Lignin Subunits Research Articles

Optimum Nitrogen and Density Allocation for Trade−Off Between Yield and Lodging Resistance of Winter Wheat

Increasing nitrogen and planting density can enhance crop yield, but it can reduce lodging resistance due to decreased lignin content. There is an urgent need to find feasible measures to balance these conflicting factors. We conducted a two-year field experiment in Tai’an, Shandong Province, China, evaluated SN23 (lodging resistant) and SN16 (lodging sensitive), under three nitrogen applications (120 kg/ha, N1; 240 kg/ha, N2; 360 kg/ha, N3) and four planting densities (75 plants/m2, D1; 225 plants/m2, D2; 375 plants/m2, D3; 525 plants/m2, D4), with N2D2 as the control, and measured lodging resistance related indexes and yield. N2D3 (SN23) increased internode length by 0.40 cm, reduced fresh weight by 0.09 g, resulting in a bending moment reduction of 0.39 g/cm. Lignin, cellulose, and hemicellulose decreased by 18.27, 16.48, and 16.22 mg/g DW, while S and G lignin subunits decreased by 118.09 and 127.34 μg/g DW, and H subunit increased by 23.59 μg/g DW. Eventually, the breaking strength was reduced by 1.74 g/cm resulting in a reduction of 0.09 in the lodging resistance index. The yield reached 10.17 t/ha due to an increase in spike number by 100.33 plants/m2, achieving an optimal balance between yield and lodging resistance in this experiment. This study provides a viable solution for balancing lodging resistance and yield in winter wheat.

Whole genome analysis of 26 bacterial strains reveals aromatic and hydrocarbon degrading enzymes from diverse environmental soil samples

Polycyclic aromatic compounds and petroleum hydrocarbons (PHs) are hazardous pollutants and seriously threaten the environment and human health. However, native microbial communities can adapt to these toxic pollutants, utilize these compounds as a carbon source, and eventually evolve to degrade these toxic contaminants. With this in mind, we isolated 26 bacterial strains from various environmental soil samples. Utilizing whole genome shotgun sequencing and analyses of these genomes revealed that they all belong to a single phylum with seven genera and sixteen species, and displayed variable genome sizes with CDS features, % GC contents, and GC skews. The analysis of genome annotation predicted genes/enzymes related to aromatic compound degradation, including the metabolism of homogentisate, salicylate and gentisate catabolism, benzoate, biphenyl, and phenylpropanoid compound degradation, and protocatechuate branch of beta-ketoadipate pathways. The majority of enzymes were found to belong to species Achromobacter pulmonis A (16%) & Achromobacter mucicolens (15%), Pseudomonas citronellolis (10%), and Comamonas thiooxydans (8%). Conversely, the highest number of hydrocarbon-degrading enzymes were found to be annotated in the species Pseudomonas citronellolis (13%), Comamonas thiooxydans (9%), Acinetobacter variabilis (7%), Pseudomonas aeruginosa, and Pseudomonas E sp002113165 (6%). These enzymes were categorized as dioxygenase, monooxygenase, hydroxylase, dehydrogenase, hydrolase, decarboxylase, aldolase, etc., and were predicted to function for benzoate, benzene, toluene, naphthalene, xylene, phthalate & terephthalate, anthranilate, protocatechuate & homoprotocatechuate, salicylate, aerobic & anaerobic gallate, and lignin subunit degradation, and catechol meta & ortho-cleavage pathways. In the future, molecular and biochemical characterization of these enzymes, together with strain assays for their capacity to degrade various pollutants, will help to improve the bioremediation process for environmental contaminations.

