Search Results (69)

Fungal hydrophobins reduce the surface tension of hyphae so that hyphae can grow into the air. Reduced expression of hydrophobin genes results in abnormal morphogenesis of both hyphae and the fruiting body of Flammulina filiformis. Previous studies showed that filamentous-growth MAPK signaling pathway directly modulates pseudohyphae formation in budding yeast, so we hypothesized that the specific transcription factor in this pathway may also directly regulate the expression of hydrophobin genes in F. filiformis. Downstream of the G protein, the cAMP/PKA signaling pathway is parallel with the filamentous-growth MAPK signaling pathway in regulating the filamentous growth of fungi. Thus, the cAMP addition test was carried out to exclude the involvement of the PKA/cAMP signaling pathway in aerial-hyphae deficiency of the three mutants used in our previous study. Transcriptomic analysis showed common changes in the MAPK signaling pathway of the three mutants, including 6 downregulated and 3 upregulated genes in common. Transcription factor Tec1 was one of the upregulated genes, and it is a pathway-specific transcription factor for filamentous growth. Motif prediction showed that putative binding sites of Tec1 and Ste12 existed in the promoter region of the three chosen hydrophobin genes mentioned in our previous study, and DAP-seq analysis suggested that putative binding sites of Tec1 and Ste12 were located in 10 hydrophobin genes, respectively, and there were 8 in common for both the transcription factors. These results gave suggestive evidence supporting our hypothesis. We have identified a potential regulatory connection between the filamentous-growth MAPK signaling pathway and hydrophobin genes through Tec1 and Ste12. However, functional validation is required to confirm direct regulation between both the transcription factors and the downstream genes. Full article

Surface fouling remains a critical challenge for medical devices and chemosensor systems operating in biological environments, where nonspecific adsorption of proteins, cells, and microorganisms can lead to signal drift, reduced sensitivity, and shortened device lifetime. Conventional antifouling strategies rely primarily on synthetic hydrophilic polymer coatings, such as polyethylene glycol and polyvinylpyrrolidone, which are effective but face limitations related to long-term stability, thickness, and compatibility with surface-sensitive sensing modalities. In this review, we focus on hydrophobins derived from mushroom-forming and filamentous fungi as a bio-based alternative for antifouling and anti-wetting surface modification. Mushroom-derived hydrophobins are small amphiphilic proteins capable of spontaneous self-assembly into nanometer-scale films that modulate surface energy, wettability, and interfacial friction without requiring covalent functionalization. The current state of research on hydrophobin structure, classification, and self-assembly is reviewed, followed by a synthesis of reported antifouling and tribological behaviors relevant to medical and sensor-adjacent surfaces. Representative experimental observations are discussed to illustrate trends consistent with the literature, without establishing new performance benchmarks. The implications of mushroom-derived hydrophobin coatings for chemosensors and biosensors are examined, particularly with respect to signal stability, surface accessibility, and durability. Limitations and future research directions are outlined to support translation into practical sensing technologies. Full article

Boletus griseipurpureus is an ectomycorrhizal mushroom characterized by a bitter flavor. In this study, specimens were collected from three host plants—Acacia auriculiformis (BgAa), Melaleuca cajuputi (BgMc), and Eucalyptus camaldulensis (BgEc)—and initially classified based on pileus morphology. Molecular biodiversity was investigated using internal transcribed spacer (ITS) DNA barcoding, and comprehensive phylogenetic analysis revealed that B. griseipurpureus populations in southern Thailand clustered according to their symbiotic host species. De novo transcriptome assembly of B. griseipurpureus associated with different hosts was performed to generate unigene datasets, followed by functional gene annotation. A total of 1157 differentially expressed genes (DEGs) were identified and linked to ectomycorrhizal symbiosis. The genes involved in biosynthesis and metabolic processes exhibited host-dependent expression patterns. Furthermore, expression profiles of five selected genes—major facilitator superfamily (MFS) substrate transporter, phosphatase II, hexose transporter, terpenoid synthase, and fungal hydrophobin—were consistent between DEG analysis and semi-quantitative RT-PCR validation. These findings suggest that these genes play important roles in ectomycorrhizal symbiosis and the biosynthesis of bioactive compounds in B. griseipurpureus, with expression influenced by host association. This study provides valuable insights into the molecular biodiversity and gene regulation underlying ectomycorrhizal symbiosis, contributing to a better understanding of the biological processes in B. griseipurpureus. Full article

