Search Results (99)

Lentinula edodes (Berk.) Pegler, also known as Shiitake, is one of the most popular edible mushroom species containing high contents of polysaccharides, proteins and unique aroma, widely cultivated in China, Japan and Korea. A series of studies has been carried out on the extraction and active effect of the L. edodes polysaccharides, but the molecular mechanisms involved in the protein expression profiles during the whole life cycle are relatively unclear. This study employed an iTRAQ-MS/MS proteomic approach, combined with real-time quantitative PCR (qRT-PCR) and enzyme activity assays, to systematically analyze the protein expression profiles and their relationship with lignocellulose degradation in L. edodes across four key developmental stages: mycelia (SF), brown film formation (BF), primordia (YF), and fruiting bodies (MF). A total of 2043 proteins were identified, with 1188 being differentially expressed proteins (DEPs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that metabolic processes, carbohydrate metabolism, and related pathways were significantly active during development. The study specifically focused on carbohydrate-active enzymes (CAZymes), identifying 197 CAZyme proteins classified into 78 families. Key families such as glycoside hydrolases (GHs) and carbohydrate esterases (CEs) played crucial roles in lignocellulose degradation. The enzymatic activities of major lignin-degrading enzymes (laccase, manganese peroxidase, and lignin peroxidase) were dynamically regulated across the developmental stages. qRT-PCR results largely corroborated the proteomic data, confirming the reliability of the protein expression profiles. This study provides a comprehensive, stage-resolved proteomic landscape of lignocellulose degradation during L. edodes development, revealing species-specific temporal dynamics, offering a valuable basis for understanding its growth and development, with implications for edible fungus cultivation and biomass conversion applications. Full article

Brewer’s spent grain (BSG) is an abundant lignocellulosic by-product whose valorization can support circular bioeconomy strategies. This study evaluated BSG bioconversion by Aspergillus oryzae ATCC 10124 under solid-state fermentation (SSF) to produce lignocellulolytic enzymes and release second-generation (2G) sugars relevant to biorefinery applications. SSF was monitored over 0–10 days, and FPase, endo-cellulase, β-glucosidase, xylanase, mannanase, amylase, and ligninolytic enzyme activities were quantified. Enzymatic crude extracts were further assessed in SDS-PAGE analysis. Glucose, cellobiose, xylose and arabinose release and consumption were tracked throughout fermentation, and substrate transformation was supported by FTIR. The secretome exhibited a predominantly hydrolytic profile, with maximal hemicellulolytic and cellulolytic activity around days 2–4, as well as sustained amylase activity. Ligninolytic activity was not detected. Sugar profiles indicated rapid early hydrolysis of glucose, followed by progressive pentose release. The stabilization and decline were consistent with fungal uptake. Changes in the carbohydrate fingerprint and SDS–PAGE banding supported structural polysaccharide remodeling and hydrolytic protein secretion. Thus, this SSF platform confirmed certain potential for low-cost cellulolytic and hemicellulolytic enzyme generation. However, because sugar accumulation was temporary and followed by consumption, this system is best interpreted as a biological pretreatment and enzyme-generation step that supports subsequent downstream valorization. Full article

We report on the isolation and comprehensive genomic and biochemical characterization of Aspergillus fumigatus VP2T, a thermophilic filamentous fungus recovered from Himalayan Forest soil with exceptional lignocellulolytic capacity. Whole-genome sequencing revealed a 32.1 Mb genome encoding 12,675 predicted genes, including an extensive repertoire of >300 carbohydrate-active enzymes (CAZymes). Notably, the genome harbors multiple auxiliary activity enzymes, including AA9-family lytic polysaccharide monooxygenases and several cellobiose dehydrogenases (CDHs), supporting oxidative–hydrolytic synergism during biomass degradation. Submerged fermentation using a cellulose–wheat bran–rice straw substrate induced high enzyme titers, including 33 U/mL endoglucanase and 131 U/mL CDH, exceeding activities commonly reported for both native and engineered fungal strains. Although exoglucanase (0.02 U/mL) and xylanase (14.22 U/mL) activities were comparatively modest, the strain VP2T demonstrated superior hydrolysis of untreated rice straw, achieving a 1.89-fold increase in saccharification efficiency relative to the commercial enzyme cocktail Cellic^® CTec2. Scanning electron microscopy confirmed extensive disruption of lignocellulosic architecture, consistent with enhanced enzyme accessibility and oxidative fiber loosening. Collectively, genomic evidence and functional assays identify A. fumigatus VP2T as a redox-optimized, moderately thermophilic biocatalyst suited for low-pH lignocellulose conversion. This study highlights the value of exploring thermophilic fungal biodiversity to discover native strains with inherent oxidative capacity, offering promising alternatives to pretreatment-intensive biorefinery processes and informing the rational development of tailored enzyme systems. Full article

