Structural Changes Of Lignin Research Articles

Surface Morphology and Chemical Changes of Maple and Beech Cut Through by CO2 Laser Under Different Angles Relative to the Wood Grain

This paper examined the surface morphology of maple and beech cut through by CO2 laser under different angles relative to the wood grain: 0°, 15°, 30°, 45°, 60°, 75°, and 90°. In the analysis, stylus measurements, stereo-microscopic images, and chemical changes were considered. Laser uncovers more wood anatomical details, with enhanced clarity, when the cutting transitions from along the grain to across the grain. This is particularly noticeable in the earlywood and is more pronounced in maple compared to beech. The first tissue of earlywood was deeply ablated by the laser, leading to a wavy anatomical pattern, which is more visible for higher angles of laser cutting in relation to the wood grain. The anatomical structure of beech was more affected by carbonization in comparison to maple and had a significantly higher core roughness, Rk. For both species, the worst surface roughness occurred when cutting at 15°. In maple, the laser caused more degradation of the polysaccharides compared to beech, and this impact was particularly noticeable parallel to the grain rather than at a 90° angle. The degradation of hemicelluloses occurred in parallel with more advanced cellulose degradation for beech compared to maple and for cutting along the grain compared to across the grain. Structural changes in lignin, such as condensation processes, were observed for both species.

Effects of alkali-soluble hemicellulose separation on the structure of bamboo lignin and lignin-carbohydrate complexes

In this study, alkali treatment was used to preferentially separate hemicellulose, and the structural changes of lignin and lignin-carbohydrate complexes (LCC) in the residual bamboo solids were investigated. The chemical composition and structural characteristics of lignin and LCC were analyzed. After alkali treatment, the monosaccharide content in bamboo lignin decreased from 7.62 % to 2.90 % compared to the raw material. The obtained lignin exhibited higher purity and larger molecular weight, with the macromolecular lignin linked by β-O-4′ bonds remaining almost undegraded. After alkali treatment, the linkage bonds in the Björkman LCC were composed of phenyl glycoside (PhGlc), and the bond in LCC-AcOH consisted of benzyl ether (BE). The results showed that the preferential extraction of hemicellulose preserved the biopolymer structure of lignin in bamboo, thereby providing an important basis for the comprehensive separation and utilization of bamboo components.

Quantification of Native Lignin Structural Features with Gel‐Phase 2D‐HSQC0 Reveals Lignin Structural Changes During Extraction

AbstractOur ability to study and valorize the lignin fraction of biomass is hampered by the fundamental and still unmet challenge of precisely quantifying native lignin's structural features. Here, we developed a rapid elevated‐temperature 1H−13C Heteronuclear Single‐Quantum Coherence Zero (HSQC0) NMR method that enables this precise quantification of native lignin structural characteristics even with whole plant cell wall (WPCW) NMR spectroscopy, overcoming fast spin relaxation in the gel phase. We also formulated a Gaussian fitting algorithm to perform automatic and reliable spectral integration. By combining HSQC0 measurements with yield measurements following depolymerisation, we can confirm the combinatorial nature of radical coupling reactions during biosynthesis leading to a random sequential organization of linkages within a largely linear lignin chain. Such analyses illustrate how this analytical method can greatly facilitate the study of native lignin structure, which can then be used for fundamental studies or to understand lignin depolymerization methods like reductive catalytic fractionation or aldehyde‐assisted fractionation.

Quantification of Native Lignin Structural Features with Gel-Phase 2D-HSQC0 Reveals Lignin Structural Changes During Extraction.

Our ability to study and valorize the lignin fraction of biomass is hampered by the fundamental and still unmet challenge of precisely quantifying native lignin's structural features. Here, we developed a rapid elevated-temperature 1H-13C Heteronuclear Single-Quantum Coherence Zero (HSQC0) NMR method that enables this precise quantification of native lignin structural characteristics even with whole plant cell wall (WPCW) NMR spectroscopy, overcoming fast spin relaxation in the gel phase. We also formulated a Gaussian fitting algorithm to perform automatic and reliable spectral integration. By combining HSQC0 measurements with yield measurements following depolymerisation, we can confirm the combinatorial nature of radical coupling reactions during biosynthesis leading to a random sequential organization of linkages within a largely linear lignin chain. Such analyses illustrate how this analytical method can greatly facilitate the study of native lignin structure, which can then be used for fundamental studies or to understand lignin depolymerization methods like reductive catalytic fractionation or aldehyde-assisted fractionation.

