of the reaction to be compatible with cheap and abundant nitro-compounds by developing a cascade reduction process in which CO2 and nitro-compounds are reduced concurrently to yield N-methylamines with high monomethylation selectivity via the overall transfer of 12 electrons and 12 protons.

Sample Preparation and Relative Quantitation using Reductive Methylation of Amines for Peptidomics Studies

Sample Preparation and Relative Quantitation using Reductive Methylation of Amines for Peptidomics Studies

Sample Preparation and Relative Quantitation using Reductive Methylation of Amines for Peptidomics Studies.

Peptidomics can be defined as the qualitative and quantitative analysis of peptides in a biological sample. Its main applications include identifying the peptide biomarkers of disease or environmental stress, identifying neuropeptides, hormones, and bioactive intracellular peptides, discovering antimicrobial and nutraceutical peptides from protein hydrolysates, and can be used in studies to understand the proteolytic processes. The recent advance in sample preparation, separation methods, mass spectrometry techniques, and computational tools related to protein sequencing has contributed to the increase of the identified peptides number and peptidomes characterized. Peptidomic studies frequently analyze peptides that are naturally generated in cells. Here, a sample preparation protocol based on heat-inactivation is described, which eliminates protease activity, and extraction with mild conditions, so there is no peptide bonds cleavage. In addition, the relative quantitation of peptides using stable isotope labeling by reductive methylation of amines is also shown. This labeling method has some advantages as the reagents are commercially available, inexpensive compared to others, chemically stable, and allows the analysis of up to five samples in a single LC-MS run.

Reductive N-methylation of alkanolamines with paraformaldehyde in the presence of cobalt catalysts

Herein a simple method for N-methylation of ethanolamines with paraformaldehyde as a dual methylation and reducing agent has been investigated. Two new cobalt complexes, Co[Methyl 6-(morpoline methyl) picolinate]2Cl2 and Co(2,6 bis morpholine methyl pyridine)Cl2 were prepared and applied as homogeneous catalyst in reductive methylation reaction. All experiments have progressed with excellent conversion and selectivity toward N,N dimethyl ethanolamine. On the other hand, Pyrolysis of prepared cobalt complexes on activated carbon at 550 °C leads to the formation of cobalt oxide nano-particles. These heterogeneous catalysts were shown similar activities in this reaction. Single-crystal X-ray crystallography, Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) were used for characterization of prepared catalysts.

Deaminative Reductive Methylation of Alkylpyridinium Salts.

Methyl groups can imbue valuable properties in organic molecules, often leading to enhanced bioactivity. To enable efficient installation of methyl groups on simple building blocks and in late-stage functionalization, a nickel-catalyzed reductive coupling of secondary Katritzky alkylpyridinium salts with methyl iodide was developed. When coupled with formation of the pyridinium salt from an alkyl amine, this method allows amino groups to be readily transformed to methyl groups with broad functional group and heterocycle tolerance.

Quaternary Trimethyl Chitosan Chloride Capped Bismuth Nanoparticles with Positive Surface Charges: Catalytic and Antibacterial Activities

Quaternary trimethyl chitosan-stabilized bismuth nanoparticles (QTMC-BiNPs) with positive surface charges were uniquely synthesized and fully characterized. In the synthesis, Quaternary Trimethyl Chitosan (QTMC), a water-soluble derivative of chitosan (CTS) was prepared using two-step reductive methylation. The new biopolymeric functionalized ligand was further used as capping agent for the synthesis of QTMC-BiNPs which was applied as antibacterial and catalytic agents. The reaction was carried out at room temperature without the use of energy consuming or high-cost instruments. The QTMC and nanocomposites were characterized by proton nuclear magnetic resonance (1H NMR), attenuated total reflection Fourier-transform infrared, UV–visible, X-ray diffraction, X-ray photoelectron spectroscopy and energy dispersive X-ray spectroscopic techniques. The topology and morphology of the composites were examined with scanning electron microscopy and high-resolution transmission electron microscopy. Thermogravimetric and differential thermal gravimetric analysis were also conducted. The degree of quaternization and degree of dimethylation values of 63.33 and 11.75%, respectively obtained for QTMC confirmed that the main product is a quaternary derivative. The average particle size of QTMC-BiNPs was evaluated to be between 30 and 45 nm. The QTMC-BiNPs revealed clear and uniform lattice fringes with an estimated interplanar d-spacing of 0.32 nm confirming the formation of highly crystalline nanocomposites. A further insight into the antibacterial activities of this nanomaterial were carefully examined on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) using resazurin based microdilution method for Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). The obtained results revealed that both bacteria pathogens were effectively inhibited/killed by the QTMC-BiNPs at very low concentrations. The MIC of 15.63 and 125 µg/mL were recorded against E. coli and S. aureus, respectively while the MBC of 31.25 and 500.00 µg/mL were estimated against E. coli and S. aureus, respectively. An extensive evaluation of the catalytic capability of the nanocomposites towards the reduction of 4-nitrophenol to 4-aminophenol was also carried out with highly promising result.

Reductive Methylation of Homogeneous Primary β-Lauryl/myristyl 7/3 Polyethyleneoxy n = 3-18 Ethylamines under Phase-Transfer Catalysis Conditions.

Homogeneous tertiary N,N-dimethyl-N-β-lauryl/myristyl 7/3 polyethyleneoxy n = 3–18 ethylamines, LM(EO)nAT, are niche intermediates in the synthesis of homogeneous N-alkyl (C1–C18)-N,N-dimethyl-N-β-lauryl/myristyl 7/3 polyethyleneoxy n = 3–18 ethylammonium chlorides (unitary degree of oligomerization of ethylene oxide in the polyoxyethylene chain). This paper synthetically presents the dependence of the reductive methylation yields of homogeneous primary β-lauryl/myristyl 7/3 polyethyleneoxy n = 3–18 ethylamines, LM(EO)nAP, on the reaction time (10–90 min), the temperature (70 °C), the molar ratio formic aldehyde /LM(EO)nAP (1.1/1–2.5/1), the molar ratio HCOOH/LM(EO)nAP (5/1), the degree of oligomerization of ethylene oxide in the homogeneous polyoxyethylene chain in the 3,6,9,12,18 series, and the structure of the phase-transfer catalysts. The steric effects of hydrophobic groups CH3 and C18H37 grafted onto the ammonium function, and the micellar phenomena in the vicinity of their critical micellar concentration, directly proportional to the homogeneous degree of oligomerization, were highlighted. In all cases, a steady increase in reductive methylation yields was observed, with even quantitative values obtained. The high purity of the homologous series LM(EO)nAT will allow their personalization as reference structures for the study of the evolution of basic colloidal characteristics useful in forecasting technological applications. LM(EO)nAP were obtained either by direct amidoethylation (nucleophilic addition under basic catalysis of homogeneous lauryl/myristyl 7/3 polyethoxylated n = 3, 6, 9, 12, 18 alcohols, LM(EO)nOH, to acrylamide monomer) or by cyanoethylation of LM(EO)nOH under basic catalysis at 25–50 °C, in the presence of Fe2+ cations as oligomerization/polymerization inhibitor, followed by partial acid hydrolysis of homogeneous β-alkyl (C12H25/C14H29) 7/3 polyethyleneoxy n = 3, 6, 9, 12, 18 propionitriles, LM(EO)nPN, to β-alkyl (C12H25/C14H29) 7/3 polyethyleneoxy n = 3, 6, 9, 12, 18 propionamides, LM(EO)nPD, which led to LM(EO)nAP by Hoffmann degradation. Homogeneous higher tertiary polyetheramines LM(EO)nAT were structurally characterized.