HSQC Research Articles

In vitro anticancer and antibacterial potentials of selected medicinal plants and isolation and characterization of a natural compound from Withania coagulans.

In the current study, five different plants, Syzygium Cumini, Fagonia cretica, Acacia modesta, Withania coagulans, and Olea europaea aqueous extracts were prepared and applied against the anticancer and antibacterial activities. It was observed that O.Europaea extract shows the highest anticancer activity with cell viability of 21.5%. All the five plants extract was also used against the inhibition of Bacillus subtilis where O.Europaea extract shows a promising inhibitory activity of 3.2cm followed by W.coagulans. Furthermore, W.coagulans was subjected to the process of column chromatography as a result a withanolide was isolated. The fast atom bombardment mass spectrometry (FAB-MS) and high resolution fast atom bombardment (HRFAB-MS) [M+1] indicated molecular weight at m/z 453 and molecular formula C28H37O5. The UV-Vis. spectrum shows absorbance at 210nm suggesting the presence of conjugated system, and Fourier-transform infrared spectroscopy (FTIR) was recorded to explore the functional groups. Similarly, 1D and 2D NMR spectroscopy techniques such as 1H, 13C NMR, correlation spectroscopy (COSY-45°), heteronuclear single quantum correlation (HSQC), heteronuclear multiple bond correlation (HMBC) and Nuclear Overhauser effect Spectroscopy (NOESY) techniques was carried out to determine the unknown natural product. The collective data of all these techniques established the structure of the unknown compound and recognized as a withanolide.

Unusual ent-Kaurane Diterpenes from the Coffea Cultivar S288 Coffee Beans and Molecular Docking to α-Glucosidase.

A total of 11 new (1-11) and 2 known (12 and 13) ent-kaurane diterpene derivatives were identified from the roasted beans of Coffea cultivar S288. Their structures were established by extensive spectroscopic analysis, including one- and two-dimensional nuclear magnetic resonance (heteronuclear single-quantum correlation, heteronuclear multiple-bond correlation, correlation spectroscopy, and rotating-frame Overhauser enhancement spectroscopy), high-resolution electrospray ionization mass spectrometry, and X-ray analyses. Cafespirone acid A (1) represents the first example of diterpene featuring a spirocyclic skeleton constructed from a 6/6/5 tricyclic system. Cafeane acid A (2) possesses a 6/6/6/5 tetracyclic system as a result of the C/D ring rearrangement. Furthermore, compounds 1-12 were evaluated for their α-glucosidase inhibitory activity. The results showed that compounds 2, 4, 5, 6, 7, 10, and 11 had a moderate inhibitory effect on α-glucosidase, and half-maximal inhibitory concentration values of compounds 4, 6, 7, and 10 were 18.76 ± 1.46, 4.88 ± 0.03, 12.35 ± 0.91, and 12.64 ± 0.59 μM, respectively, compared to the positive control acarbose (60.71 ± 16.45 μM). Additionally, the molecular docking experiments showed that the carbonyl group at C-19 of compounds 4, 6, and 7 formed strong hydrogen bonds with ARG315, which may make them have moderate inhibitory activity.

Theoretical and experimental investigation of four ionic liquids for their ability to prevent carbon steel corrosion in 1 M HCl

The inhibitory mechanism of four ionic liquids (ILs) generated from imidazolium is reported in this study. The goal of the research was to see how the cation structure of ILs affected the corrosion inhibition of carbon steel in 1 M HCl. The chemical structure of ILs was verified by elements analysis, Fourier-transform-infrared (FTIR), thermo gravimetric (TGA), nuclearmagnetic resonance (1HNMR, D2O, 13C NMR) and hetero-nuclear single-quantum correlation (HSQC). The inhibitory efficiency was measured using mass loss and electrochemical tests and varied from 62.5% to 97.2%. The Langmuir model accurately defined their adsorption on a steel substrate. Adsorption of novel ILs on steel substrates is a mixed physisorption and chemisorption method. To ensure the mechanism of adsorption of current ionic liquids, FTIR and ultraviolet (UV) spectra were employed.

