Search Results (204)

This study aimed to isolate and identify fungi capable of producing xylanases and cellulases. Thirty-eight fungal strains were isolated from decaying water hyacinth (WH), and two were selected based on their superior enzyme production under solid-state fermentation (SSF). The strains were identified through morphological, cultural, and molecular analyses as Aspergillus austwickii B6 and Trichoderma harzianum M7. Their ribosomal ITS sequences were deposited in GenBank under accession numbers PQ142799.1 for A. austwickii B6 and PQ007458.1 for T. harzianum M7. Enzyme production was evaluated under SSF using eight culture medium variants prepared with natural or pretreated biomasses of WH and sugarcane bagasse (SCB), combined with either NaNO₃ or (NH₄)₂SO₄ as nitrogen sources. The maximum xylanase and cellulase activities were 752 and 65 U/g dry matter (DM), respectively, for A. austwickii B6, and 1724 and 152 U/g DM for T. harzianum M7, when cultivated on a low-cost medium composed of pretreated WH, (NH₄)₂SO₄, and a simple mineral salt solution. These findings highlight the potential of locally isolated fungal strains and lignocellulosic residues as cost-effective substrates and inducers of xylanase and cellulase production under SSF and underscore the importance of WH pretreatment to enhance substrate availability and maximize enzyme yields. Full article

Sustainable development requires implementation of eco-friendly practices and a circular approach in both agricultural and industrial systems. This study evaluated winery flotation wastewater (WFW) as a cultivation substrate for Bacillus sp. 10/R isolated from grapevine rhizosphere for sustainable biostimulant production. The bacterial isolate was characterized by 16S rRNA sequencing and biochemical tests, showing the highest similarity with Bacillus mojavensis and Bacillus halotolerans. Plant growth-promoting traits were assessed via assays for hydrolytic enzymes, ACC (1-aminocyclopropane-1-carboxylate) deaminase, and IAA (indole acetic acid) production, as well as for phosphate solubilization. The isolate was cultivated in WFW, including monitoring of biomass growth, enzymatic activity, and substrate composition changes. The resulting cultivation broths based on WFW (WFW-CB) and nutrient broth (NB-CB) were tested as barley seed treatment at five dosages, using sterile media and water as controls. The results have displayed strong pectinase (EAI–enzyme activity index 2.79) and cellulase activity (2.33), moderate xylanase (1.75) and ACC deaminase activity (growth zone 54.67 ± 0.58 mm), and moderate IAA production (9.66 µg/mL). Biomass content has increased by two log units within 48 h (up to 9.06 log CFU/mL), with stable pectinase activity (~2.2 U/mL). Germination assays revealed that 10% WFW-CB and 50% WFW enhanced germination indices and biomass, whereas undiluted WFW and WFW-CB inhibited germination. These results indicate that WFW is a suitable substrate for Bacillus sp. 10/R cultivation, linking industrial wastewater valorization with plant biostimulant production in a circular economy framework. Full article

In recent years, in response to the demand for the livestock industry to enhance cattle production performance, scholarly inquiries have centered on elucidating the underlying mechanisms by which feed additives modulate rumen microenvironment and metabolic efficacy, thereby facilitating nutrient absorption and augmenting production performance in cattle. This study was undertaken to evaluate the impacts of surfactant polyacrylamide (PAM) supplementation on digestive processes, metabolic dynamics, and ruminal digestive enzyme activities in cattle. Four ruminally cannulated crossbred cows (~3 years, 350 kg, non-pregnant/lactating) were utilized in a 4 × 4 Latin square design. The animals were fed a basal diet supplemented with polyacrylamide (PAM) at concentrations of 0, 1.0, 2.0, and 6.0 g/kg across four 22-day experimental periods, each consisting of a 16-day adaptation phase and a 6-day sampling phase. Supplementation with polyacrylamide (PAM) at levels ranging from 1.0 to 6.0 g/kg of diet significantly increased voluntary dry matter intake (VFI) in cattle by a maximum of 13.7% (p < 0.05), with peak effects at 2.0 g/kg. The digestibility of dry matter, crude protein, cellulose, and energy was significantly improved (p < 0.05 to p < 0.01), reaching maximum increases by 12.6%, 12.8%, 17.5%, and 11.7%, respectively. Nitrogen, calcium, and phosphorus retention increased substantially (p < 0.01 to p < 0.05), with calcium retention showing the highest improvement (55.7%). Rumen cellulase activities (endocellulase, exocellulase, cellobiase, and xylanase) were significantly enhanced (p < 0.01), peaking at 37.3% for cellobiase. However, pectase, amylase, and protease activities remained unaffected. Optimal benefits were observed at 2.0 g/kg PAM, highlighting its potential to improve feed efficiency and nutrient utilization in cattle. Full article