Unraveling the dynamics of lignin chemistry on decomposition to understand its contribution to soil organic matter accumulation

Abstract Aims Plant inputs are the primary organic carbon source that transforms into soil organic matter (SOM) through microbial processing. One prevailing view is that lignin plays a major role in the accumulation of SOM. This study investigated lignin decomposition using wood from different genotypes of Populus tremula as the model substrate. The genotypes naturally varied in lignin content and composition, resulting in high and low lignin substrates. Methods The wood was inoculated with fresh soil and decomposition was interpreted through mass loss and CO2 produced during a 12-month lab incubation. Detailed information on the decomposition patterns of lignin was obtained by Two-dimensional Nuclear magnetic resonance (2D NMR) spectroscopy on four occasions during the incubations. Results The lignin content per se did not affect the overall decomposition and ~ 60% of the mass was lost in both substrates. In addition, no differences in oxidative enzyme activity could be observed, and the rate of lignin decomposition was similar to that of the carbohydrates. The 2D NMR analysis showed the oxidized syringyl present in the initial samples was the most resistant to degradation among lignin subunits as it followed the order p-hydroxybenzoates > syringyl > guaiacyl > oxidized syringyl. Furthermore, the degradability of β–O–4 linkages in the lignin varied depending on the subunit (syringyl or guaiacyl) it is attached to. Conclusions Our study demonstrates that lignin contains fractions that are easily degradable and can break down alongside carbohydrates. Thus, the initial differences in lignin content per se do not necessarily affect magnitude of SOM accumulation.

Production of hierarchical porous bio‑carbon based on deep eutectic solvent fractionated lignin nanoparticles for high-performance supercapacitor

Owing to the low synthetic cost, low toxicity for most types and outstanding lignin solubility, deep eutectic solvents (DESs) have been recently exploited in fractionation of lignocellulosic biomass. Unfortunately, the DES-fractionated lignin, as another major product, was usually ignored and disposed improperly. Accordingly, the present study employed DES (choline chloride:lactic acid) to fractionate natural biomass for producing highly dispersed lignin nanoparticles. The obtained nano-lignin was utilized to prepare electrode materials for supercapacitors for the first time, and a higher specific capacitance than those prepared from conventional lignin was acquired. The results demonstrated that the lignin fractionation rate could be promoted via extending treatment time at low temperatures, whereas the DES to biomass mass ratio might be the lignin dissolution-limiting factor at high temperature. Uniform lignin nanoclusters could be obtained after DES fractionation, and increasing fractionation temperature was beneficial for preparing nano-lignin in DES. The cleavages of lignin-carbohydrate complexes and lignin subunits might account for the formation mechanism of lignin oligomers. Meanwhile, the aggregation of monolignols could contribute to the growth of lignin nanoparticles. The one-step KOH activation could induce hierarchical pores in lignin fractionated from moderate residence time, whereas two-step method was advisable for more aggregated DES-lignin. Based on the directly activated DES-lignin with a high surface area of 3577.3 m2 g−1, the prepared supercapacitor exhibited a superior specific capacitance of 248.8 F g−1. This study aimed to provide a novel method for producing high-performance energy storage carbon materials based on DES fractionated lignin from lignocellulose.

Investigation of chemical linkages between lignin and carbohydrates in cultured poplar cambium tissues via double isotope labeling

Lignin precursor labeled with 13C (coniferin-13Cα), carbohydrate precursor labeled with D (6,6-D2-glucose) were put into cambium tissue stripped from a growing poplar. The tissue was further cultured in vitro for 18d. Then, the isotopic abundance was determined. The results showed that the labeled precursors could be normally involved in the formation of new xylem. The labeled new xylem tissue was fractionated by ionic liquid DMSO/TBAH system to obtain two components: glucan-lignin complex (GL) and xylan-lignin complex (XL). The X-ray diffraction (XRD) results indicated that the crystalline form of cellulose in the GL component was transformed from type I to type II after the ionic liquid separation. Then the GL and XL were purified and modified by enzymatic and chemical methods, and their structures were elucidated by nuclear magnetic resonance (NMR) spectroscopy. The results showed that lignin subunits in the cultured tissues were mainly connected by β-5 and β-O-4 linkages, of which the β-O-4 substructure unit predominated. Lignin and carbohydrates were mainly connected by acetal bonds, ether bonds, and ester bonds. Combined with the carbohydrate composition and XRD analysis results, the GL components also confirmed the existence of acetal bonds, ester bonds and ether bonds between lignin and cellulose.

