Vítor Alves Pessoa

Non-communicable diseases (NCDs) account for more than 40 million deaths annually worldwide, including approximately 18 million premature deaths (before 70 years of age), highlighting the urgent need for effective preventive strategies. In this context, mushroom-enriched functional beverages have emerged as promising dietary approaches for the prevention and management of NCDs. This review synthesizes current evidence on mushroom-derived bioactive compounds and their application in beverage systems, integrating biochemical, technological, and regulatory perspectives. Key compounds, including β-glucans, phenolics, terpenoids, and bioactive peptides, exhibit antioxidant, anti-inflammatory, and metabolic regulatory properties, with reported effects on glycemic control, lipid metabolism, and gut microbiota modulation. Evidence from experimental and clinical studies indicates improvements such as reductions in glycated hemoglobin (HbA1c), postprandial glucose, and lipid markers, although studies specifically evaluating beverages remain limited. A patent analysis (2021–2026) identified 398 applications, of which 356 were selected, demonstrating growing technological innovation, particularly in Asia and North America. However, recent years show fewer visible applications (e.g., 60 in 2024 and 30 in 2025), likely due to confidentiality periods. Despite their functional potential, challenges related to bioavailability, clinical validation, and regulatory harmonization persist. Overall, mushroom-enriched beverages represent a promising, yet still emerging, strategy requiring further translational and clinical research to support their large-scale application in public health.

Ganoderma sp. 1962 is an Amazonian isolate with reported potential for the production of bioactive compounds; however, the effects of specific submerged fermentation parameters on mycelial architecture and on the recovery and bioactivity of protein extracts remain poorly understood. In this study, the influence of different submerged fermentation conditions on the recovery of antioxidant proteins from Ganoderma sp. 1962 was evaluated. Molecular identification was performed by sequencing the internal transcribed spacer (ITS) region followed by phylogenetic analysis. The isolate was cultivated under four nutrient compositions combined with agitated (A) and non-agitated (NA) regimes, resulting in eight experimental conditions. Mycelial biomass was structurally characterized using Scanning Electron Microscopy (SEM) and Attenuated Total Reflection–Fourier Transform Infrared (FTIR-ATR) spectroscopy. Protein extracts obtained from the mycelial biomass were analyzed for soluble protein content, total phenolic compounds (TPC), and reducing sugars (RS). Antioxidant activity was assessed using ABTS ^•+ , DPPH ^• , metal chelating ability, and reducing power assays. BLAST analysis of ITS rDNA sequences revealed high similarity (> 98% identity) to Ganoderma multiplicatum , which was subsequently confirmed by phylogenetic analysis using reference sequences from GenBank. Biomass production ranged from 5.57 to 8.73 g L ^− 1 , with marked variation in glucose consumption among conditions and a stronger tendency toward pH acidification under agitated cultivation. Macromorphological differences were predominantly characterized by pellet formation under agitated cultivation and by mycelial aggregates under static conditions. Micromorphological analysis revealed cultivation-dependent traits, including the formation of short, highly branched “staghorn” hyphae under non-agitated conditions, whereas agitated cultures promoted long, filamentous hyphae. FTIR-ATR analysis showed significant spectral differences among samples, indicating that culture medium composition strongly influenced the chemical composition of the biomass. The E3NA condition (Experiment 3, non-agitated) yielded the highest soluble protein content (218.35 ± 16.95 mg g ^− 1 extract), while E1A (Experiment 1, agitated) exhibited the highest levels of RS (25.50 ± 1.59 mg g ^− 1 extract) and TPC (31.96 ± 1.39 mg GAE g ^− 1 extract). The E1A extract showed the strongest antioxidant activity, with values of 90.34 ± 0.66% (ABTS ^•+ ), 68.09 ± 1.74% (DPPH ^• ), and 0.335 ± 0.011 (reducing power), suggesting that protein–phenol and protein–carbohydrate interactions may contribute to antioxidant activity, as supported by correlation analysis. Although non-agitated fermentation favored soluble protein recovery, protein–metabolite interactions may play a more relevant role in antioxidant activity. Overall, this study provides novel evidence of the potential of G. multiplicatum to produce antioxidant proteins under submerged fermentation.