The resource utilization of peach wood as agricultural waste holds significant importance for the sustainable development of the edible fungi industry, yet its regulatory effects on the physiology and safety of Lentinula edodes (L. edodes) remain unclear. This study selected four L. edodes (F2, 0912, N5, and 215) and systematically analyzed their cultivation adaptability across five peach wood substrate proportions (0%, 20%, 40%, 60%, and 80%). Results indicated that while high peach wood proportions inhibited laccase activity and delayed mycelial growth, high carboxymethyl cellulase and xylanase activity formed a critical compensatory effect, ultimately enhancing total yield. Peach wood improved production through strain-specific mechanisms. F2 increased via single mushroom weight gain, while N5 relied on xylanase-driven primordia differentiation to boost mushroom numbers. Adding peach wood significantly increased crude protein, crude lipid, and total polysaccharide in F2, maintaining normal agronomic traits and increasing secondary mushroom proportion. Safety risks focused on arsenic accumulation, with 80% peach wood causing F2 to exceed control levels, albeit remaining far below the national standards. This study is among the first to elucidate peach wood’s temporal enzyme regulation for the maintenance of L. edodes yield. Future optimization through peach wood pretreatment and low arsenic strain selection could provide technical support for the high value utilization of agricultural waste. Full article

Calocybe gambosa (Fr.) Donk is an edible mushroom, highly appreciated especially in Italy. It forms fairy rings (FRs) characterized by a zone of dead vegetation corresponding to the underground-extending mycelial front, followed by a “greener belt” where vegetation is thriving. To better understand this particular phenomenon, the effect of C. gambosa mycelium on plants were studied both in situ, across different zones of FRs (external area—EX, fungal front—FF, greener belt—GB, internal area—IN) of three fairy rings, and ex situ on Poa trivialis L. Plant community analysis revealed significant changes in plant species composition across the zones, characterized by a decline in diversity and a vegetation shift, from dicotyledons to monocotyledons, progressing from the EX toward the IN, where vegetation gradually begins to reestablish its original composition. Molecular and morphological analyses showed the endophytic colonization of C. gambosa mycelium within the herbaceous plants growing at the FF. Ex situ studies indicated pathogenic behavior of C. gambosa. After root colonization, it caused growth reduction in P. trivialis plants (79% reduction in root length, 76% reduction in leaf length), leaf yellowing, decreased photosynthetic pigments, and root necrosis. The cellulase (endo-1,4-β-glucanase), xylanase, polygalacturonase, and polymethylgalacturonase enzymatic activities of C. gambosa support its pathogenic effects. Conversely, volatile organic compounds (VOCs) produced by C. gambosa mycelium stimulated shoot development in P. trivialis (17% increase in shoot length), which accounts for the formation of the flourishing vegetation zone behind the FF. In contrast, soluble substances produced by C. gambosa mycelium did not affect the growth of P. trivialis. Our results suggest that C. gambosa plays a dual ecological role in regulating plant community dynamics within FRs: it acts as a pathogen by colonizing herbaceous plant roots and, at the same time, promotes vegetation growth through VOC production. Full article

