Nuclear Magnetic Resonance Analysis Research Articles

Structural and dynamic insights into the activation of the μ-opioid receptor by an allosteric modulator

G-protein-coupled receptors (GPCRs) play pivotal roles in various physiological processes. These receptors are activated to different extents by diverse orthosteric ligands and allosteric modulators. However, the mechanisms underlying these variations in signaling activity by allosteric modulators remain largely elusive. Here, we determine the three-dimensional structure of the μ-opioid receptor (MOR), a class A GPCR, in complex with the Gi protein and an allosteric modulator, BMS-986122, using cryogenic electron microscopy. Our results reveal that BMS-986122 binding induces changes in the map densities corresponding to R1673.50 and Y2545.58, key residues in the structural motifs conserved among class A GPCRs. Nuclear magnetic resonance analyses of MOR in the absence of the Gi protein reveal that BMS-986122 binding enhances the formation of the interaction between R1673.50 and Y2545.58, thus stabilizing the fully-activated conformation, where the intracellular half of TM6 is outward-shifted to allow for interaction with the Gi protein. These findings illuminate that allosteric modulators like BMS-986122 can potentiate receptor activation through alterations in the conformational dynamics in the core region of GPCRs. Together, our results demonstrate the regulatory mechanisms of GPCRs, providing insights into the rational development of therapeutics targeting GPCRs.

Relaxation Editing in NMR Using a New Two-Dimensional Long-Lived Coherence Method for Mixture Analysis.

The exploration of metabolomics and targeted segments of proteins stands as a pivotal facet of Nuclear Magnetic Resonance (NMR) analysis, furnishing valuable insights into molecular architectures and potential therapeutic applications. The issue of spectral congestion frequently presents challenges in ascribing distinct peaks within the confines of both one-dimensional (1D) and two-dimensional (2D) NMR spectra. Numerous strategies have been proposed to resolve specific resonances in NMR spectra differentially. Among these approaches, relaxation editing emerges as a viable solution. In the realm of relaxation phenomena within NMR, Long-Lived States (LLS) and Long-Lived Coherences (LLC) manifest as promising phenomena, offering enhanced relaxation lifetimes in comparison to the traditional longitudinal (T1) and transverse (T2) relaxation times for coupled nuclear spins. Notably, LLC presents a pathway to attenuate uncoupled high-intensity peaks, effectively diminishing their impact. The foundation of this technique rests upon the premise that the relaxation lifetime in the rotating frame (T1ρ) remains smaller than TLLC. In pursuit of refining spectral assignments within complex mixtures, we introduce a new pulse sequence tailored for LLC Total Correlation Spectroscopy (LLC-TOCSY). This demonstrates efficacy in extracting LLC signals within configurations involving multiple coupled spins, thereby decluttering the spectrum and enhancing the accuracy of peak assignments. To validate the effectiveness of this method, a collection of samples was subjected to scrutiny, yielding compelling results.

Anaerobic sludge from a full-scale biodigester: physico-chemical and organic characterization, and principal component analysis (PCA)

The biological transformation of the organic fraction of municipal solid waste into bioenergy yields a solid digested material known as anaerobic sludge, comprising a mixture of organic and inorganic substances. This sludge holds significant potential as a renewable resource for agricultural reuse. However, there is also the potential risk of contamination from this material in receiving environments. Therefore, it is crucial to assess the characteristics of this material to ensure compliance with local regulations regarding its soil application, aiming to avoid adverse environmental impacts and ensure proper and viable disposal. In this study, conducted to explore the characteristics of bottom sludge collected from a pilot-scale anaerobic biodigester, various parameters were analyzed. The biodigester in question had a capacity of 1300 liters and a useful volume of 1000 liters, digesting food waste and water hyacinth (Eichhornia crassipes) in a ratio of 20% in terms of volatile solids from the macrophyte to the volatile solids of the food waste. A multivariate analysis was conducted on the following parameters: metals (Al, Ba, Cd, Cr, Cu, Fe, Mn, Mo, Pb, Se, V, Zn, Na, and K); total, fixed, and volatile solids; alkalinity; volatile fatty acids; ammonia; Kjeldahl nitrogen (NTK); and organic, inorganic, and total carbon (COT), to observe possible correlations among these parameters. Additionally, the sludge underwent nuclear magnetic resonance (NMR) analysis for detection and characterization of organic compounds. This approach provided a deeper understanding of the properties of anaerobic sludge produced strictly under the aforementioned conditions, contributing to further research in the field.

