Lignin Degradation Products Research Articles

Antioxidant Properties of Pecan Shell Bioactive Components of Different Cultivars and Extraction Methods.

Pecan shells are a rich source of various bioactive compounds with potential antioxidant and antimicrobial properties. This study investigated the effect of pecan variety and method extraction on the antioxidant property of shell extracts. Twenty different varieties of pecan shells were subjected to either aqueous or ethanolic extraction and were examined for total phenolics and antiradical activity. The phenolic content and antiradical activity of shell extracts were significantly (p < 0.05) varied with different pecan cultivars. The total phenolic content of ethanol extracts ranged from 304.2 (Caddo) to 153.54 (Cherokee) mg GAE g−1 of dry extract and was significantly greater (p < 0.05) than those obtained by aqueous extraction. The antiradical activity of ethanol extracts ranged from 840.6 (Maramec) to 526.74 (Caper Fear) mg TE g−1, while aqueous extracts ranged from 934.9 (Curtis) to 468.3 (Elliot) mg TE g−1. Chemical profiling of the crude and acid hydrolyzed extracts was performed by reverse phase high performance liquid chromatography and flow injection electrospray ionization mass spectrometry. Lignin degradation products such as lignols, dilignols, trilignols, and oligolignols were found to be the major components of tested extracts. Phenolic content and antiradical activity of pecan shell extracts are significantly varied with cultivars and methods of extraction.

Characterization of ionized lignin model compounds with α-O-4 linkages by positive- and negative-ion mode electrospray ionization tandem mass spectrometry based on collision-activated dissociation.

The biggest obstacle in the rational conversion of biomass into aromatic chemicals is the identification of unknown compounds in lignin degradation mixtures that are highly complex. As opposed to lignin degradation products with β-O-4 linkages, very little is known about the mass spectrometric analysis of lignin degradation products with α-O-4 linkages. Lignin model compounds with an α-O-4 and another linkage, as well as lignin model compounds with only β-O-4 linkages, were ionized by attachment of lithium or sodium cations under positive-ion mode electrospray ionization (ESI) or by deprotonation in negative-ion mode ESI in a linear quadrupole ion trap mass spectrometer. The ions were subjected to collision-activated dissociation in multiple-stage tandem mass spectrometry experiments to characterize their fragmentation patterns. All studied compounds formed abundant sodium and lithium cation adducts in positive-ion mode ESI with no fragmentation. Model compounds with β-O-4 linkages displayed stable [M - H]- ions in negative-ion mode ESI whereas compounds with α-O-4 linkages only showed fragment ions. CAD of the lithiated α-O-4 compounds provided more structural information than CAD of sodiated compounds. However, both sodiated and lithiated compounds with α-O-4 linkages showed losses of monomer units at the MS2 stage, which is useful for sequencing of lignins with this type of linkage. An ionization and sequencing method has been developed for lignin model compounds with α-O-4 linkages that spontaneously fragment upon ionization via (-)ESI.

Utilization of the zebrafish model to unravel the harmful effects of biomass burning during Amazonian wildfires

Amazonian wildfires in 2019 have raised awareness about rainforest burning due to increased emissions of particulate matter and carbon. In the context of these emissions, by-products of lignin thermal degradation (i.e. methoxyphenols) are often neglected. Methoxyphenols entering the atmosphere may form intermediates with currently unknown reaction mechanisms and toxicity. This study for the first time provides a comprehensive insight into the impact of lignin degradation products [guaiacol, catechol], and their nitrated intermediates [4-nitrocatechol, 4,6-dinitroguaiacol, 5-nitroguaiacol] on zebrafish Danio rerio. Results revealed 4-nitrocatechol and catechol as the most toxic, followed by 4,6DNG > 5NG > GUA. The whole-organism bioassay integrated with molecular modeling emphasized the potential of methoxyphenols to inhibit tyrosinase, lipoxygenase, and carbonic anhydrase, consequently altering embryonic development (i.e. affected sensorial, skeletal, and physiological parameters, pigmentation formation failure, and non-hatching of larvae). The whole-organism bioassay integrated with in silico approach confirmed the harmful effects of lignin degradation products and their intermediates on aquatic organisms, emphasizing the need for their evaluation within ecotoxicity studies focused on aquatic compartments.