Native yeasts are a promising microbial resource for the development of sustainable biorefineries. In this study, we isolated 30 yeast strains from soil, decaying wood, and tree bark in a preserved Araucaria Forest in Southern Brazil and characterized them phenotypically and taxonomically. All strains were able to grow on glucose, xylose, and cellobiose, and 50% of them could metabolize arabinose. Several isolates showed high growth rates on xylose (up to 0.47 h⁻¹) and cellobiose (up to 0.45 h⁻¹). Notably, 19 strains (63% of the analyzed yeasts) exhibited xylanase activity at 50 °C (up to 156.84 U/mL), and four strains (13%) showed significant cellulase production. β-Glucosidase activities were particularly high in permeabilized cells of CHAP-258, CHAP-277, and CHAP-278 (up to 584.33 U/mg DCW), with kinetic parameters indicating high enzymatic performance. Twelve strains (40% of the total) were classified as oleaginous, and three (10%) displayed both lipogenic and esterase activity. Lipase activity against p-nitrophenyl palmitate (pNPP) reached 55.55 U/mL in CHAP-260. Taxonomic identification revealed representatives of seven genera, including Meyerozyma, Papiliotrema, Scheffersomyces, and Sugiyamaella, with potential for biotechnological use. Overall, the biochemical diversity observed highlights the value of native yeasts from Araucaria Forests as biocatalysts for lignocellulose-based bioprocesses, particularly due to their ability to grow on pentoses, secrete hydrolytic enzymes, and accumulate lipids. Full article

This study evaluates the effects of two Trichoderma reesei exogenous fibrolytic enzyme (EFE) preparations on the taxonomic profile, diversity, relative abundance, and population shifts of three ruminal bacteria fractions of lactating cows: free-floating (LIQ), weakly (AS), and tightly (SOL) feed-adhered. Three lactating cows were fed three EFE treatments in a 3 × 3 Latin square design: one control (CON) without enzymes, a cellulase/xylanase mix (MIX), and a high-xylanase treatment (XYL). Rumen contents were collected, and bacteria were extracted from the three ruminal content fractions for next-generation sequencing analysis. Alpha diversity was higher in XYL compared to CON. However, no EFE effect was observed on beta diversity. The relative abundance (RA) of the family Prevotellaceae increased, while that of Ruminococcaceae and Rikenellaceae decreased in XYL compared to MIX and CON. The bacterial community structure (beta diversity) of LIQ was differentiated from that of SOL and AS (p = 0.03), but no effects of fraction were observed on alpha diversity. Lachnospiraceae RA was greater in SOL, followed by AS, and lower in LIQ (p < 0.001), while Spirochaetaceae RA was greater in SOL and AS compared to LIQ (p = 0.003). The effects of EFE supplementation on rumen bacterial RA were independent of the ruminal content fraction. Full article

Microbial enzymes, due to their efficiency, specificity, and sustainability, are central to innovative biotechnological strategies aimed at optimizing industrial processes such as winemaking. In this study, the potential of Aureobasidium pullulans m11-2, a native dimorphic fungus from the wine ecosystem, was evaluated as a source of hydrolytic enzymes capable of degrading grape cell wall polysaccharides. The strain was identified at the molecular level and characterised in terms of its morphology. To maximise enzyme production, various culture media were tested. Among the concentrations tested, the optimal levels of glucose and pectin were 1 g L⁻¹ and 10 g L⁻¹, respectively. The partially constitutive and inducible nature of the various polysaccharidase activities (pectinases, cellulases, and xylanases) was confirmed. The effect of grape skins (a winemaking by-product) on microbial growth and enzyme synthesis was evaluated, achieving a pectinase activity of 0.622 U mL⁻¹ when combined with 1 g L⁻¹ of glucose. Maximum enzyme yields were detected during the exponential growth phase in both citrus pectin and grape skin media, suggesting favorable conditions for continuous bioprocessing. These results confirm that A. pullulans m11-2 is an interesting microbial option for producing polysaccharidases that can be adapted to sustainable production systems. Full article