Structural stability of Cutibacterium acnes acyl carrier protein studied using CD and NMR spectroscopy

To survive in diverse environments, bacteria adapt by changing the composition of their cell membrane fatty acids. Compared with aerobic bacteria, Cutibacterium acnes has much greater contents of branched-chain fatty acids (BCFAs) in the cell membrane, which helps it survive in anaerobic environments. To synthesize BCFAs, C. acnes acyl carrier protein (CaACP) has to transfer growing branched acyl intermediates from its hydrophobic cavity to fatty acid synthases. CaACP contains an unconserved, distinctive Cys50 in its hydrophobic pocket, which corresponds to Leu in other bacterial acyl carrier proteins (ACPs). Herein, we investigated the substrate specificity of CaACP and the importance of Cys50 in its structural stability. We mutated Cys50 to Leu (C50L mutant) and measured the melting temperatures (Tms) of both CaACP and the C50L mutant by performing circular dichroism experiments. The Tm of CaACP was very low (49.6 °C), whereas that of C50L mutant was 55.5 °C. Hydrogen/deuterium exchange experiments revealed that wild-type CaACP showed extremely fast exchange rates within 50 min, whereas amide peaks of the C50L mutant in the heteronuclear single quantum coherence spectrum remained up to 200 min, thereby implying that Cys50 is the key residue contributing to the structural stability of CaACP. We also monitored chemical shift perturbations upon apo to holo, apo to butyryl, and apo to isobutyryl conversion, confirming that CaACP can accommodate isobutyryl BCFAs. These results provide a preliminary understanding into the substrate specificity of CaACPs for the production of BCFAs necessary to maintain cell membrane fluidity under anaerobic environments.

Alkylation modification for lignin color reduction and molecular weight adjustment

The application of industrial kraft lignin is limited by its low molecular weight, dark color, and low solubility. In this work, an efficient crosslinking reaction with N,N-Dimethylformamide (DMF) and 1,6-dibromohexane was proposed for adjusting the molecular weight and color of lignin. The chemical structure of alkylation lignin was systematically investigated by gel permeation chromatography (GPC), ultraviolet spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and 2D heteronuclear single quantum correlation nuclear magnetic resonance (HSQC NMR) spectra. After the alkylation modification, the molecular weights of the lignin were increased to 1643%. The resinol (β-β), β-aryl ether (β-O-4), and phenylcoumaran (β-5) linkages were still the main types of the linkages. The formation of β-β linkage would be inhibited at high temperatures. The color reduction of lignin can be attributed to the low content of chromophores and low packing density. This alkylation lignin will be a new and general approach for developing molecular weight-controlled and light-colored lignins, which can find more applications in cosmetics, packing, and other fields.

New robustaflavone from Garcinia latissima Miq. leave and Its antibacterial activity.

Isolation and determination of antibacterial compounds from plants are essential to obtain a new antibacterial as a substitute for conventional resistant antibiotics. This study aims to isolate and identify a new robustaflavone as antibacterial activity from Garcinia latissima Miq. leave. In this study, the isolation process was carried out using column chromatography followed by preparative thin layer chromatography (TLC) based on the TLC profile. The fraction D was tested for anti-bacterial Bacillus subtilis using the TLC bioautography method. The isolates obtained were then identified using 1H-NMR, 13C-NMR, distortionless enhancement by polarization transfer, heteronuclear single quantum coherence, and heteronuclear multiple bond coherence. The Activity assay of the isolate was performed using the microdilution method. A pure compound obtained the result of the separation process with eluent n-hexane: Ethyl acetate (3:2) with Rf 0.6. This spot follows the spot in the contact bioautographic result of fraction D, the spot with Rf 0.6 gives an inhibition zone. After identifying and purifying the isolate were known as Robustaflavone, this compound has activity against B. subtilis with a (minimum inhibitory concentration) value of 2500 ppm. Robustaflavone successfully isolated and identified from G. latissima leave and its antibacterial activity.

