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Lignin, a major plant cell wall component, has an important role in plant-defense mechanisms against pathogens and is a promising renewable carbon source to produce bio-based chemicals. However, our understanding of microbial metabolism is incomplete regarding certain lignin-related compounds like p-coumaryl and sinapyl alcohols. Here, we reveal peripheral pathways for the catabolism of the three main lignin precursors (p-coumaryl, coniferyl, and sinapyl alcohols) in the plant pathogen Xanthomonas citri. Our study demonstrates all the necessary enzymatic steps for funneling these monolignols into the tricarboxylic acid cycle, concurrently uncovering aryl aldehyde reductases that likely protect the pathogen from aldehydes toxicity. It also shows that lignin-related aromatic compounds activate transcriptional responses related to chemotaxis and flagellar-dependent motility, which might play an important role during plant infection. Together our findings provide foundational knowledge to support biotechnological advances for both plant diseases treatments and conversion of lignin-derived compounds into bio-based chemicals.

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The first step in carbon (C) turnover, where senesced plant biomass is converted through various pathways into compounds that are released to the atmosphere or incorporated into the soil, is termed litter decomposition. This review is focused on recent advances of how solar radiation can affect this important process in terrestrial ecosystems. We explore the photochemical degradation of plant litter and its consequences for biotic decomposition and C cycling. The ubiquitous presence of lignin in plant tissues poses an important challenge for enzymatic litter decomposition due to its biological recalcitrance, creating a substantial bottleneck for decomposer organisms. The recognition that lignin is also photolabile and can be rapidly altered by natural doses of sunlight to increase access to cell wall carbohydrates and even bolster the activity of cell wall degrading enzymes highlights a novel role for lignin in modulating rates of litter decomposition. Lignin represents a key functional connector between photochemistry and biochemistry with important consequences for our understanding of how sunlight exposure may affect litter decomposition in a wide range of terrestrial ecosystems. A mechanistic understanding of how sunlight controls litter decomposition and C turnover can help inform management and other decisions related to mitigating human impact on the planet.

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The hydrothermal pretreatment process stands out as a pivotal step in breaking down the hemicellulosic fraction of lignocellulosic biomasses, such as sugarcane bagasse and eucalyptus sawdust. This pretreatment step is crucial for preparing these materials for subsequent processes, particularly in food applications. This technique aims to disintegrate plant wall components like cellulose, hemicellulose, and lignin, and facilitating access in later phases such as enzymatic hydrolysis, and ultimately making fermentable sugars available. In this study, sugarcane bagasse and eucalyptus sawdust biomass underwent hydrothermal pretreatment at specific conditions, yielding two key components: dry biomass and hemicellulose liquor. The primary focus was to assess the impact of hydrothermal pretreatment followed by enzymatic hydrolysis, using the Celic Ctec III enzyme cocktail, to obtain fermentable sugars. These sugars were then transformed into membranes via strain Gluconacetobacter xylinus bacterial biosynthesis. Notably, the addition of a nitrogen source significantly boosted production to 14.76 g/ in hydrolyzed sugarcane bagasse, underscoring its vital role in bacterial metabolism. Conversely, in hydrolyzed eucalyptus, nitrogen source inclusion unexpectedly decreased yield, highlighting the intricate interactions in fermentation media and the pivotal influence of nitrogen supplementation. Characterization of membranes obtained in synthetic and hydrolyzed media through techniques such as FEG-SEM, FTIR, and TGA, followed by mass balance assessment, gauged their viability on an industrial scale. This comprehensive study aimed not only to understand the effects of pretreatment and enzymatic hydrolysis but to also evaluate the applicability and sustainability of the process on a large scale, providing crucial insights into its feasibility and efficiency in practical food-related scenarios, utilizing nanocellulose bacterial (BNC) as a key component.

