Search Results (606)

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

Hyaluronidase is a hydrolytic enzyme that cleaves β-1,4-glycosidic linkages in high-molecular-weight hyaluronic acid, generating low-molecular-weight oligosaccharides with enhanced biological functions. These products exhibit immunomodulatory, antioxidant, and tissue-regenerative properties, making them valuable in pharmaceutical, cosmetic, and functional food applications. However, most commercial hyaluronidases originate from pathogenic bacteria or recombinant hosts, raising concerns over their biosafety and regulatory acceptance, particularly in food-grade applications. In this study, we report the isolation and characterization of a novel non-pathogenic soil bacterium, Paenibacillus residui BSSK58, which produces an extracellular hyaluronidase. Whole-genome sequencing revealed the absence of known virulence factors and antibiotic resistance genes. Phenotypic safety evaluations confirmed that there was no hemolytic activity, biogenic amine production, or cytotoxicity against human intestinal epithelial cell lines (Caco-2 and HT-29). The purified BSSK58 hyaluronidase exhibited a molecular weight of approximately 170 kDa, with optimal activity at pH 8.0–9.0 and 50 °C. The enzyme showed broad substrate specificity toward hyaluronic acid, chondroitin sulfate, and alginate, and its depolymerizing activity was confirmed using gel permeation chromatography. Furthermore, a 13-week oral repeated-dose toxicity study under Good Laboratory Practice conditions demonstrated no adverse effects. These findings support the use of BSSK58 hyaluronidase as a safe, non-recombinant biocatalyst suitable for industrial applications under regulatory-compliant frameworks. Full article

Talaromyces pinophilus is a filamentous fungus with notable lignocellulose-degrading capacity based on enzyme activities and protein secretion potential, making it a compelling candidate for industrial biotechnology applications. In this study, we present the genomic characterization of the highly cellulolytic strain Y117, a domesticated variant of T. pinophilus, based on whole-genome sequencing and comparative genomic analysis with eleven related strains. Comprehensive analysis of CAZymes, transcription factors, and secondary metabolite diversity in T. pinophilus strains revealed that the exceptional lignocellulose degradation capacity of Y117 is driven by its unique genomic architecture. Key genomic features that distinguish Y117 include (1) significant expansion of glycoside hydrolase (GH) and carbohydrate-binding module (CBM) families, (2) loss of fungal-RiPP-like clusters, and (3) absence of the developmental regulator BrlA. These genomic adaptations could indicate a metabolic trade-off favoring hydrolytic enzyme production over secondary metabolism and sporulation. Our findings provide fundamental insights into fungal lignocellulose degradation mechanisms while establishing Y117 as a promising chassis for metabolic engineering applications in industrial enzyme production and heterologous protein expression. Full article

The citrus processing industry is an economically important agro-industrial sector worldwide; however, it produces significant amounts of waste annually. The biorefinery concept and the recovery of bio-based materials from agro-industrial residues, including citrus processing waste, are emphasized in the European Green Deal, reflecting the EU’s commitment to fostering circularity. Biotreatment of citrus processing waste, including bioconversion into biomethane, biohydrogen, bioethanol and biodiesel, has been applied to valorize biomass for energy recovery. It can also be composted into a valuable soil conditioners and fertilizers, while raw and fermented citrus residues may exhibit phytoprotective activity. Citrus-derived residues can be converted into materials such as nanoparticles with adsorptive capacity for heavy metals and recalcitrant organic pollutants, and materials with antimicrobial properties against various microbial pathogens, or the potential to remove antibiotic-resistance genes (ARGs) from wastewater. Indeed, citrus residues are an ideal source of industrial biomolecules, like pectin, and the recovery of bioactive compounds with added value in food processing industry. Citrus processing waste can also serve as a source for isolating specialized microbial starter cultures or as a substrate for the growth of bioplastic-producing microorganisms. Solid-state fermentation of citrus residues can enhance the production of hydrolytic enzymes, with applications in food and environmental technology, as well as in animal feed. Certain fermented products also exhibit antioxidant properties. Citrus processing waste may be used as alternative feedstuff that potentially improves the oxidative stability and quality of animal products. Full article