The increasing adoption of biopolymeric and nanostructured coatings for horticultural produce has emerged as a sustainable strategy to mitigate postharvest losses and extend shelf life. However, while their technological performance has been extensively documented, comprehensive and integrative assessments of biosafety, potential human health implications, and environmental risks profiles are still insufficiently explored. This review critically analyzes recent advances in polysaccharide, protein, and lipid-based coatings, including nanoenabled systems incorporating metallic nanoparticles and bioactive agents. The mechanisms underlying gas barrier properties, antimicrobial activity, and preservation efficacy are discussed alongside degradation pathways in composting, soil, and aquatic environments. Particular attention is given to nanoparticle release, migration potential, gastrointestinal fate, and toxicological endpoints such as oxidative stress, genotoxicity, endocrine disruption, and immunomodulation. Ecotoxicological evidence across trophic levels, from microorganisms and invertebrates to fish and amphibians, is examined, highlighting sublethal and mechanistic biomarkers relevant to environmental risk assessment. Regulatory frameworks from major agencies are also compared to contextualize current safety standards and limitations. Overall, although biopolymeric coatings represent promising alternatives to conventional plastics, their life-cycle impacts, transformation products, and nano-related uncertainties require comprehensive, multilevel risk evaluation to ensure truly sustainable and safe postharvest applications.

The global functional food market continues to expand, and edible mushrooms are emerging as high-value ingredients due to their rich nutritional profile, particularly their high protein content, balanced amino acid composition, and dietary fiber. This growing industrial interest is reflected in the registration of more than 322 patents in the past five years according to the Derwent Innovation patent database. Recent advances include the integration of precision mycology (PM) and omics-based approaches, such as CRISPR-Cas9, into solid-state fermentation and submerged fermentation, enabling improvements in natural umami flavor and bioactive composition. Innovative products, including meat analogues with fibrous textures, functional beverages such as kombucha and juices, and fermented dairy products such as yogurts and cheeses, have been formulated to deliver prebiotic, antioxidant, and immunomodulatory properties. Future trends indicate a shift towards the production of high-value nutraceutical peptides and biomass, together with the adoption of artificial intelligence (AI) and the Internet of Things (IoT) to enhance bioreactor automation and scalability. Nevertheless, significant challenges remain, including regulatory constraints, the scarcity of clinical validation in humans, and the need for strict control over the bioaccumulation of heavy metals in mushroom-derived raw materials. Addressing these gaps will be critical for advancing regulatory frameworks, improving industrial standardization, and supporting the translational development of mushroom-based functional foods.

Panus lecomtei, an edible mushroom native to tropical regions, is recognized for its nutritional and bioactive compounds, particularly polysaccharides known for their immunomodulatory properties. In this study, an Amazonian strain of P. lecomtei was cultivated via submerged fermentation. The resulting mycelial biomass was subjected to intrapolysaccharide (IPS) extraction using sequential water and potassium hydroxide at 80 °C. The chemical, physicochemical, and rheological properties of the soluble fractions were analyzed using gas chromatography–mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), and dynamic light scattering (DLS). Immunostimulatory activity was assessed on macrophages by measuring cytokine release and evaluating cell adhesion to IPS-coated microbeads manipulated with optical tweezers. Fraction analysis revealed that hot-alkali extraction yielded the highest IPS recovery (0.85%) and total carbohydrate content (70%). Monosaccharide composition varied according to the extraction method. NMR analysis suggests the presence of β-glucans, α-(1→4)- and β-(1→6)-linked glucans in the hot-water fraction. The hot-alkali fraction was predominantly composed of a β-D-glucan with a (1→3)-linked main chain, substituted at O-6 by β-D-Glcp non-reducing ends or (1→6)-linked β-D-Glcp side chains. Both fractions exhibited thermal stability, low resistance to shear stress, and variable elasticity. The hot-water fraction stimulated the secretion of IL-6 and TNF-α, whereas the hot-alkali fraction induced TNF-α only. Furthermore, IPS-coated microbeads adhered to macrophage surfaces within approximately 14 s, whereas uncoated control beads required about 60 s. These findings expand current knowledge of P. lecomtei strain 1466 and support the relevance of Amazonian mushroom biodiversity as a source of bioactive macromolecules.