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Efforts are intensifying to identify new biofuel sources in response to the pressing need to mitigate environmental pollutants, such as greenhouse gases, which are key contributors to global warming and various worldwide calamities. Algae and microalgae present themselves as excellent alternatives for solid-gaseous fuel production, given their renewable nature and non-polluting characteristics. However, making biomass production from these organisms economically feasible remains a challenge. This article collates various studies on the use of lignocellulosic waste, transforming it from environmental waste to valuable organic supplements for algae and microalgae cultivation. The focus is on enhancing biomass production and the metabolites derived from these biomasses.

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A novel MADS-box transcription factor from Pinus radiata D. Don was characterized. PrMADS11 encodes a protein of 165 amino acids for a MADS-box transcription factor belonging to group II, related to the MIKC protein structure. PrMADS11 was differentially expressed in the stems of pine trees in response to 45° inclination at early times (1 h). Arabidopsis thaliana was stably transformed with a 35S::PrMADS11 construct in an effort to identify the putative targets of PrMADS11. A massive transcriptome analysis revealed 947 differentially expressed genes: 498 genes were up-regulated, and 449 genes were down-regulated due to the over-expression of PrMADS11. The gene ontology analysis highlighted a cell wall remodeling function among the differentially expressed genes, suggesting the active participation of cell wall modification required during the response to vertical stem loss. In addition, the phenylpropanoid pathway was also indicated as a PrMADS11 target, displaying a marked increment in the expression of the genes driven to the biosynthesis of monolignols. The EMSA assays confirmed that PrMADS11 interacts with CArG-box sequences. This TF modulates the gene expression of several molecular pathways, including other TFs, as well as the genes involved in cell wall remodeling. The increment in the lignin content and the genes involved in cell wall dynamics could be an indication of the key role of PrMADS11 in the response to trunk inclination.

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Turkey litter waste is lignocellulosic and keratinous, requiring prior enzymatic treatment to facilitate fiber hydrolysis and utilization by microorganisms in anaerobic digestion (AD) process. The understanding of the performance of microorganisms in AD can be facilitated through molecular biology and bioinformatics tools. This study aimed to determine the taxonomic profile and functional prediction of microbial communities in the AD of turkey litter waste subjected to enzymatic pretreatment and correlate it with operational parameters. The tests involved the use of turkey litter (T) at 25 g L-1 of volatile solids, a granular inoculum (S) (10% m/v), and the addition of cellulase (C), and pectinase (P) enzymes at four concentrations. The use of enzymes increased methane production by 19% (turkey litter, inoculum, and cellulase-TSC4) and 15% (turkey litter, inoculum, and enzymatic pectinase-TSP4) compared to the control (turkey litter and inoculum-TS), being more effective in TSC4 (667.52 mLCH4), where there was consumption of acetic, butyric, and propionic acids. The pectinase assay (TSP4) showed a methane production of 648 mLCH4 and there was the accumulation of metabolites. Cellulolytic microorganisms Bacteroides, Ruminofilibacter, Lachnospiraceae, Ruminococcaceae, and Methanosaeta were favored in TSC4. In TSP4, the predominant genus was Macellibacteroides and Methanosarcina, and genes involved in methylotrophic methanogenesis were also found (mtaB, mtmB, and mtbB). Enzymes involved in hydrogenotrophic methanogenesis were identified in both assays (TSC4 and TSP4). Molecular tools helped to understand the metabolic routes involved in AD with enzymatic treatment, allowing the elaboration of strategies to improve the sustainable degradation of turkey litter waste.

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The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.

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Cotreatment, mechanical disruption of lignocellulosic biomass during microbial fermentation, is a potential alternative to thermochemical pretreatment as a means of increasing the accessibility of lignocellulose to biological attack. Successful implementation of cotreatment requires microbes that can withstand milling, while solubilizing and utilizing carbohydrates from lignocellulose. In this context, cotreatment with thermophilic, lignocellulose-fermenting bacteria has been successfully evaluated for a number of lignocellulosic feedstocks. Here, cotreatment was applied to sugarcane bagasse using monocultures of the cellulose-fermenting Clostridium thermocellum and cocultures with the hemicellulose-fermenting Thermoanaerobacterium thermosaccharolyticum. This resulted in 76 % carbohydrate solubilization (a 1.8-fold increase over non-cotreated controls) on 10 g/L solids loading, having greater effect on the hemicellulose fraction. With cotreatment, fermentation by wild-type cultures at low substrate concentrations increased cumulative product formation by 45 % for the monoculture and 32 % for the coculture. These findings highlight the potential of cotreatment for enhancing deconstruction of sugarcane bagasse using thermophilic bacteria.