Lignin Degradation by Klebsiella aerogenes TL3 under Anaerobic Conditions

Lignin, the largest non-carbohydrate component of lignocellulosic biomass, is also a recalcitrant component of the plant cell wall. While the aerobic degradation mechanism of lignin has been well-documented, the anaerobic degradation mechanism is still largely elusive. In this work, a versatile facultative anaerobic lignin-degrading bacterium, Klebsiella aerogenes TL3, was isolated from a termite gut, and was found to metabolize a variety of carbon sources and produce a single kind or multiple kinds of acids. The percent degradation of alkali lignin reached 14.8% under anaerobic conditions, and could reach 17.4% in the presence of glucose within 72 h. Based on the results of infrared spectroscopy and 2D nuclear magnetic resonance analysis, it can be inferred that the anaerobic degradation of lignin may undergo the cleavage of the C-O bond (β-O-4), as well as the C-C bond (β-5 and β-β), and involve the oxidation of the side chain, demethylation, and the destruction of the aromatic ring skeleton. Although the anaerobic degradation of lignin by TL3 was slightly weaker than that under aerobic conditions, it could be further enhanced by adding glucose as an electron donor. These results may shed new light on the mechanisms of anaerobic lignin degradation.

From Bulk to Interface: Solvent Exchange Dynamics and Their Role in Ion Transport and the Interfacial Model of Rechargeable Magnesium Batteries.

Multivalent battery chemistries have been explored in response to the increasing demand for high-energy rechargeable batteries utilizing sustainable resources. Solvation structures of working cations have been recognized as a key component in the design of electrolytes; however, most structure-property correlations of metal ions in organic electrolytes usually build upon favorable static solvation structures, often overlooking solvent exchange dynamics. We here report the ion solvation structures and solvent exchange rates of magnesium electrolytes in various solvents by using multimodal nuclear magnetic resonance (NMR) analysis and molecular dynamics/density functional theory (MD/DFT) calculations. These magnesium solvation structures and solvent exchange dynamics are correlated to the combined effects of several physicochemical properties of the solvents. Moreover, Mg2+ transport and interfacial charge transfer efficiency are found to be closely correlated to the solvent exchange rate in the binary electrolytes where the solvent exchange is tunable by the fraction of diluent solvents. Our primary findings are (1) most battery-related solvents undergo ultraslow solvent exchange coordinating to Mg2+ (with time scales ranging from 0.5 μs to 5 ms), (2) the cation transport mechanism is a mixture of vehicular and structural diffusion even at the ultraslow exchange limit (with faster solvent exchange leading to faster cation transport), and (3) an interfacial model wherein organic-rich regions facilitate desolvation and inorganic regions promote Mg2+ transport is consistent with our NMR, electrochemistry, and cryogenic X-ray photoelectron spectroscopy (cryo-XPS) results. This observed ultraslow solvent exchange and its importance for ion transport and interfacial properties necessitate the judicious selection of solvents and informed design of electrolyte blends for multivalent electrolytes.

Synthesis, characterization, cytotoxicity study, interaction with DNA and topoisomerase IIα of square-planar complexes with thiosemicarbazones

In this study, we report the synthesis and characterization of eight new palladium complexes and a nickel complex containing thiosemicarbazone ligands derived from chalcones and benzalketones. The techniques of Infrared and UV–visible spectroscopy, Nuclear Magnetic Resonance, Mass Spectrometry, and Elemental Analysis fully characterized the compounds. Crystallography was successful for five complexes and two ligands, allowing for X-ray Diffraction analysis. Molecular docking studies and assays involving DNA interaction via agarose gel electrophoresis and UV–Vis titration revealed no DNA interaction or inhibition of topoisomerase IIα enzyme activity. However, these compounds demonstrated potent cytotoxic effects (IC50) against breast tumor (MCF-7) and prostate (DU-145) cell lines, as well as non-tumor prostate (PNT2) cell lines. These findings highlight the relevance of structural variations and the metallic center's nature in achieving cytotoxic effects, offering insights into the development of bioactive compounds.