The physiological efficacy of plant-based matrices is often limited because bioactive compounds are sequestered within complex lignocellulosic architectures, restricting their release and downstream activity. Fermentation-driven enzymatic biotransformation can overcome these structural barriers; however, the mechanisms by which fermentation-derived, non-viable functional ingredients (postbiotics) confer benefits remain incompletely defined. Here, we examined whether a postbiotic-rich, co-fermented plant matrix enhances host resilience under metabolic stress and whether such effects are accompanied by a remodeling of gut microbial functional capacity. A functional plant matrix was produced by solid-state co-fermentation using two Lactobacillus plantarum strains selected for complementary lignocellulolytic profiles. Untargeted metabolomics and deep shotgun metagenomic sequencing were integrated with a hydrocortisone-induced murine metabolic stress model to quantify substrate remodeling, host neuroendocrine/behavioral outcomes, and microbiome functional reprogramming. Co-fermentation markedly remodeled the phytochemical landscape, increasing extractable flavonoids and generating distinct metabolite clusters. In vivo, administration of the postbiotic-rich matrix partially normalized stress-responsive neuroendocrine markers (ACTH, TRH, and testosterone) and improved behavioral outcomes in open-field and forced swim assays. These systemic changes were paralleled by a coordinated shift in microbial functional potential, including the enrichment of carbohydrate-active enzyme (CAZyme) families involved in complex polysaccharide utilization (e.g., AA9, GH129, CE14) and attenuation of phosphotransferase system modules and cytochrome P450-related functions. Enzymatic synergy-driven biotransformation yields a postbiotic-rich functional matrix that is associated with a selective remodeling of gut microbiome metabolic potential under stress and concomitant improvement in host physiological resilience. This study underscores microbial functional remodeling as a critical mechanistic interface linking fermentation-modified substrates to host physiological recovery, providing a molecular framework for the development of targeted postbiotic interventions. Full article

This study evaluated the potential of sawdust as an inducer of carbon for the production of lignocellulolytic (cellulase and xylanase) enzymes by filamentous fungi for biofuel applications. Fourteen soil samples were collected from a phosphate mine in Phalaborwa, South Africa. Filamentous fungi were isolated from these samples using cellulose and xylose media. These isolateswere tested qualitatively and quantitatively for endocellulase and xylanases. Four isolates were found to have promising xylanase activity and these were identified as Amesia atrobrunnea, Penicillium citrinum, Rhizopus azygosporus and Aspergillus quadrilineatus using ITS1/2 sequencing. A time-course assay for xylanase activity revealed that R. azygosporus and A. quadrilineatus exhibited the highest activity. The crude enzymes were extracted from these strains and used for the enzymatic saccharification of the untreated sawdust. The total reducing sugars were estimated using the DNS method. The results for the enzymatic saccharification showed that a high total reducing sugar concentration of 2.35 g/L was released by 20 U/g of crude xylanases from A. quadrilineatus after 60 h of hydrolysis, while the synergistic hydrolysis of sawdust with the commercial cellulase, Celic CTec2, and the crude enzyme of 6 U/g from A. quadrilineatus showed the maximum concentration of total reducing sugars of 1.41 g/L after 72 h. Sawdust proved to be an effective inducer of xylanase production, although it was less effective for cellulase. This study reports that the commercial cellulase was outperformed by the crude enzymes during hydrolysis, highlighting the possibility that commercial enzymes may be replaced by microbial enzymes, thus lowering the environmental hazards of chemical formulations. Sugar yields could potentially be improved through biomass pretreatment, enzyme purification and strain improvement. Full article