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Grapevine production is economically indispensable for the global wine industry. Currently, Mexico cultivates grapevines across approximately 28 500 hectares, ranking as the 26th largest producer worldwide. Given its significance, early detection of plant diseases' causal agents is crucial for preventing outbreaks. Consequently, our study aimed to identify fungal strains in grapevines exhibiting trunk disease symptoms and assess their enzymatic capabilities as indicators of their phytopathogenic potential. We collected plant cultivars, including Malbec, Shiraz, and Tempranillo, from Querétaro, Mexico. In the laboratory, we superficially removed the plant bark to prevent external contamination. Subsequently, the sample was superficially disinfected, and sawdust was generated from the symptomatic tissue. Cultivable fungal strains were isolated using aseptic techniques from the recovered sawdust. Colonies were grown on PDA and identified through a combination of microscopy and DNA-sequencing of the ITS and LSU nrDNA regions, coupled with a BLASTn search in the GenBank database. We evaluated the strains' qualitative ability to degrade cellulose, starch, and lignin using specific media and stains. Using culture morphology and DNA-sequencing, 13 species in seven genera were determined: Acremonium, Aspergillus, Cladosporium, Dydimella, Fusarium, Sarocladium, and Quambalaria. Some isolated strains were able to degrade cellulose or lignin, or starch. These results constitute the first report of these species community in the Americas. Using culture-dependent and DNA-sequencing tools allows the detection of fungal strains to continue monitoring for early prevention of the GTD.

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Laccases (EC 1.10.3.2) are oxidoreductases that belong to the multicopper oxidase subfamily and are classified as yellow/white or blue according to their absorption spectrum. Yellow laccases are more useful for industrial processes since they oxidize nonphenolic compounds in the absence of a redox mediator and stand out for being more stable and functional under extreme conditions. This study aimed to characterize a new laccase that was predicted to be present in the genome of Chitinophaga sp. CB10 - Lac_CB10. Lac_CB10, with a molecular mass of 100.06 kDa, was purified and characterized via biochemical assays using guaiacol as a substrate. The enzyme demonstrated extremophilic characteristics, exhibiting relative activity under alkaline conditions (CAPS buffer pH 10.5) and thermophilic conditions (80-90°C), as well as maintaining its activity above 50% for 5 h at 80°C and 90°C. Furthermore, Lac_CB10 presented a spectral profile typical of yellow laccases, exhibiting only one absorbance peak at 300 nm (at the T2/T3 site) and no peak at 600 nm (at the T1 site). When lignin was degraded using copper as an inducer, 52.27% of the material was degraded within 32 h. These results highlight the potential of this enzyme, which is a novel yellow laccase with thermophilic and alkaline activity and the ability to act on lignin. This enzyme could be a valuable addition to the biorefinery process. In addition, this approach has high potential for industrial application and in the bioremediation of contaminated environments since these processes often occur at extreme temperatures and pH values. IMPORTANCE: The characterization of the novel yellow laccase, Lac_CB10, derived from Chitinophaga sp. CB10, represents a significant advancement with broad implications. This enzyme displays exceptional stability and functionality under extreme conditions, operating effectively under both alkaline (pH 10.5) and thermophilic (80-90°C) environments. Its capability to maintain considerable activity over extended periods, even at high temperatures, showcases its potential for various industrial applications. Moreover, its distinctive ability to efficiently degrade lignin-demonstrated by a significant 52.27% degradation within 32 h-signifies a promising avenue for biorefinery processes. This newfound laccase's characteristics position it as a crucial asset in the realm of bioremediation, particularly in scenarios involving contamination at extreme pH and temperature levels. The study's findings highlight the enzyme's capacity to address challenges in industrial processes and environmental cleanup, signifying its vital role in advancing biotechnological solutions.