This study investigates the Enzyme Biofilm Method (EBM), a biological wastewater treatment technology previously developed by the authors. EBM employs microbial-derived hydrolytic enzyme groups in the initial treatment stage to break down high-molecular-weight organic matter—such as starch, proteins, and fats—into low-molecular-weight compounds. These compounds enhance the growth of native microorganisms, promoting biofilm formation on carriers and improving treatment efficiency. Over the past decade, EBM has been practically applied in food factory wastewater facilities handling high organic loads. The enzyme groups used in EBM are derived from cultures of Bacillus mojavensis, Saccharomyces cariocanus, and Lacticaseibacillus paracasei. To clarify the system’s mechanism and ensure its practical viability, this study focused on starch—a prevalent and recalcitrant component of food wastewater—using two evaluation approaches. Verification 1: Field testing at a starch factory showed that adding enzyme groups to the equalization tank effectively reduced biological oxygen demand (BOD) through starch degradation. Verification 2: Laboratory experiments confirmed that the enzyme groups possess both amylase and maltase activities, sequentially breaking down starch into glucose. The resulting glucose supports microbial growth, facilitating biofilm formation and BOD reduction. These findings confirm EBM’s potential as a sustainable and effective solution for treating high-strength food industry wastewater. Full article

Background/Objectives: Actinobacteria are one of the largest bacterial phyla. These microbes produce bioactive compounds, such as antifungals, antibiotics, immunological modulators, and anti-tumor agents. Studies on actinobacteria isolated from the Brazilian Savannah biome (Cerrado) are scarce and mostly address metagenomics or the search for hydrolytic enzyme-producing microbes. Solanum lycocarpum (lobeira) is a tree widely employed in regional gastronomy and pharmacopeia in Central Brazil. Methods: In this work, 60 actinobacteria isolates were purified from the rhizosphere of S. lycocarpum. Eight Streptomyces spp. isolates were selected for in vitro antifungal activity against Cryptococcus neoformans H99, the C. neoformans 89-610 fluconazole-tolerant strain, C. gattii NIH198, Candida albicans, C. glabrata, and C. parapsilosis. The ability of the aqueous extracts of the isolates to induce the in vitro secretion of tumor necrosis factor (TNF-α), nitric oxide (NO), interleukin-6 (IL-6), and IL-10 by murine macrophages was also evaluated. Results: All extracts showed antifungal activity against at least two yeast species. Streptomyces spp. LAP11, LDB2, and LDB17 inhibited C. neoformans growth by 40–93%. Most extracts (except LAP2) also inhibited C. gattii. None inhibited C. albicans, but all inhibited C. glabrata (40–90%). Streptomyces sp. LAP8 extract increased nitric oxide production by approximately 347-fold in murine macrophages, while LDB11 extract suppressed LPS-induced TNF-α production by 70% and simultaneously increased IL-10 secretion, suggesting immunosuppressive potential. Conclusions: The results revealed that Cerrado actinobacteria-derived aqueous extracts are potential sources of antifungal and immunomodulatory biocompounds. Full article