1H-13C heteronuclear single quantum coherence NMR evidence for iodination of natural organic matter influencing organo-iodine mobility in the environment

The complex biogeochemical behavior of iodine (I) isotopes and their interaction with natural organic matter (NOM) pose a challenge for transport models. Here, we present results from iodination experiments with humic acid (HA) and fulvic acid (FA) using 1H-13C heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectroscopy. Even though not a quantitative approach, 1H-13C HSQC NMR corroborated that iodination of NOM occurs primarily through aromatic electrophilic substitution of proton by I, and also revealed how iodination chemically alters HA and FA in a manner that potentially affects the mobility of iodinated NOM in the environment. Three types of iodination experiments were conducted with HA and FA: a) non-enzymatic iodination by IO3− (pH 3) and I− (pH 4 and 7), b) addition of lactoperoxidase to promote I−-iodination in the presence of the co-substrate, H2O2 (pH 7), and c) addition of laccase for facilitating I−-iodination in the presence of O2, with or without a mediator (pH 4). When mediators or H2O2 were present, extracellular oxidases and peroxidases enhanced I− incorporation into NOM by between 54% and 3400%. Iodination of HA, which was less than that of FA, enhanced HA's stability (inferred from increases in aliphatic compounds, decreases in carbohydrate moieties, and thus increased molecular hydrophobicity) yet reduced HA's tendency to incorporate more iodine. As such, HA is expected to act more as a sink for iodine in the environment. In contrast, iodination of FA appeared to generate additional iodine binding sites, which resulted in greater iodine uptake capability and enhanced mobility (inferred from decreases in aliphatic compounds, increases in carbohydrates, and thus decreases in molecular hydrophobicity). These results indicate that certain NOM moieties may enhance while others may inhibit radioiodine mobility in the aqueous environment.

Structural characterization using 2D NMR spectroscopy and TMAH-GC × GC-MS: Application to humic acids from soils of an integrated agricultural system and an Atlantic native forest

Understanding the chemical make-up of soils and their structure is critical for constraining the role of soil organic matter (SOM) into the global biogeochemical cycles, as well as to understand the capability of SOM to sequester carbon and mitigate greenhouse gas emissions. Here, we use two-dimensional 1H-13C heteronuclear single quantum coherence nuclear magnetic resonance (2D 1H-13C HSQC NMR) spectroscopy to structurally characterize the most refractory component of SOM, the humic acid (HA). The observations from 2D 1H-13C HSQC NMR were coupled with lignin phenol and fatty acid measurements using tetramethylammonium hydroxide (TMAH) thermochemolysis – two-dimensional gas chromatography – mass spectrometry (TMAH-GC × GC-MS). We studied humic acids extracted from an integrated Crop – Livestock – Forest System (CLFS) agricultural area and an undisturbed Atlantic Native Forest (NF) area. We evaluated soils from two different depths: the topsoil (0–20 cm) and subsoil (60–100 cm) layers, and reveal the presence of oxidized ligninaceous phenols as we had previously hypothesized. Collectively, our results indicate that there are significant oxidative processes with increasing soil depth which are more pronounced in the CLFS relative to the NF area. Degradation of stearic acid with increasing depth in the CLFS soils indicated that the CLFS is more microbiologically active than NF. Therefore, CLFS is less biochemically stable than we originally perceived. The enhanced bio-reactivity of CLFS is likely driving the enhanced carbon sequestration in the CLFS soils. This is perhaps due to the diversity of biomass remnants available at the CLFS soil rhizosphere which allows for more different types of biomass to be sequestered as oxidized ligninaceous phenols. Our results employing structural characterization with 2D 1H-13C HSQC NMR and TMAH-GC × GC-MS provide a new layer of knowledge about the practice of integrated agricultural systems and allow us to understand the structure and fate of sequestered carbon in soils from different soil environments.

Chemical Structures of Adhesive and Interphase Parts in Sucrose/Citric Acid Type Adhesive Wood-Based Molding Derived from Japanese Cedar (Cryptomeria japonica).