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Brazilian biomes are important sources for environmental microorganisms, including efficient metabolic machineries, like actinomycetes. These bacteria are known for their abilities to produce many bioactive compounds, including enzymes with multiple industrial applications. The present work aimed to evaluate lignocellulolytic abilities of actinomycetes isolated from soil and rhizosphere samples collected at Caatinga, Atlantic and Amazon Forest. Laccase (Lac), lignin peroxidase (LiP), manganese peroxidase (MnP) and cellulase were evaluated for their efficiency. These enzymes have an essential role in lignin decomposition, through oxidation of phenolic and non-phenolic compounds, as well as enzymatic hydrolysis of vegetal biomass. In this sense, a total of 173 actinomycetes were investigated. Eleven (11) of them were selected by their enzymatic performance. The actinomycete AC166 displayed some activity in all analysed scenarios in terms of Lac, MnP and LiP activity, while AC171 was selected as the most promising strain, showing the following activities: 29.7 U.L-1 for Lac; 2.5 U.L-1 for LiP and 23 U.L-1 for MnP. Cellulolytic activities were evaluated at two pH conditions, 4.8 and 7.4, obtaining the following results: 25 U.L-1 and 71 U.L-1, respectively. Thermostability (4, 30 and 60 o C) and salinity concentrations (0 to 4 M) and pH variation (2.0 to 9.0) stabilities of the obtained LiP and Lac enzymatic extracts were also verified. The actinomycete strain AC171 displayed an adaptable response in distinct pH and salt profiles, indicating that bacterial LiP was some halophilic type. Additionally, the strain AC149 produced an alkali and extreme halophilic lignin peroxidase, which are promising profiles for their future application under lignocellulosic biomass at bioethanol biorefineries.

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The agricultural sugarcane residues, bagasse and straws, can be used for second-generation ethanol (2GE) production by the cellulose conversion into glucose (saccharification). However, the lignin content negatively impacts the saccharification process. This polymer is mainly composed of guaiacyl (G), hydroxyphenyl (H), and syringyl (S) units, the latter formed in the ferulate 5-hydroxylase (F5H) branch of the lignin biosynthesis pathway. We have generated transgenic lines overexpressing ShF5H1 under the control of the C4H (cinnamate 4-hydroxylase) rice promoter, which led to a significant increase of up to 160% in the S/G ratio and 63% in the saccharification efficiency in leaves. Nevertheless, the content of lignin was unchanged in this organ. In culms, neither the S/G ratio nor sucrose accumulation was altered, suggesting that ShF5H1 overexpression would not affect first-generation ethanol production. Interestingly, the bagasse showed a significantly higher fiber content. Our results indicate that the tissue-specific manipulation of the biosynthetic branch leading to S unit formation is industrially advantageous and has established a foundation for further studies aiming at refining lignin modifications. Thus, the ShF5H1 overexpression in sugarcane emerges as an efficient strategy to improve 2GE production from straw.

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Two methods of sterilization of lignocellulosic biomass were performed in this study. Eucalypt waste (EW) supplemented with rice bran (RB) was added in the proportions 80:20 and 90:10 in dry weight. The compositions were sterilized by physical method (autoclaving) and by chemical method (H2O2). The production of extracellular enzymes by Lentinula edodes strains was compared within the two methods. Inactivation of catalase present in RB was achieved with 250 mM H2O2. The use of H2O2, when compared by physical method, favored high production of hydrolytic enzymes such as endoglucanase (1,600 IU/kg), twofold higher, ß-glucosidase (1,000 IU/kg), fivefold higher, xylanase (55,000 IU/kg), threefold higher and ß-xylosidase (225 IU/kg), similar results. Oxidative enzymes, MnP and laccase, were produced within a different profile between strains, with shorter times for laccase (2,200 IU/kg) by SJC in 45 days and MnP (2,000 IU/kg) by CCB-514 in 30 days. High production of extracellular enzymes is achieved by the use of the chemical method of sterilization of lignocellulosic biomass; in addition to no energy consumption, this process is carried out in a shorter execution time when compared to the physical process. The use of H2O2 in sterilization does not produce toxic compounds from the degradation of hemicellulose and cellulose such as furfural and hydroxy-methyl-furfural that cause inhibition of microorganisms and enzymes.