Lignin Degradation and Its Use in Signaling Development by the Coprophilous Ascomycete Podospora anserina.

The filamentous fungus Podospora anserina is a good model to study the breakdown of lignocellulose, owing to its ease of culture and genetical analysis. Here, we show that the fungus is able to use a wide range of lignocellulosic materials as food sources. Using color assays, spectroscopy and pyrolysis–gas chromatography mass spectrometry, we confirm that this ascomycete is able to degrade lignin, primarily by hydrolyzing β–O-4 linkages, which facilitates its nutrient uptake. We show that the limited weight loss that is promoted when attacking Miscanthus giganteus is due to a developmental blockage rather than an inefficiency of its enzymes. Finally, we show that lignin, and, more generally, phenolics, including degradation products of lignin, greatly stimulate the growth and fertility of the fungus in liquid cultures. Analyses of the CATΔΔΔΔΔ mutant lacking all its catalases, pro-oxidants and antioxidants indicate that improved growth and fertility of the fungus is likely caused by augmented reactive oxygen species levels triggered by the presence of phenolics.

Linking the unique molecular complexity of dissolved organic matter to flood period in the Yangtze River mainstream

Large rivers transport a significant amount of terrestrially derived dissolved organic matter (DOM) to coastal oceans, consisting of a critical component of the global biogeochemical cycle. Although high flow events usually introduce more terrestrial DOM than baseflow, the underlying molecular complexity and lability of DOM during high discharge are not well constrained, especially in large river ecosystems. By combining ultraviolet and fluorescent spectroscopy, and ultrahigh-resolution mass spectrometry, we found that stronger terrestrial DOM signal was detected during high discharge than normal discharge in the Yangtze River mainstream. The averaged DOC concentration was higher during high discharge than normal discharge. Optical properties confirmed higher aromaticity and relatively higher humic-like fluorescent components in DOM during high discharge. The molecular composition showed significantly higher molecular complexity, averaged molecular weight, aromaticity, relative abundances of polyphenols and highly unsaturated compounds of DOM during high discharge than normal discharge. A large set of unique molecular formulae (up to 4927) was only detected during high discharge. These unique molecular formulae were mostly lignin degradation products, likely due to more intensive soil leaching during high discharge. By comparing with incubation experiments and the Yangtze River mouth and East China Sea DOM molecular composition, some of these unique molecular formulae during high discharge are resistant to both bio- and photo-degradation, and persist during their transport to the East China Sea. Therefore, we suggest that high discharge will additionally introduce a relatively recalcitrant pool of DOM into the Yangtze River mainstream and persist during its journey to the ocean. Considering the projected increase of flood frequency, this study provides a preliminary foundation for further studies to better assess the underlying mechanisms how hydrology affect the biogeochemical cycling of DOM in large rivers.

Effects and Mechanisms of Alkali Recycling and Ozone Recycling on Enzymatic Conversion in Alkali Combined with Ozone Pretreatment of Corn Stover.

In order to minimize waste liquor, save resources, and reduce costs, the effects of alkali recycling and ozone recycling on enzymatic conversion in alkali combined with ozone pretreatment of corn stover and the mechanism were studied. The results showed that as the number of cycles of alkali/ozone filtrate increased, the enzymatic conversion and the loss of reducing sugars showed a downward trend. It was indicated that the ability of alkali to damage lignocellulosic decreased with an increasing number of alkali circulation and the accumulation of lignin degradation products generated during ozonolysis inhibited enzymatic conversion. When the ozone filtrate was recovered and used for hydrolysis directly, the enzymatic conversion rates were basically the same compared with the first self-circulation of ozone filtrate, and no sewage was discharged. In conclusion, the optimal circulating pretreatment was four times alkali circulation and ozone filtrate was used as an enzymolysis liquid directly, and the conversion rates of cellulose and hemicellulose were 85.96% and 34.26%, respectively, saving 44% alkali consumption at the same time. This paper provided the theoretical basis for the development of lignocellulose pretreatment technology with low cost, high efficiency, and high conversion rate.