Antibacterial activity of Dioscorea bulbifera Linn. extract and its active component flavanthrinin against skin-associated bacteria

BackgroundDioscorea bulbifera Linn. has been used for wound care in Thailand. However, a comprehensive evaluation of its antibacterial activity is required. This study aimed to investigate the antibacterial efficacy of D. bulbifera extract against skin-associated bacteria and isolate and characterize its active antibacterial agent, flavanthrinin.MethodsAir-dried bulbils of D. bulbifera were pulverised and extracted with hexane, dichloromethane, ethyl acetate, methanol, ethanol, and distilled water; vacuum filtered; concentrated; freeze-dried; and stored at -20 ºC. Antibacterial activity of the extracts was assessed using microdilution techniques against several skin-associated bacteria. Thin-layer chromatography (TLC) bioautography was used to identify the active compounds in the extract, which were fractionated by column chromatography and purified by preparative TLC. The chemical structures of the purified compounds were analysed using nuclear magnetic resonance (NMR). The cytotoxicity of the extract and its active compounds was evaluated in Vero cells.ResultsThe ethyl acetate extract exhibited distinct inhibition zones against bacteria compared to other extracts. Therefore, the ethyl acetate extract of D. bulbifera in the ethyl acetate layer was used for subsequent analyses. D. bulbifera extract exhibited antibacterial activity, with minimum inhibitory concentrations (MICs) of 0.78–1.56 mg/mL. An active compound, identified through TLC-bioautography, demonstrated enhanced antibacterial activity, with MICs of 0.02–0.78 mg/mL. NMR analysis identified this bioactive compound as flavanthrinin. Both D. bulbifera extract and flavanthrinin-containing fraction demonstrated potent antibacterial activity against Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and S. epidermidis. The flavanthrinin containing fraction demonstrated low cytotoxicity against Vero cells, showing CC50 values of 0.41 ± 0.03 mg/mL. These values are lower than the MIC value, indicating that this fraction is safer than the initial ethyl acetate extract.ConclusionsDioscorea bulbifera extract and its bioactive component flavanthrinin demonstrated significant antibacterial activity against the skin-associated bacteria Staphylococci, including MRSA. Flavanthrinin has potential as a complementary therapeutic agent for managing skin infections owing to its potent antibacterial effects and low cytotoxicity.

Evaluation of Modified Asphalt Binders with Soybean Oil-Based Polymers: Preparation Temperature and Rheological Characterization

To investigate the feasibility of bio-based polymers in the modification of base asphalt binder, two soybean oil-based polymers (PAESO) were synthesized from bio-monomers with different functionalities via free radical polymerization. The characterization of both PAESOs was conducted using Hydrogen nuclear magnetic resonance and Gel Permeation Chromatography analyses, the results showed that the bio-monomer with a higher functionality exhibited a stronger tendency towards self-polymerization, resulting in the polymer with a high conversion rate (44.44 %) and a slightly lower molecular weight (14.76 kDa). The optimal mixing temperature for preparing PAESO-modified asphalt binders was determined through Molecular Dynamics (MD) simulation and Laser Scanning Confocal Microscopy. The MD simulation results indicated that PAESO-modified asphalt models had reached the most stable configurations at a mixing temperature of 160 °C, as evidenced by fewer and more homogeneous distributed small bright spots observed in fluorescence images. This suggested that the outstanding compatibility between PAESO and asphalt binders could be obtained at the optimal mixing temperature of 160 °C. Furthermore, the rheological characteristics of asphalt binders modified with PAESO were examined through Dynamic Shear Rheometer and Bending Beam Rheometer tests. The results revealed that PAESO mainly exhibited a softening effect by significantly enhancing the flexibility, stress relaxation, fatigue, and thermal cracking resistance of the base asphalt binders at intermediate and low temperatures. PAESO-L modified asphalt manifested elastic properties akin to those observed in polymer-modified asphalt within the low-frequency and high-temperature domain. Additionally, PAESO derived from monomers with a higher functionality presented notable benefits in enhancing thermal and fatigue cracking resistance than that of PAESO with a lower functionality. These findings provide valuable insights for the preparation and development of bio-polymer additives for use in asphalt materials.