Mechanism Studies in Electrochemical Depolymerization Processes of Alkali and Organosolv Lignin

AbstractTo better understand the process and mechanism of lignin structural changes and relative product formation during PbO2 anode electrolysis, a Ti/Sb−SnO2/PbO2 composite electrode has been prepared and utilized for the alkaline electrolytic oxidation of both alkali lignin (AL) and bagasse organosolv lignin (OL). By combining electrochemical measurements, in‐situ FTIR, 2D‐HSQC NMR, and product distribution analysis, we not only investigate the impact of influence factors on the process of lignin depolymerization, but also reveal the depolymerization mechanism from the whole process of lignin structural changes in details. Under certain electrolysis conditions, the highest vanillin yield, 10.1 mg g−1, has been obtained through the electrolysis of alkali lignin, whereas the highest syringaldehyde yield of 4.0 mg g−1 has been achieved during the electrolysis of organic solvent lignin. In particular, AL mainly performs depolymerization through the selective cleavage C−C bonds to produce a series of G unit monomer products of vanillin, acetovanillone, and vanillic acid. By contrast, the depolymerization of OL shows significant selectivity in H units. Particularly, the production of the major H unit monomer product of 4‐vinylphenol has been confirmed by IR spectroelectrochemical studies from the oxidation and decarboxylation of p‐coumaric acid unit.

Solubilization and structural changes of lignin in naked oat stems during subcritical water autohydrolysis

In this study, the solubilization and structural changes of lignin in naked oat stems were investigated under subcritical water autohydrolysis systems (170–210 °C, 0.68–1.85 MPa). In this system, Hemicellulose was preferentially hydrolyzed in the liquid water at elevated temperatures, leading to the production of acetic acid and glucuronic acid, which acidified the reaction system. Under acidic and high-temperature conditions, lignin primarily underwent degradation and condensation reactions. At autohydrolysis temperatures below 190 °C and autohydrolysis pressures below 1.22 MPa, lignin degradation was predominant, realizing a maximum lignin removal of 47.8 % and breakage of numerous β-O-4 bonds from lignin. At autohydrolysis temperatures above 190 °C and autohydrolysis pressures above 1.22 MPa, lignin condensation dominated, with an increase in the amount of organic acids generated upon hemicellulose degradation, leading to condensation reactions with the degraded low-molecular-weight lignin. The degree of lignin condensation was positively correlated with the temperature of the reaction system. This study provides essential insights into the dynamic changes in the structure of lignin in both the hydrolysis residue and hydrolysis solution during subcritical water autohydrolysis.

Revealing the structural changes of lignin in Chinese quince (Chaenomeles sinensis) fruit as it matures

Chinese quince fruits (Chaenomeles sinensis) contain substantial amounts of lignin; however, the exact structure of lignin remains to be investigated. In this study, milled wood lignins (Milled wood lignin (MWL)-1, MWL-2, MWL-3, MWL-4, MWL-5, and MWL-6) were extracted from fruits harvested once a month from May to October 2019 to investigate their structural evolution during fruit growth. The samples were characterized via High-performance anion exchange chromatography (HPAEC), Fourier transform–infrared spectroscopy (FT-IR), gel permeation chromatography (GPC), thermogravimetric (TGA), pyrolysis–gas chromatography/mass spectrometry (Py–GC/MS) and NMR (2D-heteronuclear single quantum coherence (HSQC) and 31P). The MWL samples in all fruit growth stages were GS-type lignin and lignin core undergoing minimal alterations during fruit development. The predominant linkage in the lignin structure was β-O-4′, followed by β-β' and β-5′. Galactose and glucose were the main monosaccharides associated with MWL. In MWL-6, the lignin exhibited the highest homogeneity and thermal stability. As the fruit matured, a gradual increase in the β-O-4′ proportion and the ratio of S/G was observed. The results provide comprehensive characterization of the cell wall lignin of quince fruit as it matures. This study could inspire innovative applications of quince fruit lignin and provide the optimal harvest time for lignin utilization.