The use of exogenous enzymes in the nutrition of dairy cows is an innovative and efficient strategy to maximize productivity and milk quality, with positive applications in the economic and environmental aspects of dairy farming. Therefore, the objective of this study was to evaluate whether the addition of a blend of exogenous enzymes to the diet of lactating Jersey cows has a positive effect on productive performance, milk quality, animal health, ruminal environment, and digestibility. Twenty-one primiparous Jersey cows, with 210 days in lactation (DL), were used. The exogenous enzymes used were blends containing mainly protease, in addition to cellulase, xylanase, and beta-glucanase. The animals were divided into three groups with seven replicates per group (each animal being the experimental unit), as follows: Control (T-0), basal diet without enzyme addition; Treatment (T-80), animals fed enzymes in the diet at a daily dose of 80 mg per kg of dry matter (DM); Treatment (T-160), animals fed enzymes in the diet at a daily dose of 160 mg per kg of DM. The study lasted 84 days, during which higher milk production was observed in the treated groups (T-80 and T-160) compared to the control group (p = 0.04). When calculating feed efficiency from days 1 to 84, greater efficiency was observed in both groups that received the blend compared to the control (p = 0.05). In the centesimal composition of the milk, it was observed that the percentage of protein in the milk of the T-160 group was higher compared to the control group (p = 0.03). The effect of the enzymes was verified for butyric (p = 0.05) and palmitic (p = 0.05) fatty acids. We also observed the effect of the enzyme blend on the amount of volatile fatty acids (VFAs), which were higher in the ruminal fluid of cows that received the enzymes (p = 0.01). Cows that consumed enzymes showed a higher apparent digestibility coefficient of crude protein (p = 0.01). In vitro, the main result is related to lower gas production in 24 and 48 h at T-160. We concluded that the use of a blend of exogenous enzymes in the diet of lactating Jersey cows was able to increase milk production in these animals, resulting in greater feed efficiency and also an increase in milk protein content, positively modulating the fatty acid profile in the rumen and improving the apparent digestibility of nutrients. Full article

Using wheat flour milling (WFM) co-products in pig diets may reduce feed cost. Still, energy digestibility is lower for WFM co-products than for feed grains. Inadequate information exists about their fermentation characteristics and the relationship between digestible energy (DE) value and chemical characteristics or in vitro energy digestibility. The objectives were to (1) determine the chemical characteristics, in vitro and in vivo DE values, and energy digestibility of WFM co-products in growing pigs; (2) determine their in vitro microbial fermentation characteristics, and (3) establish relationships between in vivo DE value of WFM co-products and their chemical composition, fermentation characteristics, or in vitro digestibility values. Across Canada, 94 WFM co-products were sampled and characterized for their chemical composition and in vitro dry matter (DM) and energy digestibility using pepsin, pancreatin, and a multi-enzyme complex containing arabinase, β-glucanase, hemicellulase, xylanase, and cellulase. The in vivo energy, DM digestibility and DE value of 9 WFM co-products (2 shorts, 5 millrun, 1 middling, and 1 bran) were determined using a corn-based diet and 40 growing pigs in two periods to obtain 8 observations per diet. After in vitro digestion, the 9 WFM co-product samples were subjected to microbial fermentation using fresh fecal inoculum in a cumulative gas-production technique. The WFM co-products had a high content of crude fiber (up to 7.9% in shorts, 9.9% in millrun, 7.1% in middlings, and 12.0% in bran) and crude protein (CP; up to 27.8% in shorts, 20.0% in millrun, 22.1% in middlings, 15.9% in bran). The DE values ranged from 2.84 to 3.74 Mcal/kg DM among WFM co-products. Among chemical characteristics, neutral detergent fiber was the best predictor (R² = 0.81) for in vivo DE value, followed by crude fiber (R² = 0.78), and acid detergent fiber (R² = 0.72). The in vitro DE values predicted (R² = 0.80) in vivo DE values of 9 WFM co-products. Based on principal component analysis, total gas and short-chain fatty acid production varied among WFM co-products and was associated with the CP content of WFM co-products. In conclusion, WFM co-products contain high crude protein and have a high DE value for growing pigs but vary substantially in nutritional value. Full article