In sucrose/citric acid based wood adhesive, the detailed bonding mechanism has still been unknown. Here, we investigated the detailed chemical structures of this adhesive wood (Japanese cedar)-based molding by using heteronuclear single quantum coherence–nuclear magnetic resonance (HSQC-NMR). NMR peaks associated with the furan-type structure appeared, suggesting that the furan compound was formed from sucrose and converted to a furan polymer during the adhesive process and that some of the furan structures in the polymers were ester-bonded with citric acid. The secondary forces between the furan polymers and wood components were thought to contribute to the adhesive effect. In our analysis of the interphase structure, primary hydroxyl groups of both polysaccharides and of lignin substructures were found to be esterified with citric acid. Additionally, some of the glycosidic bonds in polysaccharides were cleaved during the acidic condition produced by citric acid. The above results provided evidence of the polymerization of sucrose-derived 5-HMF, the esterification of wood components, and the degradation of polysaccharides during the molding process. Citric acid functioned as a clamp between the obtained furan polymer and the wood components. The sucrose/citric acid based wood adhesive can be defined as a hybrid-type wood adhesive, involving both secondary forces and chemical bonding interactions.

Boosting the resolution of multidimensional NMR spectra by complete removal of proton spin multiplicities

Over decades multidimensional NMR spectroscopy has become an indispensable tool for structure elucidation of natural products, peptides and medium sized to large proteins. Heteronuclear single quantum coherence (HSQC) spectroscopy is one of the work horses in that field often used to map structural connectivity between protons and carbons or other hetero nuclei. In overcrowded HSQC spectra, proton multiplet structures of cross peaks set a limit to the power of resolution and make a straightforward assignment difficult. In this work, we provide a solution to improve these penalties by completely removing the proton spin multiplet structure of HSQC cross peaks. Previously reported sideband artefacts are diminished leading to HSQC spectra with singlet responses for all types of proton multiplicities. For sideband suppression, the idea of restricted random delay (RRD) in chunk interrupted data acquisition is introduced and exemplified. The problem of irreducible residual doublet splitting of diastereotopic CH2 groups is simply solved by using a phase sensitive JRES approach in conjunction with echo processing and real time broadband homodecoupling (BBHD) HSQC, applied as a 3D experiment. Advantages and limitations of the method is presented and discussed.

Maleated soybean oil derivatives as versatile reactive diluents: Synthesis, characterization, and evaluation

AbstractFour maleated soybean oil (SBO) derivatives were synthesized and used as bio‐based reactive diluents (RDs) for alkyd coating systems. The RDs were prepared in a two‐step fashion: (a) Maleation of SBO with maleic anhydride (MA) via an ene reaction followed by (b) a nucleophilic acyl substitution of the grafted succinic anhydride with allyl‐ or methacrylate‐functional molecules. The structures were characterized using 1H NMR, 13C NMR, 13C‐1H heteronuclear single quantum coherence NMR, 13C‐1H heteronuclear multiple bond correlation NMR spectroscopy, FT‐IR (ATR) spectroscopy, and MALDI‐ToF‐MS. The 2D NMR spectroscopy techniques helped confirm that MA was successfully grafted onto SBO. FT‐IR spectroscopy identified new absorption bands at 1780 and 1850 cm−1, following the maleation reaction, corresponding to the anhydride functionality. Alkyd coating formulations were prepared with either 0, 10, 20, or 30 wt% of the different RDs. Brookfield viscosity measurements were conducted on the formulations to measure the efficiency of the SBO‐based RDs. At 20 wt% loading level, the RDs typically reduced the viscosity of the base alkyd resin by 50%–60%. Coatings tests and gel content measurements were also conducted in order to understand the effects that these RDs had on the performance of the alkyd coatings.

Suppression Effect of Gamma-Valerolactone on the Mild Alkaline Pretreatment of Hybrid Pennisetum

Gamma-valerolactone (GVL) assisted NaOH/water pretreatment for hybrid pennisetum delignification was first investigated. Contrary to the hypothesized results, the addition of GVL significantly suppressed the delignification and hemicellulose removal by mild alkaline pretreatment (37 °C for 24 h). An addition of GVL to GVL/NaOH molar ratio of 1/1 resulted in lignin removal at ∼23% compared with that at ∼75% without GVL. This suppression effect was further confirmed by scanning electron microscopy and Fourier-transform infrared spectroscopy. The extracted lignin was further characterized by two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance spectroscopy, suggesting that the addition of GVL suppressed the decomposition of large lignin molecules by NaOH/water and the release of hemicellulose, resulting in extracted lignin with lower molecular weights and less hemicellulose. The decrease in the partial alkalinity (PA) of alkaline solutions caused by GVL addition was likely the main reason for the suppressed delignification, suggested by the good correlation between PA and delignification in both GVL/NaOH/water and NaOH/water (R2 = 0.967). This study suggested that solvent-assisted alkaline pretreatment can also result in an unwanted suppression effect, and the changes in PA could be referred to as an indicator while selecting organic solvents.