Exploration of the chemical linkages between lignin and cellulose in poplar wood with 13C and Deuterium dual isotope tracer

Stable isotopes were used to label lignin and cellulose to accurately explain the chemical association between them in poplar wood. Both the 13C-labeled lignin precursors (coniferin-[Cα-13C] or coniferin-[Cγ-13C]) and the Deuterium-labeled cellulose precursor (UDP-glucose-[6,6-D2]) were added to a growing poplar plant. After 20 d cultivation, hemicellulose and lignin-carbohydrate complexes (LCC) were extracted from the newly-formed xylem by Björkman method, then the abundance of 13C and D were detected. It revealed that both lignin precursors and cellulose precursor could be efficiently deposited in the newly-formed xylem, while cellulose precursor was detected mainly deposited on cellulose. Then the LCC was degraded using cellulase and hemicellulase to obtain enzymatically degraded lignin-carbohydrate complexes (EDLCC). The 13C NMR，1H NMR and 2D NMR analyses of EDLCC clearly showed that lignin subunits in poplar wood were mainly connected through β-O-4, β-5, β-β structures, while cellulose was mainly connected to lignin by ether bonds, acetal bonds and ester bonds. The ether bonds and acetal bonds were between cellulose C6 and lignin Cα, while the ester bonds were between cellulose C6 and lignin Cγ. The study provides new evidence for the structural information between cellulose and lignin in poplar.

Heterologous Expression, Characterization, and Comparison of Laccases from the White Rot Causing Basidiomycete Cerrena Unicolor

Lignin is the most abundant renewable source of aromatics on earth, and conversion of it to chemicals and fuels is needed to build an economically viable renewable biofuels industry. Biological routes to converting lignin to fuels and chemicals involve depolymerizing lignin using lignin-degrading enzymes that catalyze the breaking of ether and carbon-carbon bonds in the phenolic and non-phenolic subunits of lignin. Laccases are a crucial class of lignin-degrading enzymes and are copper-containing enzymes capable of oxidizing electron-rich organic substrates such as lignin using molecular oxygen as an electron acceptor. The genome of <em>Cerrena unicolor</em> was recently added to the JGI MycoCosm database and has eight laccases. Two of these laccases, designated Lc1 and Lc2, predicted to have the highest likelihood for successful expression in soluble, active form were selected for characterization. Lc1 and Lc2, which share 65% sequence identity, were heterologously expressed in <em>Komagataella pastoris</em> (formerly <em>Pichia pastoris</em>), allowing characterization and comparison of their purified forms. Lc1 and Lc2 had half-lives of 16 min and 185 min at 60°C, respectively, and, based on molecular dynamics simulations, the longer half-life of Lc2 was due to an increased number and persistence of salt bridges compared to Lc1. Using model lignin-like dimers and a nanostructure-initiator mass spectrometry assay to quantify catalysis of specific bond-breaking events, both Lc1 and Lc2 had their highest activity at pH 3 and in combination with syringaldehyde as a mediator, with Lc1 having a higher catalytic efficiency of β-O-4' ether and C-C bond breaking. This comparative study demonstrates the diversity, including thermostability differences, of laccases from the same fungus, and improves our understanding of laccase catalyzed breaking of bonds commonly found in lignin, which will facilitate the developing this important class of enzymes for applications in the conversion of lignin to valuable bioproducts.

Determination of the Structures of Lignin Subunits and Nanoparticles in Solution by Small-Angle Neutron Scattering: Towards Improving Lignin Valorization.

Lignin nanoparticles (LNPs) are usually produced from lignin solution through supersaturation. The structure of the lignin in solution is still poorly understood due to structural variability of isolated lignins. Here, lignins were extracted from different plants to establish a general pattern of their structure in several lignin solvents. Lignin molecules (lignin subunits) and larger aggregates were observed in dimethyl sulfoxide (DMSO), ethylene glycol (EG) and 0.1 N NaOD solutions by small-angle neutron scattering (SANS). It was proposed that the aggregates were composed of lignin subunits with a higher molecular weight and a higher ratio of the aliphatic to phenolic hydroxyl groups. The size, shape, and compactness are important factors that affect the uses of the LNPs, which were obtained from the SANS data for the first time. A discrepancy in the radius between SANS and DLS was discovered, pointing to a large hydration shell around the LNPs in aqueous solutions. The cytotoxicity of the corncob lignin, kraft lignin, and their LNPs were measured and compared.