PET biodegradation remains limited due to its intrinsic properties—high crystallinity, hydrophobicity, and strong chemical stability. These characteristics lead to extremely slow degradation rates and contribute to PET’s persistence in the environment. Understanding how microorganisms respond at the molecular level when exposed to such a recalcitrant polymer is therefore essential. Living organisms express genes in response to their needs during development. When microbes are under critical conditions, such as when contaminants are present, they express genes encoding specific enzymes that attack the pollutant. In this study, a fungus isolated from the infected fruit of the plant Randia monantha was identified as Aspergillus terreus. It was tested for polyethylene terephthalate (PET) degradation, and the fungus Aspergillus nidulans was evaluated due to its previously reported recombinant cutinases for PET degradation. A microplastic polyethylene terephthalate (PET-MP) particle size of <355 μm for degradation was established, and a PET weight loss of 1.62% for A. nidulans and 1.01% for A. terreus was found. Additionally, the degradation of PET was confirmed by FTIR and SEM. This study also compares the transcriptomic profiles of Aspergillus nidulans and Aspergillus terreus during cultivation with PET-MP residues, which serve as a replacement for the carbon source. We present the first evidence of chitinase overexpression during direct exposure of PET to Aspergillus fungi. Interestingly, chitinase expression was detected in the crude extracts of A. nidulans and A. terreus during culture in the presence of PET residues, which replaced the carbon source. The chitinase produced by each fungus has a similar molecular weight of approximately 44 kDa. Chitinase activity was monitored over a 14-day cultivation period; from day 2, chitinase activity was detected in both cultures and continued to increase until day 14, when the highest values reported in this work were 24.88 ± 4.17 U mg⁻¹ and 10.41 ± 0.47 U mg⁻¹ for A. nidulans and A. terreus, respectively. Finally, we proposed a pathway for PET degradation by Aspergillus fungi that involves mycelial adherence and the secretion of hydrophobins, followed by the production of intermediates and monomers via esterase hydrolysis, and ultimately, the entry of monomers to the ethylene glycol (EG) and terephthalic acid (TPA) pathways, further suggesting these Aspergillus as candidates to produce valuable compounds under these conditions, such as muconic acid, gallic acid, and vanillic acid. Full article

Ectomycorrhizal (ECM) symbiosis is a fundamental mutualism crucial for forest eco-system health. Its establishment is governed by sophisticated molecular dialogue preceding physical colonization. This review synthesizes this pre-colonization crosstalk, beginning with reciprocal signal exchange where root exudates trigger fungal growth, and fungal lipochitooligosaccharides activate host symbiotic programming, often via the common symbiosis pathway. Successful colonization requires fungi to navigate plant immunity. They employ effectors, notably mycorrhiza-induced small secreted proteins (MiSSPs), to suppress defenses, e.g., by stabilizing jasmonate signaling repressors or inhibiting apoplastic proteases, establishing a localized “mycorrhiza-induced resistance.” Concurrent structural adaptations, including fungal hydrophobins, expansins, and cell wall-modifying enzymes like chitin deacetylase, facilitate adhesion and apoplastic penetration. While this sequential model integrates immune suppression with structural remodeling, current understanding is predominantly derived from a limited set of model systems. Significant knowledge gaps persist regarding species-specific determinants in non-model fungi and hosts, the influence of environmental variability and microbiome interactions, and methodological challenges in capturing early signaling in situ. This review’s main contributions are: providing a synthesized sequential model of molecular crosstalk; elucidating the dual fungal strategy of simultaneous immune suppression and structural remodeling; and identifying crucial knowledge gaps regarding non-model systems and species-specific determinants, establishing a research roadmap with implications for forest management and ecosystem sustainability. Full article