Kinetic understanding of fiber surface lignin effects on cellulase adsorption and hydrolysis

Cellulase hydrolysis is strongly affected by adsorption, which is inhibited by the non-productive binding between cellulase and lignin. However, lignin-bonded cellulase has also been found to be active in hydrolysis. Thus, in this work, lignin effects on cellulase adsorption and hydrolysis were studied using fibers with various surface lignin contents and structures based on kinetics. The surface lignin was changed by mechanical refining. Cellulase adsorbed on lignin showed reactivity to hydrolyze cellulose nearby according to the kinetic results, which is, however, much weaker compared with that by cellulose-bonded cellulase. With the decrease of surface lignin, this reactivity is also decreasing. Both cellulase adsorption and hydrolysis were increasing at first and then decreasing with the decrease of surface lignin as well as the lignin re-adsorption. It was concluded that lignin plays an important role in cellulase adsorption and non-productive binding occurs more easily between cellulase and the re-adsorbed lignin, as well as the residual lignin. At a relatively high surface lignin level, lignin structural and distribution changes were found to have a better effect on cellulase hydrolysis compared with lignin removal.

LIGNOCELLULOSE BIOMASS DELIGNIFICATION USING ACID HYDROTROPE AS GREEN SOLVENT: A MINI-REVIEW

"Efficient and cost-effective conversion of lignocellulosic biomass into usable forms of energy presents unique challenges. Lignocellulosic biomass, comprising cellulose, hemicelluloses, and lignin, necessitates advanced conversion technologies. Common commercial delignification techniques, including kraft pulping, sulfite pulping, acid hydrolysis, and organosolv pulping, often involve harsh conditions leading to structural changes in lignin and environmental impacts. To address these issues, acid hydrotropes have emerged as a promising method for lignin extraction. Acid hydrotropes, represented by p-toluenesulfonic acid (p-TsOH), enable the solubilization of hydrophobic substances like lignin. This mini-review provides an overview of various lignocellulose fractionation techniques and explores the acid hydrotrope approach. The mechanism behind acid hydrotropic fractionation is discussed, and its performance is evaluated. In conclusion, the review emphasizes the pivotal role of the acid hydrotrope approach in advancing lignocellulosic biomass conversion technology, promoting a sustainable and efficient bio-based economy."

Production and characterization of novel thermostable CotA-laccase from Bacillus altitudinis SL7 and its application for lignin degradation

Laccases are multi-copper oxidases and found in ligninolytic bacteria catalyzing the oxidation of both phenolic and non-phenolic compounds, however its application in lignin degradation suffers due to low oxidation rate, which have intensified the search for new laccases. In the present study, spore coat A protein (CotA) encoding gene having laccase like activity from Bacillus altitudinis SL7 (CotA-SL7) was cloned and expressed in Escherichia coli. The purified CotA-SL7 was active at wide range of temperature and pH with optimum activity at 55 °C and pH 5.0. The kinetic parameters of CotA-SL7 was determined with Km, Vmax, and kcat values 0.4 mM, 2777 μmol/min/mg, and 5194 s−1, respectively. Molecular docking revealed the presence of Pro, Phe, Asp, Asn, His, and Ile residues at the active site taking part in the oxidation of ABTS. The purified CotA-SL7 reduced lignin contents by 31 % and changes in lignin structure were analyzed through fourier transformed infrared spectroscopy (FTIR), scanning electron microsscopy (SEM) and gas chromatography mass-spectrometry (GC-MS). The appearance of low molecular size compounds clearly indicates the cleavage of lignin polymer and opening of the benzene ring by purified CotA-SL7. Thus, high catalytic efficiency of CotA-SL7 makes it a suitable bio-catalyst for remediation of lignin contaminated wastewater from pulp and paper industries with clear insights into lignin degradation at molecular level.

Fiber Melt Spinning and Thermo-Stabilization of Para-Rubber Wood Lignin: An Approach for Fully Biomass Precursor Preparation.