High throughput identification of carbonyl compounds in natural organic matter by directional derivatization combined with ultra-high resolution mass spectrometry

Carbonyl compounds are important components of natural organic matter (NOM) with high reactivity, so that play a pivotal role in the dynamic transformation of NOM. However, due to the lack of effective analytical methods, our understanding on the molecular composition of these carbonyl compounds is still limited. Here, we developed a high-throughput screening method to detect carbonyl molecules in complex NOM samples by combining chemical derivatization with electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR-MS). In six different types of dissolved organic matter (DOM) samples tested in this study, 20%-30% of detected molecules contained at least one carbonyl group, with relative abundance accounted for 45% to 70%. These carbonyl molecules displayed lower unsaturation level, lower molecular weight, and higher oxidation degree compared to non-carbonyl molecules. More importantly, the measured abundances of carbonyl molecules were consistent with the results of 13C nuclear magnetic resonance (NMR) analysis. Based on this method, we found that carbonyl molecules can be produced at DOM-ferrihydrite interface, thus playing a role in shaping the molecular diversity of DOM. This method has broad application prospects in screening carbonyl compounds from complex mixtures, and the same strategy can be used to directional identification of molecules with other functional groups as well.

Quantitative Characterization of Crude Oil Mobilization in Microscopic Pores During Forward and Reverse Flooding in Tight Sandstone Reservoirs Using Various Fracturing Fluid Systems

In the transition from conventional to unconventional oil and gas exploration, the focus on tight sandstone reservoirs has grown due to their complex microscopic pore structures, narrow pore throats, and high capillary pressures. These characteristics often lead to the retention of fracturing fluids post-fracturing, hindering fluid recovery and reducing crude oil extraction efficiency. To address these issues, this study employed a combination of microscopic displacement experiments and nuclear magnetic resonance (NMR) analyses to investigate oil displacement mechanisms at the microscale in tight sandstone reservoirs. The experimental setup involved testing three core samples using distinct fracturing fluid systems: a guar gum-based system (System I), an EM30+ + guar gum water-based system (System II), and a CNI (Carbon Nanotechnologies Inc) nano variable-viscosity slick water-based system (System III). The study aimed to assess the impact of these different fracturing fluid systems on oil displacement efficiency in both forward and reverse directions. Results revealed that forward oil displacement efficiencies for Systems I to III were 26.86%, 33.3%, and 33.43%, respectively. In contrast, reverse displacement efficiencies were slightly higher at 28.9%, 37.5%, and 38.34%. Overall, Systems II and III outperformed System I in terms of oil displacement efficiency. The study also found that in cores with large pores, forward displacement predominantly mobilized crude oil, while reverse displacement was more effective in smaller pores. This trend was consistent in cores exhibiting bimodal pore distributions. However, in unimodal cores, small pores were significantly impacted in both directions. Notably, System III demonstrated a remarkable efficiency in mobilizing crude oil within smaller pores.

Variation pattern in the macromolecular (cellulose, hemicelluloses, lignin) composition of cell walls in Pinus tabulaeformis tree trunks at different ages as revealed using multiple techniques

The plant cell wall is a complex, heterogeneous structure primarily composed of cellulose, hemicelluloses, and lignin. Exploring the variations in these three macromolecules over time is crucial for understanding wood formation to enhance chemical processing and utilization. Here, we comprehensively analyzed the chemical composition of cell walls in the trunks of Pinus tabulaeformis using multiple techniques. In situ analysis showed that macromolecules accumulated gradually in the cell wall as the plant aged, and the distribution pattern of lignin was opposite that of polysaccharides, and both showed heterogenous distribution patterns. In addition, gel permeation chromatography (GPC) results revealed that the molecular weights of hemicelluloses decreased while that of lignin increased with age. Two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance (2D-HSQC NMR) analysis indicated that hemicelluloses mainly comprised galactoglucomannan and arabinoglucuronoxylan, and the lignin types were mainly comprised guaiacyl (G) and p-hydroxyphenyl (H) units with three main linkage types: β-O-4, β-β, and β-5. Furthermore, the C-O bond (β-O-4) signals of lignin decreased while the C-C bonds (β-β and β-5) signals increased over time. Taken together, these findings shed light on wood formation in P. tabulaeformis and lay the foundation for enhancing the processing and use of wood and timber products.