Agave bagasse is a lignocellulosic residue generated after the extraction of fermentable sugars from agave hearts during tequila production. More than 0.5 million tons are generated annually, accumulating on a massive scale and posing a serious environmental challenge. In this regard, the objective of this study was to evaluate the degradative capacity of Trametes versicolor (Tv), Trametes hirsuta (Th), Irpex lacteus (Il), and Schizophyllum commune (Sc) on Agave tequilana Weber variety azul bagasse through the analysis of total sugars, cellulose, hemicellulose, and lignin reduction in a solid-static treatment. Under sterile conditions, Tv reduced total sugars by 95.0%, Th by 89.5%, Il by 91.8%, and Sc by 74.6%; whereas under non-sterile conditions, reductions were 81.6%, 71.4%, 84.9%, and 64.7%, respectively. Regarding structural fractions under sterile conditions, Tv showed reductions of 67.8% in cellulose, 61.9% in hemicellulose, and 68.8% in lignin. Th achieved 62.8%, 58.8%, and 66.1%, respectively; Il exhibited the highest values, with 72.9%, 66.9%, and 74.6%; while Sc recorded 55.9%, 44.2%, and 61.0%. In contrast, reductions were lower under non-sterile conditions: Tv recorded 57.8%, 34.2%, and 62.2%; Th, 53.9%, 32.1%, and 59.6%; Il, 58.8%, 47.1%, and 64.7%; and Sc, 49.9%, 30.0%, and 56.5%. Overall, sterile substrate conditions maximized lignocellulosic degradation; however, the sustained activity observed under non-sterile conditions demonstrates that effective biological pretreatment can be achieved without sterilization, which is more relevant for large-scale solid-state fermentation. The results demonstrate that T. versicolor and I. lacteus possess high potential as biological pretreatment agents by accelerating the depolymerization of the lignocellulosic matrix. This effect could reduce composting times and enable applications that favor its inclusion in circular economy frameworks. Full article

Wheat and rice are major sources of human nutrition worldwide. Solid-state fermentation (SSF) with lignocellulolytic mushrooms can enhance their nutritional value and increase their functional properties. However, this technology is not yet widely applied. In this work, whole wheat and brown rice hydrated to 60% were used as substrates for the edible mushroom Pleurotus ostreatus and the medicinal Ganoderma sessile, which were incubated for 14 days at 25 °C in the dark. The fermented substrate biomass was incorporated into standard sugar-snap cookie recipes, substituting 20% of the wheat flour. We evaluated the technological and nutritional properties of alternative fermented flours and cookies. Both the fermented flours and cookies exhibited increased soluble and total protein content, antioxidant power, and phenol content, indicating overall functional improvement. Fermented G. sessile flour also showed increased triterpenoid content. The physical quality of cookies remained within expected ranges, demonstrating the feasibility of the application. These results highlight the potential of SSF as a method for nutritional and functional enrichment of grains and extend the health benefits of mushrooms to populations relying on low-cost, grain-derived carbohydrates. Further studies on digestibility and in vivo activity of metabolites are needed to confirm the potential health benefits of fermented flours. Full article

Cremastra appendiculata, a rare medicinal orchid, has extremely low natural seed germination due to immature embryos and dense seed coats, impeding its conservation. Commensal germination with fungi is effective, but the action pathways remain unclear. This study combined morphological observation (scanning electron microscopy and section observation), physiological–biochemical detection (lignocellulolytic enzyme activities, nutrient/hormone contents, FTIR analysis) and transcriptomics to explore Coprinellus radians’ role in C. appendiculata seed germination, with commensal and non-commensal cultures on OMA medium set as experimental and control groups. Results showed C. radians significantly promoted C. appendiculata seed germination and protocorm development (superior to non-commensal conditions). Morphologically, C. radians hyphae invaded seed coats at 6 days post-inoculation; embryos broke through coats and formed apical meristems at 12 days, developing into peloton-containing protocorms at 25 days (breaking dormancy). Physiologically, C. radians secreted lignocellulolytic enzymes (laccase, cellulase, xylanase) to degrade coats, enhancing permeability and water uptake, while driving nutrient accumulation (starch, soluble sugars) and hormone balance. Transcriptomically, symbiosis activated carbon/energy metabolism genes, enriching starch-sucrose metabolism and glycolysis pathways. This study clarifies C. radians’ multi-dimensional action pathways in promoting C. appendiculata germination, providing support for rare orchid conservation. Full article