Comparison of organic acid-based organosolv lignins extracted from the residues of five annual crops

Organosolv lignins were extracted from corn stover, wheat, rice straw, reed straw, and sugarcane bagasse using a mixture of acetic and formic acids, at relatively low temperature and atmospheric pressure. Lignin content, residual carbohydrates, ash levels, proteins, and molecular weights were determined in each extracted lignin. The lignin content of all samples was relatively high, confirming the performance of the pretreatment process. The low molecular weights were in a narrow range, in accordance with the organosolv lignin molar masses. However, some differences between studied lignins were highlighted, in particular in rice straw lignin, which contained the highest silica, calcium, and nitrogen contents. Nuclear magnetic resonance spectroscopies (31P and semi-quantitative Heteronuclear Single Quantum Correlation) underlined the structural similarities and differences between these organosolv lignins. Corn stover and sugarcane bagasse lignins were rich in non-methoxylated (H-Unit) or mono-methoxylated (G-Unit) phenolic units, making them the best promising candidates for production of phenolic resins. Wheat straw lignin was richer in aliphatic OH than in phenolic OH. This is an advantage for use as polyol substitute in polyurethane synthesis. Reed straw lignin was less specific, with a balanced content of OH groups. However, it contained a high concentration of β-O-4 linkages, which is favorable for depolymerization.

Demethylation, phenolation, and depolymerization of lignin for the synthesis of lignin-based epoxy resin via a one-pot strategy

A one-pot method to synthesize demethylated-phenolated lignin (DPL) and demethylated-phenolated lignin-based epoxy resin (DPLINP) using phenol as a phenolic reagent and polymerization inhibitor was proposed in this study. This method avoids the occurrence of polycondensation caused by demethylation and simplifies the synthesis of lignin-based epoxy resins. Phenolated lignin (PL)-based epoxy resin (PLINP) and demethylated lignin (DL)-based epoxy resin (DLINP) were prepared to investigate the effects of different modification methods on the resulting lignin and epoxy resin. The results of 1H nuclear magnetic resonance (NMR), Fourier transform infrared, and gel permeation chromatography analyses indicated that the introduction of phenol could significantly reduce the occurrence of polycondensation while also decreasing the molecular weight and increasing the content of phenolic hydroxyl in DPL compared with DL and PL. 2D heteronuclear single quantum coherence NMR analysis was used to further investigate the structural changes of lignin during the modification process. The results obtained via dynamic mechanical analysis, flexural test, and impact test demonstrated that demethylated-phenolated lignin-based epoxy network (DPLIEN) possessed an improved cross-linking density, glass transition temperature, and mechanical performance compared with all other epoxy resins. Moreover, the polycondensation of DL caused by demethylation deteriorated the mechanical performance of demethylated lignin-based epoxy network (DLIEN). The thermal stability of DPLIEN was decreased compared with phenolated lignin-based epoxy network (PLIEN), as the removal of methoxy groups may have reduced the protective char layer formation in DPLIEN. This study provides a novel method for the modification of lignin and synthesis of lignin-based epoxy resins.

Composition and Protein Precipitation Capacity of Condensed Tannins in Purple Prairie Clover (Dalea purpurea Vent.).

This study aimed to determine the concentration and composition of condensed tannins (CT) in different tissues of purple prairie clover (PPC; Dalea purpurea Vent.) at different maturities and to determine their protein-precipitating capacity. The compositions of CT were elucidated after thiolysis with benzyl mercaptan followed by high-performance liquid-chromatography (HPLC) and 1H–13C heteronuclear single quantum coherence (HSQC) NMR spectroscopy. The results indicated that PPC flowering heads contained the highest CT concentration. Purple prairie clover CT consisted mainly of epicatechin (EC) and epigallocatechin (EGC) subunits. CT in the leaves were composed of more EC and less EGC than CT in stems and flowering heads at both the early flowering (EF) and late flowering (LF) head stages. The mean degree of polymerization was the highest for CT in stems and increased with maturity. CT isolated from PPC leaves at the early flowering head stage exhibited the greatest biological activity in terms of protein precipitation. Overall, the CT in PPC were predominantly procyanidins and the concentration and composition varied among the plant tissues and with maturity.