Edible mushrooms are an underexplored source of industrial proteases, whose synthesis is highly dependent on the cultivation substrate. This study investigated the effect of nine culture media on the proteolytic profiles of Auricularia sp., Lentinus sp., Macrocybe sp., and Grifola frondosa. Fungi were cultivated on diverse media (e.g., Czapek, Malt, Soy Flour). We analyzed total protein, specific activities (total, cysteine, serine proteases) using a biochemical assay, and protein secondary structure via FTIR, with metabolic patterns identified by PCA. A dissociation was found between total protein yield (highest in MFI/Casein media) and specific activity (highest in maltose media), suggesting catabolite repression. Distinct metabolic strategies emerged: Grifola frondosa specialized in serine protease production in the minimal Czapek medium (catabolic derepression), while Macrocybe sp. maximized cysteine protease production on soy flour (substrate induction). FTIR confirmed this, revealing a β-sheet-dominant (75.5%) structure for Grifola extract versus a random-coil-dominant (60.8%) structure for Macrocybe. This study provides a framework for mechanism-based bioprocess design, enabling the tailored production of serine proteases from G. frondosa (Czapek medium) or cysteine proteases from Macrocybe sp. (soy medium) for customized biotechnological applications.

Ganoderma sichuanense is a widely studied medicinal mushroom, but the production of its antioxidant proteins has been scarcely evaluated. We assess the influence of different concentrations of culture media components under submerged fermentation, with and without agitation, on production of proteins with antioxidant activity from the mycelial biomass of G. sichuanense. Protein extracts were characterized by scanning electron microscopy, X-ray diffraction, and attenuated total reflectance Fourier-transform infrared spectroscopy. They were also analyzed for total protein and phenolic contents, antioxidant activities (ABTS^•+, DPPH^•, chelating ability, and reducing power), and electrophoretic profiles by SDS-PAGE. The most active extract was tested for cytoprotective potential under H₂O₂-induced oxidative stress in Saccharomyces cerevisiae. Growth kinetics of the best fermentation condition were also analyzed. Microstructural differences ranged from fibrillar to aggregated forms, depending on cultivation. Crystallinity was unaffected, but chemical differences and secondary structure organization were confirmed by infrared spectroscopy. The extract from the static culture with 10 g·L⁻¹ glucose, 5 g·L⁻¹ yeast extract, and 2.5 g·L⁻¹ soy peptone (referred as CM1S) showed the highest protein and phenolic contents and the strongest antioxidant activity (IC₅₀ = 4.8 and 24.0 µg of protein·mL⁻¹ for ABTS^•+ and DPPH^•, respectively). SDS-PAGE revealed higher protein band intensities in static cultures. CM1S showed potential to protect yeast cells from oxidative stress. The Gompertz model estimated a specific growth rate of 0.0068 h⁻¹ in CM1S. The findings highlight a cultivation strategy that modulates fungal metabolism and improves the recovery of antioxidant proteins from G. sichuanense biomass.