Studies of the Fragmentation Mechanisms of Deprotonated Lignin Model Compounds in Tandem Mass Spectrometry.

Unlabeled and deuterium-labeled dimeric lignin model compounds with β-O-4 linkages were evaporated and ionized using negative ion mode electrospray ionization, transferred into a linear quadrupole ion trap, isolated, and subjected to collision-activated dissociation (CAD; MS2 experiments). The elemental compositions of the fragment ions were determined by using a high-resolution Orbitrap mass analyzer, and their structures were examined using further CAD experiments (MSn experiments wherein n = 2-5). Data analysis was facilitated by determining the fragmentation pathways for several deprotonated model compounds. The structures of the key fragment ions of several pathways were determined by comparison of the CAD mass spectra measured for undeuterated and deuterated analogues and for deprotonated authentic compounds. Some of the proposed reaction mechanisms were tested by examining additional deprotonated synthetic model compounds. Quantum chemical calculations were used to delineate the most likely reaction pathways and reaction mechanisms. This work provides basic information needed for the design of tandem mass spectrometry-based CAD sequencing strategies for mixtures of lignin degradation products.

Elimination of Cr(VI) from chromium slag with poplar lignin by electrochemical treatment in sulfuric acid solution.

In this paper, we proposed a novel method to eliminate nocuous Cr(VI) from chromium slag with poplar lignin by electrochemical treatment in sulfuric acid solution. In this electrochemical process, self-made Ti/SnO2-Sb anode and graphite cathode were applied, and the oxidative degradation of lignin proceeded simultaneously with the reduction of Cr(VI) in one pot. The influences of pivotal factors on electrocatalytic redox efficiency were investigated, such as chromium slag concentration, lignin concentration, current density, sulfuric acid concentration, and reaction time. The results showed that the elimination rate of Cr(VI) in chromium slag was 97.16± 1.13% and the total yield of lignin degradation products reached 93.78g/kg lignin under the optimal conditions. X-ray photoelectron spectroscopy (XPS), energy dispersive X-ray spectroscopy (EDS), and UV-visible spectrophotometer studies confirmed that most of the Cr(VI) ions were reduced to Cr(III) ions with the aid of lignin, and a small amount of Cr(VI) ions were adsorbed by lignin residue. Importantly, this method provides a typical example of "waste control by waste", which is treating waste chromium slag with waste lignin that can be an effective way to eliminate Cr(VI).

Understanding the formation of phenolic monomers during fractionation of birch wood under supercritical ethanol over iron based catalysts

The liquefaction of biomass in ethanol, at the critical point, has high potential due to low temperature and pressure (243 °C, 63 bar) when compared with water (374 °C, 220 bar). The current study deals with the fractionation of birch wood powder which was liquefied under supercritical ethanol over acidic or non-acidic catalysts, 5 wt % Fe-Beta-H-150 and 5 wt % Fe–SiO2, respectively. Based on the results, the reaction mechanism for the formation of lignin degradation products was proposed. The main phenolic product was isoeugenol over 5 wt % Fe-Beta-H-150 while intermediate products, i.e. such as coniferyl, and sinapyl alcohol, 4-propenyl syringol, syringaresinol, as well as syringyldehyde reacted rapidly further. The thermodynamic analysis was performed by Joback approach and using Gibbs-Helmholtz equation supporting the obtained results.

Bioethanol Production from Alkali Steam Explosion of Oil Palm of Empty Fruit Bunch Fiber

Bioethanol is the most promising biofuels and can be generated from lignocellulosic biomasses. The study of bioethanol production from oil palm empty fruit bunch (OPEFB) pretreated by alkali steam explosion was carried out employing separate enzymatic hydrolysis (SHF) and Simultaneous Saccharification and Fermentation (SSF) approaches. Alkaline extraction after steam explosion pretreatment of OPEFB led to the partial removal of lignin, hemicellulose and other degradation products from fiber surface. The pretreatment process is of great importance to ethanol yield. In the present study, the pretreatment was processed using a steam explosion reactor at 150°C, 4 bars for 30 minute with alkaline concentrations of NaOH 10% (kg/L). After pretreatment, the OPEFB substrates 10% and 15% (w/v) was saccharification using 30 FPU of Ctec2 and Htec2 enzyme and fermentation using yeast of Saccharomyces cerevisiae for 72 hours. The glucose obtained after 72 hours was 137.80 g/L (subtrate 15%) and 97.90 g/L (subtrate 10%). Bioethanol production in SSF mode faster and slightly more efficient process than SHF.