The sugarcane industry plays a crucial economic role worldwide, with sucrose and ethanol as its main products. However, its processing generates large volumes of by-products—such as bagasse, molasses, vinasse, and straw—that contain valuable components for biotechnological valorization. This review integrates approximately 100 original research articles published in JCR-indexed journals between 2015 and 2025, of which over 50% focus specifically on sugarcane-derived agroindustrial residues. The biotechnological approaches discussed include submerged fermentation, solid-state fermentation, enzymatic biocatalysis, and anaerobic digestion, highlighting their potential for the production of biofuels, enzymes, and high-value bioproducts. In addition to identifying current advances, this review addresses key technical challenges such as (i) the need for efficient pretreatment to release fermentable sugars from lignocellulosic biomass; (ii) the compositional variability of by-products like vinasse and molasses; (iii) the generation of metabolic inhibitors—such as furfural and hydroxymethylfurfural—during thermochemical processes; and (iv) the high costs related to inputs like hydrolytic enzymes. Special attention is given to detoxification strategies for inhibitory compounds and to the integration of multifunctional processes to improve overall system efficiency. The final section outlines emerging trends (2024–2025) such as the use of CRISPR-engineered microbial consortia, advanced pretreatments, and immobilization systems to enhance the productivity and sustainability of bioprocesses. In conclusion, the valorization of sugarcane by-products through biotechnology not only contributes to waste reduction but also supports circular economy principles and the development of sustainable production models. Full article

Fungal lytic polysaccharide monooxygenases (LPMOs) have revolutionized the field of biomass degradation by introducing an oxidative mechanism that complements traditional hydrolytic enzymes. These copper-dependent enzymes catalyze the cleavage of glycosidic bonds in recalcitrant polysaccharides such as cellulose, hemicellulose, and chitin, through the activation of molecular oxygen (O₂) or hydrogen peroxide (H₂O₂). Their catalytic versatility is intricately modulated by structural features, including the histidine brace active site, surface-binding loops, and, in some cases, appended carbohydrate-binding modules (CBMs). The oxidation pattern, whether at the C1, C4, or both positions, is dictated by subtle variations in loop architecture, amino acid microenvironments, and substrate interactions. LPMOs are embedded in a highly synergistic fungal enzymatic system, working alongside cellulases, hemicellulases, lignin-modifying enzymes, and oxidoreductases to enable efficient lignocellulose decomposition. Industrial applications of fungal LPMOs are rapidly expanding, with key roles in second-generation biofuels, biorefineries, textile processing, food and feed industries, and the development of sustainable biomaterials. Recent advances in genome mining, protein engineering, and heterologous expression are accelerating the discovery of novel LPMOs with improved functionalities. Understanding the balance between O₂- and H₂O₂-driven mechanisms remains critical for optimizing their catalytic efficiency while mitigating oxidative inactivation. As the demand for sustainable biotechnological solutions grows, this narrative review highlights how fungal LPMOs function as indispensable biocatalysts for the future of the Circular Bioeconomy and green industrial processes. Full article

The majority of studies of fungal utilization of chitosan are associated with the production of a specific enzyme, chitosanase, which catalyzes the hydrolytic cleavage of the macrochain. In our opinion, the development of approaches to obtaining materials with new functional properties based on non-destructive chitosan transformation by living organisms and their enzyme systems is promising. This study was conducted using a wide range of classical and modern methods of microbiology, biochemistry, and physical chemistry. The ability of the ascomycete Fusarium oxysporum Schltdl. to modify films of chitosan with average-viscosity molecular weights of 200, 450, and 530 kDa was discovered. F. oxysporum was shown to use chitosan as the sole source of carbon/energy and actively overgrew films without deformations and signs of integrity loss. Scanning electron microscopy (SEM) recorded an increase in the porosity of film substrates. An analysis of the FTIR spectra revealed the occurrence of oxidation processes and crosslinking of macrochains without breaking β-(1,4)-glycosidic bonds. After F. oxysporum growth, the resistance of the films to mechanical dispersion and the degree of ordering of the polymer structure increased, while their solubility in the acetate buffer with pH 4.4 and sorption capacity for Fe²⁺ and Cu²⁺ decreased. Elemental analysis revealed a decrease in the nitrogen content in chitosan, which may indicate its inclusion into the fungal metabolism. The film transformation was accompanied by the production of extracellular hydrolase (different from chitosanase) and peroxidase, as well as biosurfactants. The results obtained indicate a specific mechanism of aminopolysaccharide transformation by F. oxysporum. Although the biochemical mechanisms of action remain to be analyzed in detail, the results obtained create new ways of using fungi and show the potential for the use of Fusarium and/or its extracellular enzymes for the formation of chitosan-containing materials with the required range of functional properties and qualities for biotechnological applications. Full article