Biocatalytic valorization of lignin subunit: Screening a carboxylic acid reductase with high substrate preference to syringyl functional group

Lignin is inexpensive and the most abundant source of biological aromatics. It can be decomposed to three types of subunits, 4-hydroxybenzoic, vanillic and syringic acids, each of which can be valorized to value added compounds. Syringaldehyde is a versatile phenolic aldehyde implicated with multiple bioactive properties as well as intermediates for biofuels. Herein, fourteen microbial carboxylic acid reductases (CARs) were screened for the biocatalysis of the energetically unfavorable reduction of syringic acid to syringaldehyde. Nine CARs were positive to syringic acid reduction, among which Mycobacterium abscessus CAR exhibited the highest analytical yield of the product. By the optimization of the reaction condition, the whole-cell biocatalyst (i.e., recombinant Escherichia coli expressing the gene) successfully converted syringic acid to syringaldehyde with a yield of 90%. Furthermore, structural features of the screened CAR responsible for the specificity toward the syringyl subunit were analyzed that helps to further engineer the biocatalyst for improved performances.

Hydrothermal method-assisted synthesis of self-crosslinked all-lignin-based hydrogels

Lignin, as the most abundant aromatic biopolymer, is being widely studied to replace phenol and some other petroleum-based materials in the polymer industry. However, the low substitution of lignin and high levels of additives greatly limited the applications of lignin-based materials. Herein, we first propose a simple but effective hydrothermal method assisted synthesis for the fabrication of self-crosslinked lignin-based hydrogels (Lig-Scgel) with super-high-contents (75 wt%) of lignin and controllable mechanical properties. The self-crosslink mechanism was inspired by the repolymerization of lignins under a hydrothermal environment. The employment of self-condensation of lignin subunits in the synthesis of Lig-Scgel can significantly improve the degree of crosslinking, thereby greatly reducing the addition of toxic crosslinkers. The appearances, microstructures, crosslink densities, and mechanical properties of Lig-Scgels can be well controlled by simply altering the hydrothermal temperatures. This strategy not only promotes green and large-scale applications of lignin but also provides insights in the development of environment-friendly polymeric materials.

The maize low-lignin brown midrib3 mutant shows pleiotropic effects on photosynthetic and cell wall metabolisms in response to chilling

Maize (Zea mays L.) is one of the major cereal crops in the world and is highly sensitive to low temperature. Here, changes in photosynthetic and cell wall metabolisms were investigated during a long chilling exposure in inbred line F2 and a low-lignin near-isogenic brown midrib3 mutant (F2bm3), which has a mutation in the caffeic acid O-methyltransferase (COMT) gene. Results revealed that the plant biomass was reduced, and this was more pronounced in F2bm3. Photosynthesis was altered in both lines with distinct changes in photosynthetic pigment content between F2bm3 and F2, indicating an alternative photoprotection mechanism between lines under chilling. Starch remobilization was observed in F2bm3 while concentrations of sucrose, fructose and starch increased in F2, suggesting a reduced sugar partitioning in F2. The cell wall was altered upon chilling, resulting in changes in the composition of glucuronorabinoxylan and a reduced cellulose level in F2. Chilling shifted lignin subunit composition in F2bm3 mutant to a higher proportion of p-hydroxyphenyl (H) units, whereas it resulted in lignin with a higher proportion of syringyl (S) residues in F2. On average, the total cell wall ferulic acid (FA) content increased in both genotypes, with an increase in ether-linked FA in F2bm3, suggesting a greater degree of cross-linking to lignin. The reinforcement of the cell wall with lignin enriched in H-units and a higher concentration in cell-wall-bound FA observed in F2bm3 as a response to chilling, could be a strategy to protect the photosystems.

Cell Wall Composition Impacts Structural Characteristics of the Stems and Thereby the Biomass Yield.