The industrial transition to advanced biofuels is currently limited by the metabolic constraints and low inhibitor tolerance of wild-type microbial hosts. This review justifies the necessity of Precision Fermentation (PF) as the pivotal technological framework to overcome these barriers, providing a systematic synthesis of high-resolution genetic tools and intelligent bioprocess architectures. We analyze how the integration of CRISPR-Cas9, retron-mediated recombineering, and synthetic regulatory circuits enables the development of specialized microbial “chassis” capable of achieving 10- to 100-fold higher yields compared to native organisms, with industrial titers reaching 50 g/L for isobutanol and 25 g/L for farnesene. A major novelty of this work is the critical evaluation of Artificial Intelligence (AI), Soft Sensing, and Digital Twins in orchestrating real-time metabolic control and mitigating the toxic effects of advanced alcohols and drop-in hydrocarbons (C15–C20). Furthermore, the study concludes that the “scale-out” modular strategy, when integrated into hybrid thermochemical-biochemical biorefineries, allows for the full valorization of C5/C6 sugars and lignin, achieving a Minimum Selling Price (MSP) competitive with fossil fuels. By mapping the synergy between advanced metabolic engineering and data-driven process optimization, this review establishes PF as an indispensable driver for achieving carbon-neutral and carbon-negative energy systems in the circular bioeconomy.

The Amazon biome hosts a rich fungal biodiversity; however, the biotechnological potential of its macrofungi remains largely underexplored. This study evaluates and compares the mycelial growth and protease production of a wild Amazonian isolate of Pleurotus ostreatus (474) with two commercial strains (542 and 885) to identify unique biotechnological profiles. The strains were cultivated in nine different culture media, and their mycelial growth rates and major proteolytic activities were systematically measured. The wild Amazonian strain (474) exhibited significantly superior mycelial growth, reaching maximum growth on Oat Flakes Agar (OFA) within four days. In contrast, the commercial strains acted as specialized enzyme producers; strain 885 showed the highest cysteine protease activity (8.05 ± 0.15 UA/mL) on Malt Yeast Sucrose Agar (MYSA), while strain 542 exhibited the highest total protease (77.53 ± 0.67 UA/mL) and fibrinolytic activity (6.45 ± 0.00 mm²) on Malt Extract Agar (MEA). These results reveal a clear biotechnological trade-off, with the Amazonian isolate excelling in biomass accumulation and the commercial strains optimized for enzyme production. This study highlights the Amazonian strain as a promising candidate for rapid mycoprotein production and underscores the Amazon as a valuable source for expanding the functional diversity of P. ostreatus.

Background: Ganoderma spp. are a great source of bioactive molecules. The production and recovery of bioactive molecules vary according to strain, growth substrate, and extraction solution. Variations in protease and their inhibitors in basidiomata from a commercial strain (G. lingzhi) and an Amazonian isolate (Ganoderma sp.) cultivated in Amazonian lignocellulosic wastes and extracted with different solutions are plausible and were investigated in our study. Methods: Basidiomata from cultivation in substrates based on açaí seed, guaruba-cedro sawdust and three lots of marupá sawdust were submitted to extraction in water, Tris-HCl, and sodium phosphate. Protein content, proteases, and protease inhibitors were estimated through different assays. The samples were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR). Results: Tris-HCl provided higher protein extraction from Ganoderma sp. and higher caseinolytic, gelatinolytic, and fibrinolytic activity for G. lingzhi cultivated in açaí. Water extracts of Ganoderma sp., in general, exhibited higher trypsin and papain inhibitor activities compared to G. lingzhi. Extracts in Tris-HCl and sodium phosphate showed more intense protein bands in SDS-- PAGE, highlighting bands of molecular weights around 100, 50, and 30 kDa. FTIR spectra showed patterns for proteins in all extracts, with variation in transmittance according to substrate and extractor. Conclusion: Water extract from Amazonian Ganoderma sp. cultivated in marupá wastes are promising as a source of protease inhibitors, while the Tris-HCL extract of G. lingzhi from açaí cultivation stands out as a source of proteases with fibrinolytic, caseinolytic, and gelatinolytic activities.