α-Zr(HPO4)2·H2O: As an effective and recoverable catalyst for the synthesis of a family of quinoxalines in semi-aqueous conditions

In our work, we have established a straightforward and friendly process for the preparation of a family of quinoxaline. This method relies on using α-Zr(HPO4)2·H2O as a catalyst and an ethanol/water mixture as an ecological solvent. Firstly, we prepared α-Zr(HPO4)2·H2O by a simple hydrothermal procedure and conducted comprehensive characterization employing various analytical methods. On the other hand, we produced the quinoxalines by means of a condensation reaction between 1,2-dicarbonyl compounds and various 1,2-diamines. The synthesized quinoxaline and its derivative products were characterized by melting point, proton, and carbon nuclear magnetic resonance analysis. As a result, the optimal reaction conditions that resulted in a high yield (97–98 %) were an amount of catalyst of 0.03 g, a stirring time of 8 min, a volume of solvent of 2 mL, and a reaction temperature of 50 °C. This approach is advantageous due to its simple and eco-friendly procedure, green solvent, quick reaction time, low cost, and high yields. Additionally, it offers the possibility to recycle the catalyst for a minimum of five cycles with no decrease in its catalytic activity.

Synthesis, characterization, and antibacterial studies of some of first transition series metals and zinc complexes with mixed ligands of trimethoprim-isatin and nitrogen base

Complexes of mixed-ligand Schiff bases [trimetho-prim (TMP) with isatin] (L1) and nitrogen bases [2,2′-bipyridine (bipy)] (L2), [1,10-phenanthroline (phen) (L3)] with metal chlorides: M = Cr3+, Mn2+, Co2+, Ni2+, Cu2+, Zn2+ were prepared and characterized by metal content determination (%), FT-IR, electronic, nuclear magnetic resonance, mass spectroscopy, magnetic susceptibility, conductance measurements and elemental analysis (CHN). The bacterial activity of the ligands and their metal compounds was evaluated via various microorganisms. Infrared spectra of Schiff base showed an azomethine peak shifted in complexes that coincide synthesis and coordination of Schiff base through the nitrogen atom of azomethine C=N) and the oxygen atom of the carbonyl group of L1 with metal ions. L2, L3 (bipy, phen) coordinated through the di imine nitrogen atoms. Conductance dimensions suggested the non-conductance nature of all the compounds except Cr(III) complexes, this outcome fits in with the molar conductance value estimated for 1:1 electrolyte in DMF. Magnetic susceptibility analysis and electronic spectra indicated an octahedral geometry for all complexes. Biological study for prepared compounds have been evaluated towards pathogenic microbes Staphylococcus aureus and Bacillus subtilis. Some compounds have stronger antibacterial action than Schiff base towards tested microbes. KEY WORDS: Antimicrobial, Heterocyclic compounds, Schiff base, Trimethoprim drug, 2,2-Bipyridyl, 1,10-Phenanthroline Bull. Chem. Soc. Ethiop. 2024, 38(4), 1013-1025.                                                            DOI: https://dx.doi.org/10.4314/bcse.v38i4.16

A novel integrated membrane bioreactor-stirred tank bioreactor system for simultaneous biodegradation of bisphenol A contaminated wastewater and biopolyol production: An approach towards circular bioeconomy

The present work examined the biodegradation of bisphenol A (BPA) under batch and continuous modes using hydrocarbonoclastic bacterium, Pseudomonas aeruginosa PR3. The strain showed great biodegradation potential under batch shake flask and stirred tank reactor. Bacterial growth was considerably inhibited at initial BPA concentrations > 50 mg/L. Further, membrane bioreactor (MBR) for BPA biodegradation was also evaluated under different inlet BPA concentration and hydraulic retention time (HRT) and the maximum BPA removal efficiency of 94.4 % was attained with an influent BPA concentration of 25 mg/L at 5 d of HRT. MBR also showed better performance under shock loading conditions. Identification of BPA biodegraded pathways was also performed using liquid chromatography-mass spectrometry, elucidating the different metabolites and end products formed. A subsequent enhancement in BPA removal efficieny was also observed owing to the microfiltration system of MBR that was applied to separate bacterial biomass from effluent. The maximum removal efficiency was further enhanced to 98.5 % under the same operational schedule. As the strain has the ability to produce extracellular diol synthase enzyme which transforms oleic acid into the biopolyol 7,10-dihydroxy-8(E)-octadecenoic acid, the effluent containing the enzymes was supplemented with 1 % (v/v) oleic acid under whole cell and cell-free approaches. Cell-free technique showed an enhanced bioconversion efficiency in comparison to the traditional whole cell approach. Further, the produced biopolyol was characterized using Fourier transform infrared spectroscopy, thermogravimetry, differential scanning calorimetry, and 1H nuclear magnetic resonance analyses and also depicted the antibacterial activity.