Maize stalks support leaves and reproductive structures and functionally support water and nutrient transport; besides, their anatomical and biochemical characteristics have been described as a plant defense against stress, also impacting economically important applications. In this study, we evaluated agronomical and stem description traits in a subset of maize inbred lines that showed variability for cell wall composition in the internodes. Overall, a great proportion of lignin subunit G and a low concentration of p-coumaric acid and lignin subunit S are beneficial for greater rind puncture resistance and taller plants, with a greater biomass yield. Also, the greater the proportions of subunit H, the longer the internode. Finally, the lower the total hemicellulose content, the greater the rind puncture resistance. Our results confirmed the effect of the cell wall on agronomic and stalk traits, which would be useful in applied breeding programs focused on biomass yield improvement.

The Inducible Accumulation of Cell Wall-Bound p-Hydroxybenzoates Is Involved in the Regulation of Gravitropic Response of Poplar.

Lignin in Populus species is acylated with p-hydroxybenzoate. Monolignol p-hydroxybenzoyltransferase 1 (PHBMT1) mediates p-hydroxybenzoylation of sinapyl alcohol, eventually leading to the modification of syringyl lignin subunits. Angiosperm trees upon gravistimulation undergo the re-orientation of their growth along with the production of specialized secondary xylem, i.e., tension wood (TW), that generates tensile force to pull the inclined stem or leaning branch upward. Sporadic evidence suggests that angiosperm TW contains relatively a high percentage of syringyl lignin and lignin-bound p-hydroxybenzoate. However, whether such lignin modification plays a role in gravitropic response remains unclear. By imposing mechanical bending and/or gravitropic stimuli to the hybrid aspens in the wild type (WT), lignin p-hydroxybenzoate deficient, and p-hydroxybenzoate overproduction plants, we examined the responses of plants to gravitropic/mechanical stress and their cell wall composition changes. We revealed that mechanical bending or gravitropic stimulation not only induced the overproduction of crystalline cellulose fibers and increased the relative abundance of syringyl lignin, but also significantly induced the expression of PHBMT1 and the increased accumulation of p-hydroxybenzoates in TW. Furthermore, we found that although disturbing lignin-bound p-hydroxybenzoate accumulation in the PHBMT1 knockout and overexpression (OE) poplars did not affect the major chemical composition shifts of the cell walls in their TW as occurred in the WT plants, depletion of p-hydroxybenzoates intensified the gravitropic curving of the plantlets in response to gravistimulation, evident with the enhanced stem secant bending angle. By contrast, hyperaccumulation of p-hydroxybenzoates mitigated gravitropic response. These data suggest that PHBMT1-mediated lignin modification is involved in the regulation of poplar gravitropic response and, likely by compromising gravitropism and/or enhancing autotropism, negatively coordinates the action of TW cellulose fibers to control the poplar wood deformation and plant growth.

Response of Lignin Metabolism to Light Quality in Wheat Population.

The low red/far-red (R/FR) light proportion at the base of the high-density wheat population leads to poor stem quality and increases lodging risk. We used Shannong 23 and Shannong 16 as the test materials. By setting three-light quality treatments: normal light (CK), red light (RL), and far-red light (FRL), we irradiated the base internodes of the stem with RL and FRL for 7h. Our results showed that RL irradiation enhanced stem quality, as revealed by increased breaking strength, stem diameter, wall thickness and, dry weight per unit length, and the total amount of lignin and related gene expression increased, at the same time. The composition of lignin subunits was related to the lodging resistance of wheat. The proportion of S+G subunits and H subunits played a key role in wheat lodging resistance. RL could increase the content of S subunits and G subunits and the proportion of S+G subunits, reduce the proportion of H subunits. We described here, to the best of our knowledge, the systematic study of the mechanism involved in the regulation of stem breaking strength by light quality, particularly the effect of light quality on lignin biosynthesis and its relationship with lodging resistance in wheat.

Stacking AsFMT overexpression with BdPMT loss of function enhances monolignol ferulate production in Brachypodiumdistachyon.