Para-rubber wood (PRW) lignin, extracted from agricultural waste, was successfully melt-spun to fibers and thermo-stabilized without employing auxiliary additives. 31P NMR analysis revealed that PRW-lignin contained mainly a syringyl unit of phenolic C5-substituted OH group, which enabled melt flow during fiber spinning, as well as a guaiacyl unit which offered the ability to cross-link during thermo-stabilization. Thermo-stabilized fibers with no fusion were achieved at 250 °C with the heating rate of 0.1 °C/min. Structural changes in the fibers during stabilization were systematically investigated using FTIR and XPS analyses. From the results, changes in the intensities of characteristic bands relating to C-H stretching, aromatic C-H stretching, and C=O stretching indicated structural changes of lignin toward aromaticity via oxidation reactions. XPS analysis of the fibers carbonized at 900, 1000, and 1200 °C revealed an increase in carbon content from 72 to 87 wt %. and a decrease in oxygen content from 28 to 13 wt %. with the increasing carbonization temperature. The weight loss of carbonized fibers was in the range of 73.6 to 88.7%. The high weight loss of fibers carbonized at 1200 °C was explained partly due to the thermal decomposition of disordered carbon. The tensile strength and modulus of carbonized fibers were 163.0 and 275.1 MPa, respectively. This study demonstrates an approach to prepare a fully biomass precursor fiber and contributes to the exploration of the potential use of lignin from biomass waste.

Modification of a Marine Pine Kraft Lignin Sample by Enzymatic Treatment with a Pycnoporus cinnabarinus Laccase.

Here, we report work on developing an enzymatic process to improve the functionalities of industrial lignin. A kraft lignin sample prepared from marine pine was treated with the high-redox-potential laccase from the basidiomycete fungus Pycnoporus cinnabarinus at three different concentrations and pH conditions, and with and without the chemical mediator 1-hydroxybenzotriazole (HBT). Laccase activity was tested in the presence and absence of kraft lignin. The optimum pH of PciLac was initially 4.0 in the presence and absence of lignin, but at incubation times over 6 h, higher activities were found at pH 4.5 in the presence of lignin. Structural changes in lignin were investigated by Fourier-transform infrared spectroscopy (FTIR) with differential scanning calorimetry (DSC), and solvent-extractable fractions were analyzed using high-performance size-exclusion chromatography (HPSEC) and gas chromatography-mass spectrometry (GC-MS). The FTIR spectral data were analyzed with two successive multivariate series using principal component analysis (PCA) and ANOVA statistical analysis to identify the best conditions for the largest range of chemical modifications. DSC combined with modulated DSC (MDSC) revealed that the greatest effect on glass transition temperature (Tg) was obtained at 130 U g cm-1 and pH 4.5, with the laccase alone or combined with HBT. HPSEC data suggested that the laccase treatments led to concomitant phenomena of oligomerization and depolymerization, and GC-MS revealed that the reactivity of the extractable phenolic monomers depended on the conditions tested. This study demonstrates that P. cinnabarinus laccase can be used to modify marine pine kraft lignin, and that the set of analytical methods implemented here provides a valuable tool for screening enzymatic treatment conditions.

Green Fractionation and Structural Characterization of Lignin Nanoparticles via Carboxylic-Acid-Based Deep Eutectic Solvent (DES) Pretreatment

Green fractionation and a comprehensive overview of lignin molecular structures during the DES (deep eutectic solvent) pretreatment are very important for lignin valorization and the whole biorefinery process. Herein, intractable woody biomass (poplar wood) was pretreated with five types of carboxylic-acid-based DESs (acetamide served as an HBA (hydrogen bond acceptor), propanedioic acid, tartaric acid, malic acid, glutaric acid, and succinic acid served as HBDs (hydrogen bond donors)) under the optimized pretreatment conditions. Results showed that the optimal delignification ratio was achieved for tartaric-acid-based DES at 140 °C for 20 min under microwave-assisted heating. Two-dimensional HSQC NMR data demonstrated that the isolated poplar DES lignin consisted mainly of β-β, β-O-4 (normal and acylated forms), β-5, and esterified p-hydroxybenzoates (PBs) in different contents. Especially, the contents of β-O-4 in the isolated DES lignin fractions varied based on the pretreatment temperature and different chemical compositions of the DES. The antioxidant activity (2,2-diphenyl-1-picrylhydrazyl, DPPH analysis), microstructure (scanning electron microscope, SEM), and molecular weights (gel permeation chromatography, GPC) of the DES lignin fractions demonstrated that the DES delignification promoted the rapid assembly of lignin nanoparticles (LNPs) and could yield homogeneous lignin (1.23 < PDI < 1.58) with controlled nanometer size (30–170 nm) and good antioxidant activity. This study will improve the knowledge of structural changes of lignin during the different carboxylic-acid-based DES pretreatments and maximize the lignin valorization.