Straw returning, a core practice in conservation tillage, promotes sustainable intensification; however, it faces challenges such as inefficient decomposition, nutrient competition, and pathogen accumulation. To address these limitations, this study aimed to develop a multifunctional microbial consortium specifically designed for straw-incorporating cropping systems. The consortium comprises four Bacillus strains with complementary enzymatic systems, isolated from diverse ecological niches. It exhibited robust lignocellulolytic enzyme production, with manganese peroxidase (7709.33 U/L), laccase (450.65 U/L), endo-β-1,4-glucanase (154.67 U/mL), and filter paper activity (309.18 U/L). The consortium significantly enhanced rice straw degradation by 37.18% and increased nitrogen (N) release by 16.13% compared to the control. Moreover, the consortium exhibited a 67.56% inhibition rate against Magnaporthe oryzae and reduced both the incidence rate and disease index of leaf blast and panicle blast. Field trials revealed increases in the rice grain yield of 9.63% and 6.94% when applied alone and 6.75% and 5.18% when co-applied with straw residues. These findings highlight the multifunctional agricultural potential of the consortium and provide a sustainable strategy to overcome the limitations of straw-incorporating farming systems. Full article

The valorization of lignocellulosic biomass has been attracting interest in several industrial areas due to its potential to produce high-value-added compounds. Among these products, lignocellulosic enzymes stand out, capable of degrading biomass into fermentable polysaccharides, essential to produce second-generation ethanol and other bioproducts. The genus Pleurotus spp., a macrofungus with a high enzyme production capacity, has been consolidating itself as a promising alternative in the bioconversion of lignocellulosic residues. Contextually, this review explores for the first time the level of technology readiness associated with the production of enzymes by Pleurotus spp., in addition to addressing advances in patent filings and the role of these enzymes in the conversion of lignocellulosic biomass. Through a technological analysis based on a critical evaluation of 250 studies indexed in the database Scopus^® between 2015 and 2025, from which 16 studies were selected for a detailed and rigorous assessment of enzyme production by Pleurotus spp., it was observed that technological progress remains at the laboratory scale, in TRL 3 and 4, with few studies at the TRL 5 scale. In addition, the factors that may be affecting the increase in technological readiness of microbial enzyme production at pilot and industrial scales are discussed. The valorization of lignocellulosic biomass and the production of enzymes by macrofungi represent a promising path towards sustainability and cost reduction; however, significant challenges remain related to pilot-scale studies and increasing the level of technological maturity of these processes by Pleurotus spp., requiring further investigation of these processes to standardize and enable their industrial scale-up. Full article

The white-rot fungus Irpex lacteus is recognized for its strong ligninolytic and polysaccharide-degrading capacity, but the key advantages in degrading lignocellulose and the regulation of its enzyme systems remain poorly understood. In this study, we identified a rich repertoire of carbohydrate-active enzymes in the genome of I. lacteus QJ. Relative to other white-rot fungi, an expanded glycoside hydrolase gene family in I. lacteus QJ suggesting strong potential for lignocellulose degradation. When I. lacteus QJ was cultivated on glucose or wheat straw for 4 and 8 days, wheat straw strongly induced carbohydrate-active enzyme genes on day 4, while ligninolytic enzyme genes exhibited delayed upregulation on day 8. The cellobiose dehydrogenase plays an important role in the degradation processes. Its expression pattern is consistent with that of cellulase, and it can support peroxidase activity by providing H₂O₂. These findings reveal temporal coordination between polysaccharide- and lignin-degrading enzymes, providing new theoretical ideas for the application of I. lacteus during the degradation process. Our results not only improve the mechanistic understanding of fungal lignocellulose deconstruction but also inform strategies to enhance biological pretreatment of agricultural residues for biorefinery applications. Full article

Saprotrophic and lignocellulolytic fungi from tropical areas especially represent a promising yet relatively underexplored frontier for both taxonomy and applied research. This makes ex situ conservation through culture collections of paramount importance. Here, 10 lignocellulolytic strains isolated from the State of São Paulo (Brazil) and deposited in the Brazilian Culture Collection (now CCIBt) were identified through the ITS region. In order to prevent accidental losses, these strains have been shared with the collection of the University of Milano—Department of Food, Environmental and Nutritional Sciences (DeFENS), as well as the MicUNIPV Fungal Research Culture Collection—University of Pavia (Italy). Most of the fungal species in the examined set exhibit a neotropical distribution, while 3 out of 10 are nowadays recognized as subcosmopolitan despite their prevalence in the neotropical area. One holotropical, one cosmopolitan and one holarctic species are also present. Based on the literature, 8 out of the 10 characterized species are known to produce psilocybin (e.g., Psilocybe cubensis and Candolleomyces candolleanus) and/or enzymes with potential applications in environmental and medical biotechnology (e.g., Lentinus crinitus). All 10 strains were described for their micro- and macro-characteristics; their growth rate was evaluated and culture pictures provided. Taxonomic and nomenclatural controversies concerning Candolleomyces candolleanus, Cubamyces lactineus and Pycnoporus sanguineus are discussed. Full article