Fungal communities in high plateau ecosystems remain understudied despite their crucial roles in soil ecosystems, and yeasts inhabiting extreme regions have potential for industrial and biotechnological applications. We studied the fungal diversity in soils across 14 Chilean Altiplano sites using amplicon-based metagenomics and isolation of yeasts to assess their growth under various conditions and hydrolytic enzyme secretion. Using the metagenomic approach, the Ascomycota and Basidiomycota phyla were found to be the most abundant (85% and 8%, respectively). Unclassified families and genera prevailed at six and ten sites, respectively. At the other sites, the most abundant families included Cladosporiaceae, Teratosphaeriaceae, and Sporormiaceae, and the genera Oleoguttula, Coniochaeta, and Peziza. Biodiversity indices did not correlate with the soil’s geographic origin, organic matter content, humidity, or pH. Most isolated yeasts belong to the Naganishia, Holtermanniella, and Vishniacozyma genera, growing at temperatures ranging from 4 °C to 26 °C. Most isolates could use glucose, sucrose, and maltose as carbon sources and exhibited amylase, esterase, pectinase, and protease activities at 30 °C and below. Our results indicate that the evaluated soil physicochemical parameters do not explain the fungal distribution in the Altiplano and highlight the region as a reservoir of unknown fungi, including yeasts with industrially relevant enzymes. Full article

The objective of this study was the evaluation of fungal solid-state fermentation (SSF) for the production of alginate lyase and extraction of uronic acids from Sargassum sp. For this purpose, the fungi Trichoderma asperellum, Aspergillus oryzae, and Rhizopus oryzae were applied (alone or combined) to Sargassum sp. biomass through SSF (10⁷ spores g_biomass⁻¹, 30 °C, and 7 days of treatment). In general, individual SSF with all three fungi degraded the biomass, achieving a marked synergy in the production of cellulase, laminarinase, and alginate lyase activities (especially for the last one). Trichoderma was the most efficient species in producing laminarinase, whereas Rhizophus was the best option for producing alginate lyase. However, when dual combinations were tested, the maximal values of alginate lyase activities were reached (13.4 ± 0.2 IU g_biomass⁻¹ for Aspergillus oryzae and Rhizopus oryzae). Remarkably, uronic acids were the main monomeric units from algal biomass solubilization, achieving a maximum yield of 14.4 mg_uronic g_biomass⁻¹, with the A + R condition being a feasible, eco-friendly alternative to chemical extraction of this monomer. Additionally, the application of all the fungal pretreatments drastically decreased the total phenolic content (TPC) in the biomass from 369 mg L⁻¹ to values around 44–84 mg L⁻¹, minimizing the inhibition for possible subsequent biological processes in which the residual solid can be used. Full article

Stipagrostis pennata is an important plant in desert ecosystems. Its seed-endophytic bacteria may play a critical role in plant growth and environmental adaptation processes. This study systematically analyzed the community composition and potential plant growth-promoting (PGP) functions of seed-endophytic bacteria associated with S. pennata. The results showed that while the overall diversity of bacterial communities from different sampling sites was similar, significant differences were observed in specific functional genes and species abundances. Nine endophytic bacterial strains were isolated from the seeds, among which Bacillus altitudinis strain L7 exhibited phosphorus solubilizing capabilities, nitrogen fixing, IAA production, siderophore generation, and multi-hydrolytic enzyme activities. Additionally, the genomic sequencing of L7 revealed the key genes involved in plant growth promotion and environmental adaptation, including Na⁺ efflux systems, K⁺ transport systems, compatible solute synthesis genes, and the gene clusters associated with nitrogen metabolism, IAA synthesis, phosphate solubilization, and siderophore synthesis. Strain L7 exhibits salt and osmotic stress tolerance while promoting plant growth, providing a promising candidate for desert microbial resource utilization and plant biostimulant development. Full article