Revealing the structure and distribution changes of corn stalk lignin during the Organosolv Pretreatment

During the pretreatment of biomass, the research on the migration process of lignin in the cell wall and the change of chemical structure is very important for improving pretreatment and subsequent lignin upgrading. Hence, corn stalk lignin was extracted by 1, 4-dioxane organosolv system under mild conditions with varying pretreatment time and 0.1 M hydrochloric acid as catalyst. The heterogeneity of lignin removal in the different morphological areas of the cell wall was revealed by advanced imaging techniques. Results indicated that during the pretreatment with dioxane organosolv, the removal of lignin started from the secondary wall of the cell wall, and gradually developed outward to the compound middle lamella. Meanwhile, the resulting lignin obtained through different pretreatment times was characterized by two-dimensional heteronuclear single quantum coherence spectroscopy (2D-HSQC) and gel permeation chromatography (GPC). These characterization results indicated that there were more syringyl (S) units in the lignin that were removed first, and the proportion of guaiacyl (G) units was increased as the pretreatment continued. All the evidence supports the idea that the main reason for the significant differences in lignin structures at various periods of the lignin removal process is that the lignin come from different cell wall regions.

Effect of pretreatment severity on the inhibitory behaviors of larch lignins in enzymatic hydrolysis

To explore the underlying reasons for the decreased enzymatic hydrolysis yields of the acid pretreated softwood under the higher pretreatment severity, the residual bulk lignins, as one type lignin fraction with stronger inhibition, were isolated from larch (softwood) pretreated with different pretreatment severity in this study. The lignin structure changes during the pretreatment were comprehensively investigated by nuclear magnetic resonance (NMR) analysis. The larch lignins addition decreased the enzymatic hydrolysis of Avicel from 68.43% to 25.9–39.3%. The pretreatment intensified the inhibition of isolated lignins on enzymatic hydrolysis of Avicel. Furthermore, quartz crystal microbalance (QCM) analysis was applied to investigate the interaction between enzyme and lignins, distinguishing the reversible and irreversible enzyme adsorption. It demonstrated that the obviously higher irreversible adsorption of enzyme was observed on lignins from pretreated substrates with the higher severity. Additionally, the correlation between structure and inhibition of residual lignins in the enzymatic hydrolysis was then investigated with Pearson’s correlation heatmap. The findings can be used to develop theoretical guidelines for alleviating the residual lignin inhibition in the enzymatic hydrolysis of acidic pretreated softwood.

Valorization of lignin to obtain platform chemicals via bio‐electrochemical systems: batch and fed‐batch mode analysis

AbstractBACKGROUNDThere is tremendous potential for reusing lignin, which is generally discarded as waste. This research analyses the batch and fed‐batch mode of the microbial peroxide‐producing cell (MPPC), a type of bio‐electrochemical cell that produces H2O2, a potent green oxidant utilized in the oxidation of Kraft lignin to produce platform chemicals with simultaneous wastewater treatment.RESULTSThe batch mode MPPC had a total shelf life of 10 days, with a peak phase of 5 days and maximum H2O2 of 9.7 ± 0.06 mmol L−1, and voltage of 164 ± 0.021 mV. The intermittent nutrition feeding strategy of fed‐batch MPPCs aided microbe metabolic reactions, extending the system's lifespan to 21 days with a maximum voltage of 294 ± 0.01 to 525 ± 0.008 mV and H2O2 of 14.14 ± 0.12 to 32.96 ± 1.35 mmol L−1, respectively. This offered more exposure time for lignin hydroxyl radical oxidation by H2O2, with enhanced depolymerization of 37–80% of high lignin concentration in repeated cycles compared to batch mode, which accomplished only 53.46%. Fourier transform infrared spectroscopy confirmed the structural changes in lignin of all systems, displaying loss and disorientation of major functional groups with greater intensity in fed batch‐operated MPPCs. Platform chemicals with high commercial value, including guaiacol, ferulic acid, vanillin, and others, were identified using liquid chromatography–mass spectrometry. In terms of wastewater treatment, biochemical oxygen demand and chemical oxygen demand removal efficiency ranged from 59% to 83%, with fed‐batch being the most efficient.CONCLUSIONThis research suggests that fed‐batch mode MPPC is significantly more productive than batch mode MPPC for lignin valorization to produce platform chemicals, making it more sustainable, economical, and environmentally friendly. © 2023 Society of Chemical Industry (SCI).