SummaryTo what degree can the lignin subunits in a monocot be derived from monolignol ferulate (ML‐FA) conjugates? This simple question comes with a complex set of variables. Three potential requirements for optimizing ML‐FA production are as follows: (1) The presence of an active FERULOYL‐CoA MONOLIGNOL TRANSFERASE (FMT) enzyme throughout monolignol production; (2) Suppression or elimination of enzymatic pathways competing for monolignols and intermediates during lignin biosynthesis; and (3) Exclusion of alternative phenolic compounds that participate in lignification. A 16‐fold increase in lignin‐bound ML‐FA incorporation was observed by introducing an AsFMT gene into Brachypodium distachyon. On its own, knocking out the native p‐COUMAROYL‐CoA MONOLIGNOL TRANSFERASE (BdPMT) pathway that competes for monolignols and the p‐coumaroyl‐CoA intermediate did not change ML‐FA incorporation, nor did partial loss of CINNAMOYL‐CoA REDUCTASE1 (CCR1) function, which reduced metabolic flux to monolignols. However, stacking AsFMT into the Bdpmt‐1 mutant resulted in a 32‐fold increase in ML‐FA incorporation into lignin over the wild‐type level.

The effects of chemical and structural factors on the enzymatic saccharification of Eucalyptus sp. samples pre-treated by various technologies

Lignocellulosic biomass is an important renewable resource to produce biofuels, due to its abundance and high polysaccharide content. Bioconversion is generally achieved in two stages: (i) pre-treatment, to reduce biomass recalcitrance, and (ii) enzymatic cellulose hydrolysis into fermentable sugars. However, the key structural and chemical features of biomass which hinder enzymatic hydrolysis remains challenging to evaluate. In this study, we explored the effect of eight different pre-treatments on the enzymatic hydrolysis of a Eucalyptus sp. – a potential feedstock for biofuel production. The analytical data collected covered the mass fractions of cellulose, hemicellulose and lignin, the syringyl: guaiacyl (S/G) ratio of lignin, functional group determination, crystallinity index (CrI) and accessibility of cellulose. Enzymatic hydrolysis was then performed using an in-house optimised cellulolytic enzyme cocktail (either alone or supplemented with xylanase or laccase) to elucidate the key factors of biomass recalcitrance which influence its enzymatic hydrolysis. Enzymatic saccharification was improved by changes in Eucalyptus induced by steam explosion pre-treatment, such as substantial removal of hemicelluloses, increased cellulose accessibility, and disruption of cell wall architecture and exposure of fibres. Partial removal of lignin, increased S/G lignin subunit ratio and lowered cellulose crystallinity exhibited no significant positive effects on biomass enzymatic hydrolyzability. Thus, this study provides insight into important chemical and structural features related to biomass recalcitrance arising from various pre-treatment methods, which can ultimately be used as a platform for the development of more efficient conversion technologies for novel and competitive bio-refineries.

Elucidating compositional factors of maize cell walls contributing to stalk strength and lodging resistance

Lodging is one of the causes of maize (Zea mays L.) production losses worldwide and, at least, the resistance to stalk lodging has been positively correlated with stalk strength. In order to elucidate the putative relationship between cell wall, stalk strength and lodging resistance, twelve maize inbreds varying in rind penetration strength and lodging resistance were characterized for cell wall composition and structure. Stepwise multiple regression indicates that H lignin subunits confer a greater rind penetration strength. Besides, the predictive model for lodging showed that a high ferulic acid content increases the resistance to lodging, whereas those of diferulates decrease it. These outcomes highlight that the strength and lodging susceptibility of maize stems may be conditioned by structural features of cell wall rather than by the net amount of cellulose, hemicelluloses and lignin. The results presented here provide biotechnological targets in breeding programs aimed at improving lodging in maize.