Mesocotyl elongation is the key determinant of deep-sowing tolerance in maize. Sowing at an appropriate depth allows the seedling to exploit water and nutrients stored in deeper soil layers, thereby enhancing its ability to withstand drought and other abiotic stresses. Mesocotyl elongation is regulated by the phytohormones brassinosteroid (BR), auxin (IAA), gibberellin (GA), and reactive oxygen species (ROS). However, whether and how BR coordinates IAA, GA, and ROS to control mesocotyl elongation in maize remains unclear. Here, we demonstrated that BRs orchestrate ROS, IAA, and GA signaling to remodel cell-wall metabolism in mesocotyl cells, promote cell elongation, and, consequently, strengthen deep-sowing tolerance. BR promoted mesocotyl elongation through multiple routes: (1) decreasing the contents of cell-wall components (hemicellulose, cellulose, and pectin); (2) activating cell-wall-loosening enzymes (cellulase, pectinase, and acidic xylanase); and (3) disturbing ROS homeostasis by elevating superoxide dismutase (SOD) activity. Combined treatments of BR with either IAA or GA further enhanced mesocotyl elongation in a concentration-dependent manner. In deep-sowing trials (15 cm), application of BR alone or in combination with IAA or GA markedly increased mesocotyl length and emergence rate, thereby improving deep-sowing tolerance. Our work indicated that BR integrated ROS, IAA, and GA signals to restructure the cell wall and derived mesocotyl cell elongation, providing both theoretical insights and practical strategies for breeding maize varieties with enhanced deep-sowing tolerance. Full article

This study aimed to evaluate solid-state fermentation (SSF) as a sustainable approach for the simultaneous detoxification of olive pomace (OP) and the production of industrially relevant enzymes. OP, a semisolid byproduct of olive oil extraction, is rich in lignocellulose and phenolic compounds, which limit its direct reuse due to phytotoxicity. A native strain of Aspergillus sp., isolated from OP, was employed as the biological agent, while grape pomace (GP) was added as a co-substrate to enhance substrate structure. Fermentations were conducted at two scales, Petri dishes (20 g) and a fixed-bed bioreactor (FBR, 2 kg), under controlled conditions (25 °C, 7 days). Key parameters monitored included dry and wet weight loss, pH, color, phenolic content, and enzymatic activity. Significant reductions in color and polyphenol content were achieved, reaching 68% in Petri dishes and 88.1% in the FBR, respectively. In the FBR, simultaneous monitoring of dry and wet weight loss enabled the estimation of fungal biotransformation, revealing a hysteresis phenomenon not previously reported in SSF studies. Enzymes such as xylanase, endopolygalacturonase, cellulase, and tannase exhibited peak activities between 150 and 180 h, with maximum values of 424.6 U·g⁻¹, 153.6 U·g⁻¹, 67.43 U·g⁻¹, and 6.72 U·g⁻¹, respectively. The experimental data for weight loss, enzyme production, and phenolic reduction were accurately described by logistic and first-order models. These findings demonstrate the high metabolic efficiency of the fungal isolate under SSF conditions and support the feasibility of scaling up this process. The proposed strategy offers a low-cost and sustainable solution for OP valorization, aligning with circular economy principles by transforming agro-industrial residues into valuable bioproducts. Full article