Gas-phase hydrodeoxygenation of phenol over Zn/SiO2 catalysts: Effects of zinc load, temperature, weight hourly space velocity, and H2 volumetric flow rate

The hydrodeoxygenation (HDO) of phenol catalyzed by Zn/SiO2 under atmospheric H2 pressure was investigated in a continuous fixed bed reactor. The effects of several process parameters (zinc load, reaction temperature, weight hourly space velocity (WHSV), and H2 volumetric flow rate) were evaluated to optimize process conditions. Phenol was selected as a stable model component for lignin degradation products in fast pyrolysis bio-oil. Silica-supported zinc catalysts were prepared with different loadings of the active metal (0.5%, 1%, 2%, 3%, and 4%) and assessed using characterization techniques such as XRD, ICP-OES, BET, H2-TPR/TPD, and FESEM–EDX. Reaction products including benzene, cyclohexene, and cyclohexane were identified through GC/FID analysis. Experimental results revealed that process yield increased with reaction temperature, metal loading, and WHSV. The selectivity percentages of the products were slightly changed by varying process parameters. Moreover, H2 volumetric flow rate exerted a negligible effect on product yield and selectivity.

Dearomatization-Rearomatization Strategy for Synthesizing Carbazoles with 2,2'-Biphenols and Ammonia by Dual C(Ar)-OH Bond Cleavages.

Carbazole is an essential building block in various pharmaceuticals, agrochemicals, natural products, and materials. For future sustainability, it is highly desirable to synthesize carbazole derivatives directly from renewable resources or cheap raw materials. Phenolic compounds are a class of degradation products of lignin. On the other hand, ammonia is a very cheap industrial inorganic chemical. Herein, an efficient dearomatization-rearomatization strategy has been developed to directly cross-couple 2,2'-biphenols with ammonia by dual C(Ar)-OH bond cleavages. This strategy provides a greener pathway to synthesize valuable carbazole derivatives from phenols.

Direct Inlet Probe Atmospheric Pressure Photo and Chemical Ionization Coupled to Ultrahigh Resolution Mass Spectrometry for the Description of Lignocellulosic Biomass.

Lignocellulosic biomass, in particular wood, is a complex mixture containing cellulose, hemicellulose, lignin, and other trace compounds. Chemical analysis of these biomasses, especially lignin components, is a challenge. Lignin is a highly reticulated polymer that is poorly soluble and usually requires chemical, enzymatic, or thermal degradation for its analysis. Here, we studied the thermal degradation of lignocellulosic biomass using a direct insertion probe (DIP). The DIP was used with two ionization sources: atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) coupled to ultrahigh-resolution mass spectrometry. Beech lignocellulosic biomass samples were used to develop the DIP-APCI/APPI methodology. Two other wood species (maple and oak) were analyzed after optimization of DIP parameters. The two ionization sources were compared at first and showed different responses toward beech samples, according to the source specificity. APPI was more specific to lignin degradation compounds, whereas APCI covered a larger variety of oxygenated compounds, e.g., fatty acids and polyphenolics compounds, in addition to lignin degradation products. The study of the thermodesorption profile gave information on the different steps of lignocellulosic biomass pyrolysis. The comparison of the three feed sample types (oak, maple, and beech), using principal component analysis (PCA) with DIP-APCI experiments, showed molecular level differences between beech wood pellets and the two other wood species (maple and oak).

Functional genomic analysis of bacterial lignin degraders: diversity in mechanisms of lignin oxidation and metabolism.