Thermal decomposition of biomass wastes derived from palm oil production

The palm oil industry produces large amounts of biomass by-products, such as palm empty fruit bunch (EFB), mesocarp fiber (MF), and kernel shell (KS). The thermal behavior and decomposition of the oil palm biomass wastes have been studied by thermogravimetry/mass spectrometry (TG/MS) and pyrolysis–gas chromatography/mass spectrometry (Py-GC/MS). The determination of biopolymer and inorganic composition facilitated the interpretation of the thermal decomposition results. It was established that the chemical compositions of KS and MF were similar, while EFB contained significantly higher amount of cellulose and potassium, and decomposed in a narrower temperature range. In addition to the thermogravimetric curves (TG and DTG), the product distribution of pyrolyzates also reflected the compositional differences. Py-GC/MS and TG/MS experiments showed that the increasing potassium content of the samples reduced the intensity of 4-hydroxy-5,6-dihydro-2H-pyran-2-one, which is formed via depolymerization and dehydration of xylan units. Consequently, these experiments proved that beside cellulose, the depolymerization of hemicellulose was also hindered, while the dehydration, fragmentation, and charring reactions were catalyzed by potassium. Additionally, the evolution of the characteristic cellulose and lignin products shifted to lower temperatures. The most dominant aromatic pyrolysis product was phenol from each sample, which formed to a large extent from the abundant 4-hydroxybenzoate (4Hb) lignin subunits by scission and decarboxylation. KS also produced 4-hydroxybenzoic acid in significant amounts during pyrolysis at 450 °C confirming the presence of 4Hb subunits in lignin. The evolution of phenol started at 220 °C from the 4Hb moieties, and ended at about 600 °C from the residual phenolic units, while the release of guaiacol, vinylguaiacol, and syringol occurred between 300 and 450 °C as monitored by TG/MS. Some characteristic carbohydrate products were attributed to either hemicellulose or cellulose degradation based on the evolution profiles of KS and MF with separated carbohydrate decomposition steps. The comparison of the oil palm biomass wastes showed that EFB is supposed to be treated separately from KS and MF during the utilization process due to the different thermal behavior and composition.

Unraveling the role of maize (Zea mays L.) cell-wall phenylpropanoids in stem-borer resistance

The cell wall putatively plays a role in host-plant resistance to phytopathogens. Here, we investigated which cell wall-bound phenolic compounds have determining roles in maize (Zea mays) resistance to attack by the Mediterranean corn borer Sesamia nonagrioides (Lefèbvre). Diverse sets of maize genotypes having contrasting hydroxycinnamate contents and borer resistance levels were evaluated. The interdependent relationships among some cell wall-bound phenolic compounds, such as ferulic acid and its dimers, or p-coumaric acid and syringyl lignin subunits, were analyzed. Both p-coumaric acid and syringyl momoners showed significant negative correlations with damage, as assessed by tunnel lengths, caused by S. nonagrioides larvae. Thus, the use of cell wall-bound p-coumaric acid in pest-resistant crop breeding programs is advisable.

Structural and functional analysis of lignostilbene dioxygenases from Sphingobium sp. SYK-6

Lignostilbene-α,β-dioxygenases (LSDs) are iron-dependent oxygenases involved in the catabolism of lignin-derived stilbenes. Sphingobium sp. SYK-6 contains eight LSD homologs with undetermined physiological roles. To investigate which homologs are involved in the catabolism of dehydrodiconiferyl alcohol (DCA), derived from β-5 linked lignin subunits, we heterologously produced the enzymes and screened their activities in lysates. The seven soluble enzymes all cleaved lignostilbene, but only LSD2, LSD3, and LSD4 exhibited high specific activity for 3-(4-hydroxy-3-(4-hydroxy-3-methoxystyryl)-5-methoxyphenyl) acrylate (DCA-S) relative to lignostilbene. LSD4 catalyzed the cleavage of DCA-S to 5-formylferulate and vanillin and cleaved lignostilbene and DCA-S (∼106 M−1 s−1) with tenfold greater specificity than pterostilbene and resveratrol. X-ray crystal structures of native LSD4 and the catalytically inactive cobalt-substituted Co-LSD4 at 1.45 Å resolution revealed the same fold, metal ion coordination, and edge-to-edge dimeric structure as observed in related enzymes. Key catalytic residues, Phe-59, Tyr-101, and Lys-134, were also conserved. Structures of Co-LSD4·vanillin, Co-LSD4·lignostilbene, and Co-LSD4·DCA-S complexes revealed that Ser-283 forms a hydrogen bond with the hydroxyl group of the ferulyl portion of DCA-S. This residue is conserved in LSD2 and LSD4 but is alanine in LSD3. Substitution of Ser-283 with Ala minimally affected the specificity of LSD4 for either lignostilbene or DCA-S. By contrast, substitution with phenylalanine, as occurs in LSD5 and LSD6, reduced the specificity of the enzyme for both substrates by an order of magnitude. This study expands our understanding of an LSD critical to DCA catabolism as well as the physiological roles of other LSDs and their determinants of substrate specificity.