Probing multiple enzymatic methylation events in real time with NMR spectroscopy

Post-translational modification (PTM) of proteins is of critical importance to the regulation of many cellular processes in eukaryotic organisms. One of the most well-studied protein PTMs is methylation, wherein an enzyme catalyzes the transfer of a methyl group from a cofactor to a lysine or arginine side chain. Lysine methylation is especially abundant in the histone tails and is an important marker for denoting active or repressed genes. Given their relevance to transcriptional regulation, the study of methyltransferase function through in vitro experiments is an important stepping stone toward understanding the complex mechanisms of regulated gene expression. To date, most methyltransferase characterization strategies rely on the use of radioactive cofactors, detection of a methyl transfer byproduct, or discontinuous-type assays. Although such methods are suitable for some applications, information about multiple methylation events and kinetic intermediates is often lost. Herein, we describe the use of two-dimensional NMR to monitor mono-, di-, and trimethylation in a single reaction tube. To do so, we incorporated 13C into the donor methyl group of the enzyme cofactor S-adenosyl methionine. In this way, we may study enzymatic methylation by monitoring the appearance of distinct resonances corresponding to mono-, di-, or trimethyl lysine without the need to isotopically enrich the substrate. To demonstrate the capabilities of this method, we evaluated the activity of three lysine methyltransferases, Set7, MWRAD2 (MLL1 complex), and PRDM9, toward the histone H3 tail. We monitored mono- or multimethylation of histone H3 tail at lysine 4 through sequential short two-dimensional heteronuclear single quantum coherence experiments and fit the resulting progress curves to first-order kinetic models. In summary, NMR detection of PTMs in one-pot, real-time reaction using facile cofactor isotopic enrichment shows promise as a method toward understanding the intricate mechanisms of methyltransferases and other enzymes.

Elucidation of ligninolysis mechanism of a newly isolated white-rot basidiomycete Trametes hirsuta X-13

BackgroundLignin is a complex aromatic heteropolymer comprising 15–30% dry weight of the lignocellulose. The complex structural characteristic of lignin renders it difficult for value-added utilization. Exploring efficient lignin-degrading microorganisms and investigating their lignin-degradation mechanisms would be beneficial for promoting lignin valorization. In this study, a newly isolated white-rot basidiomycete, Trametes hirsuta X-13, with capacity to utilize alkaline lignin as the sole substrate was investigated.ResultsThe analysis of the fermentation properties of T. hirsuta X-13 using alkaline lignin as the sole substrate, including the mycelial growth, activities of ligninolytic enzymes and the rates of lignin degradation and decolorization confirmed its great ligninolysis capacity. The maximum lignin degradation rate reached 39.8% after 11 days of T. hirsuta X-13 treatment, which was higher than that of reported fungi under the same condition. Fourier transform infrared spectrometry (FTIR), gas chromatography–mass spectrometry (GC–MS) scanning electron micrographs (SEM), two-dimensional heteronuclear single quantum coherence NMR analysis (2D-HSQC NMR) collaborated with pyrolysis gas chromatography–mass spectrometry (py-GC/MS) analyses proved that lignin structure was severely deconstructed along with amounts of monomer aromatics generated. Furthermore, according to those chemical analysis, in addition to canonical Cα–Cβ breakage, the cleavage of lignin interunit linkages of β–β might also occur by T. hirsuta X-13.ConclusionsThis study characterized a newly isolated white-rot basidiomycete T. hirsuta X-13 with impressive alkaline lignin degradation ability and provided mechanistic insight into its ligninolysis mechanism, which will be valuable for the development of lignin valorization strategies.

Catalytic Hydrogenolysis of Lignin: The Influence of Minor Units and Saccharides.