Although several bacterial lignin-oxidising enzymes have been discovered in recent years, it is not yet clear whether different lignin-degrading bacteria use similar mechanisms for lignin oxidation and degradation of lignin fragments. Genome sequences of 13 bacterial lignin-oxidising bacteria, including new genome sequences for Microbacterium phyllosphaerae and Agrobacterium sp., were analysed for the presence of lignin-oxidising enzymes and aromatic degradation gene clusters that could be used to metabolise the products of lignin degradation. Ten bacterial genomes contain DyP-type peroxidases, and ten bacterial strains contain putative multi-copper oxidases (MCOs), both known to have activity for lignin oxidation. Only one strain lacks both MCOs and DyP-type peroxidase genes. Eleven bacterial genomes contain aromatic degradation gene clusters, of which ten contain the central β-ketoadipate pathway, with variable numbers and types of degradation clusters for other aromatic substrates. Hence, there appear to be diverse metabolic strategies used for lignin oxidation in bacteria, while the β-ketoadipate pathway appears to be the most common route for aromatic metabolism in lignin-degrading bacteria.

Off-gassing from pilot-scale torrefied pine chips: Impact of torrefaction severity, cooling technology, and storage times

During handling and storage of conventional wood pellets, O2 depletion as well as CO and CO2 off-gassing can reach acutely hazardous levels and certain Volatile Organic Compounds (VOCs) may reach concerning levels from an occupational health and safety perspective. With new thermally pre-treated biomass commodities entering consumer markets, corresponding knowledge is needed on these assortments' off-gassing behaviour. In this study, relative concentrations of VOCs, CO, CO2, and O2 in the closed storage space of five different pilot-scale torrefied pine wood chip assortments were monitored over 12 days. The VOCs composition in the storage space differed between torrefaction treatment settings; terpenes decreased while furans and lignin degradation products peaked at narrow ranges with increased torrefaction severity, indicating that VOC off-gassing composition of individual compounds is highly specific. Generally, VOC amounts decreased with storage time, but for the mildest torrefied chips certain VOCs increased, predominantly compounds of higher volatility such as hexanal, acetone, and 2-pentylfuran. Also, the newly produced torrefied chips were cooled with two different post-process technologies: i) heat exchanging, and ii) heat exchanging with additional water spraying. Water spraying resulted in higher VOC concentrations, stronger O2 depletion, and factor four higher concentration of CO2 in the storage headspace.

The effect of lignin degradation products on the generation of pseudo-lignin during dilute acid pretreatment

Pseudo-lignin is an insoluble material that tends to deposit on acidic pretreated fiber surface and negatively impacts the biological conversion of biomass to valued added products. Employing a laboratory generated acidic hydrolysate solution from dilute acid pretreatment of bamboo residues, we sought to seek an improved understanding of pseudo-lignin formation from fragmented carbohydrates. In addition, we also performed experiments to investigate how lignin model compounds interact with carbohydrates en route to pseudo-lignin’s formation. The yields of pseudo-lignin generated from xylose (CXPL) were higher than those generated from glucose (CGPL). Gas chromatography-Mass spectrometry and Foloin-Ciocalteau analysis suggested that there were more aromatic compounds in hydrolyzate during CGPL’s formation. The presence of lignin phenolic model compounds had both a positive and negative effect upon quantitative yields of pseudo-lignin. Gel permeation chromatography and NMR analysis (31P and 2D-HSQC NMR) of the pseudo-lignin samples generated in the presence and absence of 4-hydroxybenzoic acid (4-HA), employed as a lignin model compound, revealed drastic differences in molecular weight, changes in hydroxyl group content, and hydrocarbon bonds, suggesting significant differences in the pseudo-lignin reaction pathways when certain lignin model compounds are present. Our findings demonstrate the effects of both hemicellulose carbohydrate makeup and lignin model compound impact on the formation of pseudo-lignin and its chemical properties, suggesting a potential way to minimize pseudo-lignin formation.