Plant subtilases, as hydrolytic enzymes, contribute to certain plant stress response pathways by cleaving precursor proteins into active peptides or through other less well-characterized mechanisms. Phytaspases represent a specific subgroup of subtilases, and their participation in rapid stress responses, particularly to herbivory attacks and drought, is already well established, in contrast to their poorly understood role in long-term responses. This study investigated the involvement of phytaspase NbSBT1.9-2 in the long-term stress responses of Nicotiana benthamiana. Plants were subjected to either mild to severe mechanical wounding or drought stress, followed by the detection of phytaspase activity and gene expression in the leaf tissue. The results revealed a distinct involvement of phytaspase in the wounding response, showing increased activity and upregulated expression correlated with the extent and recurrence of wounding. In contrast, no significant change in phytaspase activity was observed in the leaves under drought, alongside salinity and heat stress conditions. Consequently, phytaspase association with the long-term response to mechanical injury was demonstrated using N. benthamiana as a model organism. Full article

Anaerobic acidogenic fermentation presents a promising approach for sustainable carbon emission mitigation in livestock waste management, addressing critical environmental challenges in agriculture. This study systematically investigated the synergistic effects of composting-assisted pretreatment coupled with micro-aeration and methanogenesis suppression to enhance volatile fatty acid (VFA) production from swine manure supplemented with wheat straw, valorizing agricultural waste while reducing greenhouse gas emissions. The experimental protocol involved sequential optimization of pretreatment conditions (12 h composting followed by 10 min thermal pretreatment at 85 °C), operational parameters (300 mL micro-aeration and 30 mmol/L 2-bromoethanesulfonate (BES) supplementation), and their synergistic integration. The combined strategy achieved peak VFA production (5895.92 mg/L, p < 0.05), with butyric acid constituting the dominant fraction (2004.42 mg/L, p < 0.05). Enzymatic analysis demonstrated significantly higher activities of key hydrolytic enzymes (protease, α-glucosidase) and acidogenic enzymes (butyrate kinase, acetate kinase) in the synergistic treatment group compared to individual BES-supplemented or micro-aeration-only groups (p < 0.05). This integrated approach provides a technically feasible and environmentally sustainable pathway for circular resource recovery, contributing to low-carbon agriculture and waste-to-value conversion. Full article

Plant growth-promoting rhizobacteria (PGPR) are eco-friendly and sustainable options for agrochemicals, particularly for enhancing crop productivity under stress conditions. The present research aims to isolate and characterize native PGPR from tomato rhizospheric soil and to evaluate their effectiveness as a dose-dependent response to enhance the growth of tomato seedlings. Out of 112 isolates, 10 bacterial strains were selected based on key PGPR traits, including indole-3-acetic acid (IAA), ammonia production, hydrogen cyanide (HCN), exopolysaccharide (EPS) synthesis, hydrolytic enzyme activity, potassium solubilization, antifungal activity against Fusarium oxysporum, and tolerance to pH and heat stress. Molecular identification via 16S rRNA gene sequencing confirmed that these isolates belong to the genera Serratia and Enterobacter. S. marcescens So-1 and Enterobacter sp. So-12 produced the highest levels of IAA (2.6–24.1 µg/mL). In vitro tomato seed germination tests using bacterial suspensions at three concentrations (10⁶, 10⁷, and 10⁸ CFU/mL) showed dose-dependent improvements, with T1 increasing germination up to 108.3% compared to the control. In polyhouse trials using cocopeat formulations, seedling growth improved noticeably. T2 increased the root length (28.3 ± 2.98 cm) by over 1560%, and the shoot length (35.7 ± 0.57 cm) increased by 55% against the control, whose root length is 1.7 ± 0.47. The chlorophyll amount of the treated leaves further showed significant results over the control. Collectively, these findings suggest that using native PGPR in a dose-dependent way can help tomato seedlings grow better and promote more sustainable crop production. Full article