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Black wattle (Acacia mearnsii De Wild.) barks are residues produced by tannin industries in huge quantities, which are normally discharged on environmental or used for energy production. Therefore, this study aimed to evaluate the use of black wattle bark residues as a raw material on obtaining of a rich-cellulose material by alkaline (MET1), acetosolv (MET2), and organosolv (MET3) procedures. The results obtained indicated that the alkaline methodology, followed by a bleaching step (MET1), promoted klason lignin and hemicellulose removals more efficiently. It was possible to observe that better results were achieved using NaOH concentration of 6% (wt%), at 65 °C for 2.5 h, presenting a yield of 63.24 ± 1.25%, and a reduction on klason lignin content of almost 90.45%. Regarding the bleaching step, it was possible to obtain a material free of non-cellulosic compounds with a yield of 78.28 ± 1.48%. Thermogravimetric analysis indicated the removal of lignin and hemicellulose as well as an increase in cellulose degradation temperature, due to changes in crystalline phases. According to X-ray diffraction (XRD), the procedures employed have led to an increase in crystallinity from 66.27 to 91.78% due to the removal of non-cellulosic compounds. Scanning electron microscopy (SEM) showed morphological alterations in accordance with the removal of non-cellulosic compounds.

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Phenylpropanoids belong to a wide group of compounds commonly secreted by plants and involved in different roles related with plant growth and development and the defense against plant pathogens. Some key intermediates from shikimate pathway are used to synthesize these compounds. In this way, by the phenylpropanoid pathway several building blocks are achieved to obtain flavonoids, isoflavonoids, coumarins, monolignols, phenylpropenes, phenolic acids, stilbenes and stilbenoids, and lignin, suberin and sporopollenin for plant-microbe interactions, structural support and mechanical strength, organ pigmentation, UV protection and acting against pathogens. Some reviews have revised phenylpropanoid biosynthesis and regulation of the biosynthetic pathways. In this review, the most important chemical structures about phenylpropanoid derivatives are summarized grouping them in different sections according to their structure. We have put special attention on their different roles in plants especially in plant health, growth and development and plant-environment interactions. Their interaction with microorganisms is discussed including their role as antimicrobials. We summarize all new findings about new developed structures and their involvement in plants health.

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Marine-derived fungi have attracted much attention due to their ability to present a new biosynthetic diversity. About 50 fungal isolates were obtained from Tunisian Mediterranean seawater and then screened for the presence of lignin-peroxidase (LiP), manganese-dependent peroxidase (MnP), and laccase (Lac) activities. The results obtained from both qualitative and quantitative assays showed that four of marine fungi isolates had a high potential to produce lignin-degrading enzymes. They were characterized taxonomically by a molecular method, based on international spacer (ITS) rDNA sequence analysis, as Chaetomium jodhpurense (MH667651.1), Chaetomium maderasense (MH665977.1), Paraconiothyrium variabile (MH667653.1), and Phoma betae (MH667655.1) which have been reported as producers of ligninolytic enzyme in the literature. The enzymatic activities and culture conditions were optimized using a Fractional Factorial design (2 7- 4). Then, fungal strains were incubated with the addition of 1% of crude oil in 50% of seawater for 25 days to evaluate their abilities to simultaneously degrade hydrocarbon compounds and to produce ligninolytic enzymes. The strain P. variabile exhibited the highest crude oil degradation rate (48.3%). Significant production of ligninolytic enzymes was recorded during the degradation process, which reached 2730 U/L for the MnP, 410 U/L for LiP, and 168.5 U/L for Lac. The FTIR and GC-MS analysis confirmed that the isolates rapidly biodegrade crude oil under ecological and economic conditions.