Carboxymethyl cellulose-based rotigotine nanocrystals-loaded hydrogel for increased transdermal delivery with alleviated skin irritation

Transdermal rotigotine (RTG) therapy is prescribed to manage Parkinson's disease (Neupro® patch). However, its use is suffered from application site reactions. Herein, drug nanocrystalline suspension (NS)-loaded hydrogel (NS-HG) employing polysaccharides simultaneously as suspending agent and hydrogel matrix was constructed for transdermal delivery, with alleviated skin irritation. RTG-loaded NS-HG was prepared using a bead-milling technique, employing sodium carboxylmethyl cellulose (Na.CMC) as nano-suspending agent (molecular weight 90,000 g/mol) and hydrogel matrix (700,000 g/mol), respectively. NS-HG was embodied as follows: drug loading: ≤100 mg/mL; shape: rectangular crystalline; crystal size: <286.7 nm; zeta potential: −61 mV; viscosity: <2.16 Pa·s; and dissolution rate: >90 % within 15 min. Nuclear magnetic resonance analysis revealed that the anionic polymers bind to RTG nanocrystals via charge interaction, affording uniform dispersion in the matrix. Rodent transdermal absorption of RTG from NS-HG was comparable to that from microemulsions, and proportional to drug loading. Moreover, NS-HG was skin-friendly; erythema and epidermal swelling were absent after repeated application. Further, NS-HG was chemically stable; >95 % of the drug was preserved up to 4 weeks under long term (25 °C/RH60%), accelerated (40 °C/RH75%), and stress (50 °C) storage conditions. Therefore, this novel cellulose derivative-based nanoformulation presents a promising approach for effective transdermal RTG delivery with improved tolerability.

Brønsted Acid-Promoted Cyclodimerization of α,β-Unsaturated γ-Ketoesters: Construction of Fused Pyrano-ketal-lactones and γ-Ylidene-butenolides.

Unprecedented MsOH-promoted diastereoselective cascade dimerization and intramolecular lactonization of readily accessible α,β-unsaturated γ-ketoesters are presented. The results obtained in this work, control experiments, and density functional theory (DFT) calculations suggested that the initial enolization and E to Z isomerization/equilibration of olefin (C=C) of substrate α,β-unsaturated γ-ketoesters give a Z-isomer preferentially over an E-isomer. Subsequently, the Z-isomer undergoes intermolecular annulation with α,β-unsaturated γ-ketoesters via domino Michael addition/ketalization/lactonization steps to furnish fused tetracyclic pyrano-ketal-lactone. However, the Z-isomer prefers intramolecular trans-esterification in a competing pathway and gives bicyclic γ-ylidene-butenolide. The key features of this work include simple Brønsted acid catalysis, the formation of three bonds, two rings, and three contiguous stereogenic centers in a single step, DFT calculations, and the assignment of relative stereochemistry through X-ray diffraction (XRD) and two-dimensional (2D) nuclear magnetic resonance (NMR) analyses.

Efficient enzymatic synthesis of vitamin E succinate using an organic solvent‐stable immobilized lipase

AbstractVitamin E succinate has gained substantial attention as a potential therapeutic agent for cancer treatment due to its biomedical activities. One of the prominent methods of synthesizing vitamin E succinate is through enzymatic processes, which, although advantageous, presents inherent challenges related to optimization, scalability, and particularly, the poor stability of lipases in organic solvents. Our study addresses these challenges by conducting a comprehensive comparative analysis between Lipase UM1 and three other immobilized commercial lipases, demonstrating Lipase UM1's enhanced resistance to organic solvents and its superior efficiency in vitamin E succinate production. Further optimization experiments with Lipase UM1 led to an unprecedented conversion of 99%. Additionally, we scaled the reaction to a proof‐of‐concept industrial level. The synthesized product was verified using Fourier transform infrared spectroscopy and nuclear magnetic resonance analysis, ensuring its quality and consistency. This study validates Lipase UM1 as an efficient catalyst for vitamin E succinate synthesis, offering a promising avenue for industrial production with potential applications in cancer therapy and beyond.