Assessment of molecular diversity of lignin products by various ionization techniques and high-resolution mass spectrometry

Lignin is a highly complex, plant-derived natural biomass component, the analysis of which requires significant demands on the analytical platform. Fourier transform ion cyclotron mass spectrometry (FT-ICR MS) has been shown to be able to readily assess the complexity of lignin and lignin degradation products by assigning tens of thousands of compounds with elemental formulae. Nevertheless, many experimental and instrumental parameters introduce discrimination towards certain components, which limits the comprehensive MS analysis. As a result, a complete characterization of the lignome remains a challenge. The present study investigated a degraded lignin sample using FT-ICR MS and compared several atmospheric pressure ionization methods, e.g., electrospray ionization, atmospheric-pressure chemical ionization, and atmospheric-pressure photoionization. The results clearly show that the number of heteroatoms (e.g., N, S, P) in the sample greatly increases the chemical diversity of lignin, while at the same time also providing potentially useful biomarkers. We demonstrate here that FT-ICR MS was able to directly isolate isotopically pure single components from the ultra-complex mixture for subsequent structural analysis, without the time-consuming chromatographic separation. CapsuleVarious ionization techniques coupled to FT-ICR MS provide a powerful tool to assess the lignome coverage.

Detection and Identification of Lignosulfonate Depolymerization Products Using UPLC-QTOF-MS and a Self-Built Database

In this study, based on depolymerization solution of commercially available sodium lignosulfonate under mild conditions, an efficient method for the chromatographic separation of lignosulfonate depolymerization products was firstly established. The self-built database contains 18 lignin-based standards and the information on their names, formulas, structures, retention times, fragments, and corresponding MSE data was developed. By searching the online spectral data in a novel informatics database, a total of nine aromatic lignin depolymerization products were identified automatically. Thus, a fast standard protocol for detection and identification of real lignin degradation products was preliminary built.

H3PW12O40/ZrO2 and 1-Butyl-3-methylimidazolium Chloride: A Double-Effect Catalyst for the Degradation of Alkali Lignin.

Alkali lignin is a component of the waste black liquor produced by the paper-making industry that is difficult to degrade. In recent years, the biological activities of lignin, such as free-radical scavenging and antioxidant capacity, have received increasing attention. Here, we prepared H3PW12O40/ZrO2 and used this catalyst together with the ionic liquid 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) as a double-effect catalyst for the degradation of alkali lignin. Single-factor and orthogonal tests showed that the best degradation conditions were as follows: reaction time, 2 h; reaction temperature, 100 °C; mass ratio of H3PW12O40/ZrO2 to lignin, 1:4; and substrate concentration, 2.5%. The phenolic hydroxyl group content of the lignin degradation product increased by 231.2% and the total hydroxyl group content increased by 337.1% when the double-effect catalyst was used rather than [BMIM]Cl alone. Analysis by gel permeation chromatography showed that both the weight-average molecular weight and the number-average molecular weight of the product were reduced and that the lignin was degraded into small-molecular-weight compounds by the macromolecule. The product after the catalytic degradation of lignin showed a markedly increased antioxidant capacity, which was similar to that of the commercial antioxidant, 2,6-ditert-butyl-4-methylphenol. The study opens up a new direction for the better utilization of lignin.

Application of Membrane Separation for Cleaning and Concentration of Nanolignin Suspensions in a Biorefinery Environment

Biorefining is a key technology for the sustainable production of chemicals, fuels and energy from abundant lignocellulosic resources. Within the concept, production of micro- and nanoscaled lignin particles recently gained increasing interest due to their excellent properties and potential application in high-value products. Fabricated via direct precipitation of Organosolv extracts, Nanolignin suspensions need to be increased in concentration and cleaned from impurities. Additionally, solvent background needs to be exchanged from hydroalcoholic solution to pure water for most applications. This work focuses on the application of ultrafiltration in diafiltration mode for this separation task. Experimental operation on lab-scale apparatuses demonstrate the reduction of Ethanol and impurities like sugars, dissolved lignin and sugar degradation products accessed by TOC analysis. Most important, the size of Nanolignin particles is not changed during 9 h of operation. Nevertheless, significant performance degradation due to compaction of polymeric membrane material as well as fouling resulting from deposited lignin particles has been monitored. Consequently, a rigorous optimisation of operating pressure and the development of an adequate membrane regeneration procedure is mandatory.