Degradation of Alkaline Lignin in the Lactic Acid-Choline Chloride System under Mild Conditions

Lignin is a natural polymer, second only to cellulose in natural reserves. Degradation is one of the ways to achieve the high-value transformation of lignin. Deep eutectic solvent (DES) thermal degradation of lignin can be used as an excellent green degradation method. This paper introduces the degradation mechanism and effect of the lactic acid-choline chloride DES system in dissolving and degrading alkaline lignin, and the final solvent recovery. It can also be found from the scanning electron microscope (SEM) images that the surface of the degraded solid product is transformed from smooth to disordered. Fourier transform infrared (FTIR) spectroscopy and 1 H-NMR spectroscopy were used to characterize the changes in lignin functional groups during DES treatment. The results showed that the content of phenolic hydroxyl groups increased after degradation, indicating that the β-O-4 ether bond was broken. The molecular weight of the degraded lignin was observed by gel permeation chromatography (GPC), and the lignin residue with low molecular weight and narrow polydispersity index was obtained. The lowest average molecular weight (Mw) reached 2512 g/mol. The ratio of oxygen to carbon atoms in lignin increased substantially during degradation as measured by X-ray photoelectron spectroscopy (XPS), probably because DES treatment was accompanied by many oxidation reactions, which led to significant structural changes in lignin and a large number of ether bond breakage reactions during the reaction. The main final degradation products are aromatic monomers, vanillin, butyrovanillone, etc.

Poplar lignin structural changes during extraction in γ-valerolactone (GVL)

In this paper, we describe an approach for producing both high quality and high quantity of lignin by studying structural change of lignin during treatment of poplar wood in γ-valerolactone (GVL) for a range of temperatures (80–120 °C) and reaction time at temperature (1–24 h).

Evaluation of oxy-organosolv pretreatment on lignin extraction from wheat straw

To develop a characteristic “Lignin-first” strategy, the oxy-organosolv delignification processes under mild conditions were comprehensively investigated. Results showed that lignin yield could achieve about 50 % under the optimum process conditions of ethanol concentration 80 %, temperature 90 °C, liquid to wheat straw ratio 25:1 for powdery-scale substrates, which was 65.0 % higher than that for rod-scale substrates under the same conditions. The lignin structural and carbohydrate component results demonstrated the employment of oxygen induced great quantities of lignin dissolving out on the premise of little carbohydrate component (<1 %) and lignin structural (mainly β-O-4 units) changes. Moreover, based on the molecular weight and polydiversity comparison results, the aqueous oxygen could transfer homogeneously in mild organosolv system and result in lignin degradation uniformly. Besides, the employment of oxygen assisted in not only extending the massive lignin removal stage to 30 min and 50 min for P-OEEL and R-OEEL respectively, but also boost the delignification rate with comparison to P-EL and R-EL. Lastly, the excellent anti-oxidant properties of lignin from oxy-organosolv process were demonstrated by scavenging DPPH and ABTS radicals. The economic calculations showed that the cost for lignin production were about 1.58USD/g lignin from powdery-scale wheat straw, providing a competitive route for high-value utilize waste biomass.

Ligninolytic characteristics of Pleurotus ostreatus cultivated in cotton stalk media.

Biodelignification is widely regarded as a low-efficiency process because it is usually slow and difficult to control. To improve its efficiency and understand its mechanism, the present study analyzed the delignification characteristics of Pleurotus ostreatus grown on a cotton stalk medium. The results demonstrated that all strains of P. ostreatus can selectively degrade the cotton stalk lignin. When cultured in a cotton stalk medium for 60 days, P. ostreatus degraded lignin primarily during its mycelium growth with up to 54.04% lignin degradation and produced laccase and manganese dependent peroxidase with high activity levels at the peaks of 70.17 U/ml and 62.39 U/ml, respectively, but no detectable lignin peroxidase. The results of nuclear magnetic resonance spectroscopy and Fourier transform infrared spectroscopy analyses of significant changes in lignin structure revealed that syringyl (S) lignin units were more degraded than guaiacyl (G) lignin units, with a significantly elevated G/S ratio. The Gas Chromatography-Mass Spectrometer analysis of low-molecular-weight compounds revealed that the delignification resulted in the formation of alcohols, organic acids, benzodiazepines, and alkanes. Identified benzodiazepines implied the degradation of G and S units of lignin. These findings will help to improve the efficiency of biodelignification and expand our understanding of its mechanism.