Preparation of low estimated glycemic index (eGI) chestnut flours using enzymatic debranching and heat moisture treatment

A low estimated glycemic index (eGI) chestnut flour was prepared using a combination of pullulanase debranching and heat moisture treatment (HMT). The modified samples had a lower eGI range of 49.1–53.0 compared to native chestnut flour (NCF) which had an eGI of 67.3. Furthermore, the dual treatment was found to be more effective than a single treatment. The total starch content decreased from 57.0 to 47.1 g/100 g after debranching and HMT, resulting in an increase in amylose content from 30.6 % to 48.5 %. A 1H NMR (nuclear magnetic resonance) analysis revealed that modified flours showed a reduction in α-1,6-glycosidic bonds. NCF exhibited a C-type crystal structure, while the debranched samples showed B+V-type crystallinity with an increased peak at 20°(2θ). Debranching increased the proportion of short linear chains, which facilitated the formation of amylose-lipid complexes. HMT resulted in partial starch gelatinization and the breakage of amylopectin. These findings emphasize the potential applications of debranching and HMT in producing chestnut flour with a low eGI, which can further be used in functional food products.

Structural characterization and anti-inflammatory activities of a purified polysaccharide from fruits remnants of Alpinia zerumbet (Pers.) Burtt. et Smith

We reported here an interesting source of Alpinia zerumbet Polysaccharides (named AZPs) from the residues after extracting essential oil by steam distillation from Alpinia zerumbet fructus. After a series of purifications, a homogeneous polysaccharide (AZP-2) of molecular weight 1.25 × 105 Da was obtained. Structure, anti-inflammatory activity, and anti-inflammatory mechanism were investigated. AZP-2 was mainly composed of galactose, arabinose, xylopyranose, glucose, and galacturonic acid. The main linkage structure of AZP-2 was determined after integrating the nuclear magnetic resonance (NMR) and methylation analysis, and the structure was comparatively complex. The results indicated that AZP-2 significantly decreased the production of NO and ROS in the inflammatory model established by lipopolysaccharide (LPS) stimulated RAW264.7, particularly at the concentration of 200 μg/mL. Furthermore, AZP-2 significantly modulated the secretion of both pro-inflammatory and anti-inflammatory cytokines. Notably, the mechanism of AZP-2 exhibiting inhibitory effects was related to regulating the NF-κB signaling pathway. Overall, AZP-2 could be used as a potential anti-inflammatory agent for further in-depth studies.

The Effects of Four Different Thawing Methods on Quality Indicators of Amphioctopus neglectus.

Amphioctopus neglectus is a species of octopus that is favored by consumers due to its rich nutrient profile. To investigate the influence of different thawing methods on the quality of octopus meat, we employed four distinct thawing methods: air thawing (AT), hydrostatic thawing (HT), flowing water thawing (FWT), and microwave thawing (MT). We then explored the differences in texture, color, water retention, pH, total volatile basic nitrogen (TVB-N), total sulfhydryl content, Ca2+-ATPase activity, and myofibrillar protein, among other quality indicators in response to these methods, and used a low-field nuclear magnetic resonance analyzer to assess the water migration that occurred during the thawing process. The results revealed that AT had the longest thawing time, leading to oxidation-induced protein denaturation, myofibrillar protein damage, and a significant decrease in water retention. Additionally, when this method was utilized, the content of TVB-N was significantly higher than in the other three groups. HT, to a certain extent, isolated the oxygen in the meat and thus alleviated protein oxidation, allowing higher levels of Ca2+-ATPase activity, sulfhydryl content, and springiness to be maintained. However, HT had a longer duration: 2.95 times that of FWT, resulting in a 9.84% higher cooking loss and a 28.21% higher TVB-N content compared to FWT. MT had the shortest thawing time, yielding the lowest content of TVB-N. However, uneven heating and in some cases overcooking occurred, severely damaging the protein structure, with a concurrent increase in thawing loss, W value, hardness, and shear force. Meanwhile, FWT improved the L*, W* and b* values of octopus meat, enhancing its color and water retention. The myofibrillar protein (MP) concentration was also the highest after FWT, with clearer subunit bands in SDS-PAGE electrophoresis, indicating that less degradation occurred and allowing greater springiness, increased Ca2+-ATPase activity, and a higher sulfhydryl content to be maintained. This suggests that FWT has an inhibitory effect on oxidation, alleviating protein oxidation degradation and preserving the quality of the meat. In conclusion, FWT outperformed the other three thawing methods, effectively minimizing adverse changes during thawing and successfully maintaining the quality of octopus meat.