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Global growth impacts on the increased use and demand for natural resources, requiring solutions for the high volume of industrial waste and by-products generated from the most diverse commercial areas, mainly the food sector. Among the main residues with a large volume generated, those from fruit processing, grain cleaning in processing units, vegetables, and discards from the animal production industry stood out. Approximately 1.3 billion all food produced worldwide is lost or wasted per year being fruits, vegetables, roots, and tubers responsible for about half of the total amount. Many of these by-products have interesting nutrients in their composition such as fibers, proteins, and bioactive compounds. An interesting example is the sugarcane bagasse. Fibrous residue, derived from sugarcane extraction, the bagasse represents about 30-34 % of the total sugarcane mass. This is one of the most abundant cellulosic residues and contains approximately 39 % of cellulose, 28 % of hemicellulose, and 18 % of lignin. Therefore, as well as the bagasse, several residues from agroindustrial can be considered promising alternative substrates, being valuable sources for the development of high-value-added products, such as biopolymers, bioenergy, and chemical products. In addition, the reuse of agroindustrial wastes may be considered an attractive option for reducing the environmental impact caused by their generation. In the case of biopolymers, the energy savings of bio-based polymers is around 20-50 GJ/t of polymer. In this review, we have selected two commercially promising approaches to the application and use of agroindustrial residues, aiming their use for biodegradable packaging and microbial polysaccharides bio-production, improving overall sustainability and economic aspects of the scientific research, technology and modern industry.

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A modification of the conventional batch organosolv process is proposed in a way where the solid biomass remains inside a basket, physically separated from the liquid phase, with the vapor promoting the fractionation of the biomass and the extracted compounds and fragments being washed down to the liquid phase. The modified organosolv process applied to sugarcane bagasse (SB-M) delivers a rich cellulosic solid phase that after enzymatic hydrolysis leads to a hydrolyzed with approximately 100 g L-1 of glucose. At the same enzymatic hydrolysis conditions, the conventional organosolv process (SB-C) delivers a hydrolyzed with 80 g L-1 of glucose, while the autohydrolysis process (SB-A) leads to 55 g L-1 of glucose. These different results are related to the cellulose content: SB-M (70%), SB-C (57%), e SB-A (44%), as well the reduced lignin content in the SB-M. The novelty of this study is the confirmation that it is possible to degrade lignin from sugarcane bagasse and simultaneously remove its fragments from the cellulose fibers in a batch reactor containing an internal basket. This study describes a simple and rapid protocol for the isolation of the main components of lignocellulosic biomass (cellulose, hemicellulose, and lignin), which may lead to the study of new catalysts for the chemical transformation of these components separately or simultaneously to the step of pretreatment.

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One of the critical steps of the biotechnological production of xylitol from lignocellulosic biomass is the deconstruction of the plant cell wall. This step is crucial to the bioprocess once the solubilization of xylose from hemicellulose is allowed, which can be easily converted to xylitol by pentose-assimilating yeasts in a microaerobic environment. However, lignocellulosic toxic compounds formed/released during plant cell wall pretreatment, such as aliphatic acids, furans, and phenolic compounds, inhibit xylitol production during fermentation, reducing the fermentative performance of yeasts and impairing the bioprocess productivity. Although the toxicity of lignocellulosic inhibitors is one of the biggest bottlenecks of the biotechnological production of xylitol, most of the studies focus on how much xylitol production is inhibited but not how and where cells are affected. Understanding this mechanism is important in order to develop strategies to overcome lignocellulosic inhibitor toxicity. In this mini-review, we addressed how these inhibitors affect both yeast physiology and metabolism and consequently xylose-to-xylitol bioconversion. In addition, this work also addresses about cellular adaptation, one of the most relevant strategies to overcome lignocellulosic inhibitors toxicity, once it allows the development of robust and tolerant strains, contributing to the improvement of the microbial performance against hemicellulosic hydrolysates toxicity. KEY POINTS: • Impact of lignocellulosic inhibitors on the xylitol production by yeasts • Physiological and metabolic alterations provoked by lignocellulosic inhibitors • Cell adaptation as an efficient strategy to improve yeast's robustness.

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