The precise elucidation of native lignin structures plays a vital role for the development of “lignin first” strategies such as reductive catalytic fractionation. The structure of lignin and composition of the starting material has a major impact on the product yield and distribution. Here, the differences in structure of lignin from birch, pine, reed, and walnut shell were investigated by combining detailed analysis of the whole cell wall material, residual enzyme lignin, and milled wood lignin. The results of the 2D heteronuclear single quantum coherence NMR analysis could be correlated to the product from Ru/C‐catalyzed hydrogenolysis if monomeric products from ferulate and p‐coumaryl and its analogous units were also appropriately considered. Notably, residual polysaccharide constituents seemed to influence the selectivity towards hydroxy‐containing monomers. The results reinforced the importance of adequate structural characterization and compositional analysis of the starting materials as well as distinct (dis)advantages of specific types of structural characterization and isolation methods for guiding valorization potential of different biomass feedstocks.

Lignin Resists High-Intensity Electron Beam Irradiation.

The electron beam irradiation (EBI) of native lignin has received little attention. Thus, its potential use in lignin-based biorefineries is not fully understood. EBI was applied to selected lignin samples and the structural and chemical changes were analyzed, revealing the suitability, limitations, and potential purpose of EBI in wood biorefineries. Isolated milled wood, kraft, and sulfite lignin from beech and eucalyptus were subjected to up to 200 kGy of irradiation. The analysis included gel permeation chromatography for molar masses, heteronuclear single quantum coherence (HSQC)- and 31P NMR and headspace gas chromatography-mass spectrometry for functional groups, and thermogravimetric analysis for thermal stability. Most samples resisted irradiation. Subtle changes occurred in the molecular weight distribution and thermal stability of milled wood lignin. EBI was found to be a suitable pretreatment method for woody biomass if the avoidance of lignin condensation and chemical modification is a high priority.

A comprehensive study of the promoting effect of manganese on white rot fungal treatment for enzymatic hydrolysis of woody and grass lignocellulose

BackgroundThe efficiency of biological systems as an option for pretreating lignocellulosic biomass has to be improved to make the process practical. Fungal treatment with manganese (Mn) addition for improving lignocellulosic biomass fractionation and enzyme accessibility were investigated in this study. The broad-spectrum effect was tested on two different types of feedstocks with three fungal species. Since the physicochemical and structural properties of biomass were the main changes caused by fungal degradation, detailed characterization of biomass structural features was conducted to understand the mechanism of Mn-enhanced biomass saccharification.ResultsThe glucose yields of fungal-treated poplar and wheat straw increased by 2.97- and 5.71-fold, respectively, after Mn addition. Particularly, over 90% of glucose yield was achieved in Mn-assisted Pleurotus ostreatus-treated wheat straw. A comparison study using pyrolysis gas chromatography mass spectrometry (Py-GC/MS) and two-dimensional 1H–13C heteronuclear single quantum coherence (2D HSQC) nuclear magnetic resonance (NMR) spectroscopy was conducted to elucidate the role of Mn addition on fungal disruption of the cross-linked structure of whole plant cell wall. The increased Cα-oxidized products was consistent with the enhanced cleavage of the major β-O-4 ether linkages in poplar and wheat straw lignin or in the wheat straw lignin–carbohydrate complexes (LCCs), which led to the reduced condensation degree in lignin and decreased lignin content in Mn-assisted fungal-treated biomass. The correlation analysis and principal component analysis (PCA) further demonstrated that Mn addition to fungal treatment enhanced bond cleavage in lignin, especially the β-O-4 ether linkage cleavage played the dominant role in removing the biomass recalcitrance and contributing to the glucose yield enhancement. Meanwhile, enhanced deconstruction of LCCs was important in reducing wheat straw recalcitrance. The findings provided not only mechanistic insights into the Mn-enhanced biomass digestibility by fungus, but also a strategy for improving biological pretreatment efficiency of lignocellulose.ConclusionThe mechanism of enhanced saccharification of biomass by Mn-assisted fungal treatment mainly through Cα-oxidative cleavage of β-O-4 ether linkages further led to the decreased condensation degree in lignin, as a result, biomass recalcitrance was significantly reduced by Mn addition.Graphic abstract