This study investigated the individual and combined effects of α-amylase (6 and 10 ppm), xylanase (70 and 120 ppm), and cellulase (35 and 60 ppm) on the physicochemical and nutritional properties of Chinese steamed buns (CSBs) supplemented with 15% buckwheat flour. The addition of individual enzymes did not significantly affect the volume or texture of the buckwheat-enriched CSBs, although it increased the crumb moisture content and porosity. In contrast, enzyme combinations can improve specific volume and reduce hardness. The enzyme combination (α-amylase 6 ppm, xylanase 70 ppm, and cellulase 60 ppm) yielded the highest specific volume (2.50 mL/g) and the lowest hardness (271.46 g). Regarding chemical properties, individual enzymes had minimal impact, while the combined treatment (6, 70, 60 ppm) decreased starch and dietary fiber content. For nutritional properties, the glycemic response of the CSBs varied depending on the concentration of the enzyme combination used. Full article

Thermostable cellulases and xylanases have broad acceptance in food, feed, paper and pulp, and bioconversion of lignocellulosics. Thermophilic fungi serve as an excellent source of thermostable enzymes. This study characterized four endo-β-1,4-glucanases (two glycoside hydrolase (GH) family 5 and two GH7 members) and four endo-β-1,4-xylanases (two GH10 and two GH11 members) from thermophilic fungus Thielavia terrestris, along with one GH10 endo-β-1,4-xylanase each from thermophilic fungus Chaetomium thermophilum and mesophilic fungus Chaetomium globosum. Comparative analysis was conducted against three previously reported GH10 endoxylanases: two thermostable enzymes from the thermophilic fungus Humicola insolens and thermophilic bacterium Halalkalibacterium halodurans, and one mesophilic enzyme from model fungus Neurospora crassa. The GH10 xylanase TtXyn10C (Thite_2118148; UniProt G2R8T7) from T. terrestris demonstrated high thermostability and activity, with an optimal temperature of 80–85 °C. It retained over 60% of its activity after 2 h at 70 °C, maintained approximately 30% activity after 15 min at 80 °C, and showed nearly complete stability following 1 min of exposure to 95 °C. TtXyn10C exhibited specific activity toward beechwood xylan (1130 ± 15 U/mg) that exceeded xylanases from H. insolens and H. halodurans while being comparable to N. crassa xylanase activity. Furthermore, TtXyn10C maintained stability across a pH range of 3–9 and resisted trypsin digestion, indicating its broad applicability. The study expands understanding of enzymes from thermophilic fungi. The discovery of the TtXyn10C offers a new model for investigating the high activity-stability trade-off and structure-activity relationships critical for industrial enzymes. Full article

Rumen microbiota is crucial for cellulose degradation and nutrient metabolism in ruminants. Different feeding systems like grazing and housed feeding can significantly impact it. Mongolian cattle show unique cellulose degradation ability, but functional changes under different feeding conditions are unclear. This study aims to investigate the effects of grazing and housed feeding on rumen microbiota, cellulose degradation, and metabolism in Mongolian cattle. In a 90-day trial, 12 female Mongolian cattle were divided into grazing (F group) and housed feeding (S group). Rumen samples were collected to analyze fermentation parameters, enzyme activities, microbiomes, and metabolomes. The F group had higher acetate, cellulase, xylanase, and β-glucosidase activities (p < 0.05). Bacteroidota and Prevotella were more abundant (p < 0.05), while Firmicutes and Ruminococcus were less abundant (p < 0.05) in the F group. Carbohydrate metabolic pathways and CAZymes (GH2, GH10) were upregulated in the F group, while the S group had enriched purine metabolic pathways and CAZyme (GH31). A total of 64 differential metabolites were found, with subaphylline upregulated in the F group and L-arogenate in the S group (p < 0.05). Grazing increased cellulose degradation and subaphylline production in Mongolian cattle, while housed feeding improved starch utilization efficiency and fat synthesis. These findings provide a basis for optimizing feeding strategies and improving fibrous feed resource utilization in Mongolian cattle. Full article