Flammulina filiformis, one of the most delicious and commercially important mushrooms, demonstrates remarkable adaptability to diverse agricultural wastes. However, it is unclear how different substrates affect the degradation of lignocellulosic biomass and the production of lignocellulolytic enzymes in F. filiformis. In this study, label-free comparative proteomic analysis of F. filiformis cultivated on sugarcane bagasse, cotton seed shells, corn cobs, and glucose substrates was conducted to identify degradation mechanism across various substrates. Label-free quantitative proteomics identified 1104 proteins. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis of protein expression differences were predominantly enriched in energy metabolism and carbohydrate metabolic pathways. Detailed characterization of carbohydrate-active enzymes among the identified proteins revealed glucanase (GH7, A0A067NSK0) as the key enzyme. F. filiformis secreted higher levels of cellulases and hemicellulases on sugarcane bagasse substrate. In the cotton seed shells substrate, multiple cellulases functioned collaboratively, while in the corn cobs substrate, glucanase predominated among the cellulases. These findings reveal the enzymatic strategies and metabolic flexibility of F. filiformis in lignocellulose utilization, providing novel insights for metabolic engineering applications in biotechnology. The study establishes a theoretical foundation for optimizing biomass conversion and developing innovative substrates using targeted enzyme systems. Full article

Pleurotus ostreatus is a mushroom species renowned for its abundant nutritional and medicinal properties. Nevertheless, the yield of its fruiting bodies has long remained at a standstill, making it arduous to achieve substantial improvements. Because the traditional composting approach for enhancing the yield of Pleurotus ostreatus has drawbacks such as a long duration and a high susceptibility to mold contamination, incorporating nutritional supplements into the culture medium of P. ostreatus has emerged as a relatively straightforward yet effective approach to enhancing its yield. This study was predicated on the roles of nucleotides and nucleosides in cellular metabolism and signal transduction. These substances were applied during the cultivation process of P. ostreatus to investigate their impact on the growth and nutritional composition of this mushroom. The findings of this study demonstrate that the supplementation of nucleotides and nucleosides not only improved the yield and biological efficiency of P. ostreatus but also increased its dietary fiber content and amino acids. Furthermore, this research has disclosed that nucleotides and nucleosides exert a notable influence on the lignocellulolytic enzyme system. This investigation provides a scientific foundation for the development of novel yields—enhancing agents for P. ostreatus and offering new insights into cultivation techniques for the progress of P. ostreatus cultivation techniques in both academic and practical arenas. Full article

The valorization of plant biomass is one of the main strategies for sustainable development. However, its use as energy, biofuels, fertilizers, value-added products, or even food is severely affected by the complexity of the plant cell wall. Therefore, the evaluation of fungi with high production of lignocellulolytic enzymes capable of efficiently degrading these substrates constitutes a viable, clean, and eco-friendly solution, allowing, for example, an increase in the digestibility and nutritional quality of alternative animal feed sources. For these reasons, the present study evaluated the ability of the mutant strain Trichodema viride M5-2 to improve the nutritional composition of the forage legumes Lablab purpureus and Mucuna pruriens through solid-state fermentation. Endo- and exoglucanase cellulolytic activity was assessed, as well as the effect of fermentation on the fiber’s physical properties and chemical composition. Molecular changes in the structure of plant fiber were analyzed using infrared spectroscopy. Increased production of the cellulolytic complex of the enzymes endoglucanase (3.29 IU/mL) and exoglucanase (0.64 IU/mL) was achieved in M. pruriens. The chemical composition showed an increase in true protein and a decrease in neutral fiber, hemicellulose, and cellulose, with a consequent improvement in nutritional quality. Fiber degradation was evident in the infrared spectrum with a significant decrease in the signals associated with cellulose and, to a lesser extent, with lignin. It can be concluded that the mutant strain T. viride M5-2 produced chemical, physical, and molecular changes in the fibrous and protein fractions of L. purpureus and M. pruriens through SSF, which improved their nutritional value as an alternative feed for animal nutrition. By promoting the use of this fungus, the nutritional quality of this source is increased through an effective and eco-friendly process, which contributes to mitigating the environmental impact of food production, in accordance with sustainability objectives and the need for more responsible agricultural practices. Full article