Sclerotinia sclerotiorum is a notorious soilborne fungal pathogen that causes white mold in a wide range of host plants, leading to globally significant yield loss in many crops. Hydrophobins (HPs) are small, secreted proteins unique to filamentous fungi, with diverse roles in fungal biology. However, their functions in S. sclerotiorum remain poorly understood. Here, we systematically investigated the roles of three HP genes, SsHP1, SsHP2, and SsHP3, through reverse genetic analyses. By analyzing their deletion mutant phenotypes, we demonstrate that class I HP (SsHP1) is specifically required for proper sclerotia development, whereas class II HPs (SsHP2 and SsHP3) are essential for compound appressoria functionality. All three HPs contribute to fungal surface hydrophobicity, cell wall integrity, and stress tolerance. Using mycelial fusion, we generated double mutants lacking both class II HPs, which exhibited more severe defects in appressoria development, virulence, cell wall integrity, and stress adaptation, indicating their partially redundant roles. SsHP2 is required for both host penetration and post-penetration virulence, whereas SsHP3 mainly affects host penetration, revealing their overlapping yet distinct contributions to pathogenic development. Although all HP mutants formed normal apothecia and asci, they released significantly fewer ascospores, suggesting that HPs are dispensable for sexual morphogenesis but crucial for the biophysical process of ascospore dispersal. Furthermore, carbohydrate analyses uncovered that these HPs affect cell wall composition, more broadly influencing stress adaptation and virulence. Taken together, our study reveals both conserved and divergent roles of HPs across fungi and highlight their multifaceted contributions to S. sclerotiorum biology, offering new perspectives for disease management. Full article

Sarcomyxa edulis is a characteristic edible and medicinal mushroom found in Northeast China that is highly valued by consumers for its tender texture, pleasant flavor, and high nutritional value. To gain a deeper understanding of the molecular mechanisms underlying the development of S. edulis fruiting bodies, this study utilized the Illumina NovaSeq platform to perform transcriptome sequencing at three growth and development stages of S. edulis strain SE8, namely primordia (SE8–P), fruiting body differentiation (SE8–F), and mature fruiting body (SE8–M). A total of 54.67 Gb of clean data was obtained, with a GC content of around 51%. After assembly, 36,423 Unigenes were obtained. Functional annotation was performed on the Unigenes, resulting in 21,206 Unigene annotation results. Differential expression gene analysis showed that 79,606 and 523 DEGs were annotated in at least one database during the SE8–P vs. SE8–F, SE8–F vs. SE8–M, and SE8–P vs. SE8–M processes, respectively. Among these, the genes encoding aldehyde dehydrogenase and fungal hydrophobins were consistently downregulated, playing a negative regulatory role in the growth and development of S. edulis. The genes encoding glycoside hydrolase and AB hydrolase superfamily proteins were consistently upregulated, playing a positive regulatory role in growth and development. Among these, the genes encoding aldehyde dehydrogenase were annotated to the Tryptophan metabolism (ko00380) pathway through KEGG, suggesting that aldehyde dehydrogenase regulates indoacetate formation in the fruiting body of S. edulis. The accuracy of RNA–Seq and DEG analysis was validated using quantitative PCR. This study enriches our knowledge of the genetic information and provides a theoretical basis for the molecular mechanisms of fruiting body development of S. edulis. Full article

The global transition to a circular bioeconomy is accelerating the demand for sustainable, high-performance materials. Filamentous fungi represent a promising solution, as they function as living foundries that transform low-value biomass into advanced, self-assembling materials. While mycelium-based composites have proven potential, progress has been predominantly driven by empirical screening of fungal species and substrates. To unlock their full potential, a paradigm shift from empirical screening to rational design is required. This review introduces a conceptual framework centered on the biochemical programming of the fungal cell wall. Viewed through a materials science lens, the cell wall is a dynamic, hierarchical nanocomposite whose properties can be deliberately tuned. We analyze the contributions of its principal components—the chitin–glucan structural scaffold, the glycoprotein functional matrix, and surface-active hydrophobins—to the bulk characteristics of mycelium-derived materials. We then identify biochemical levers for controlling these properties. External factors such as substrate composition and environmental cues (e.g., pH) modulate cell wall architecture through conserved signaling pathways. Complementing these, an internal synthetic biology toolkit enables direct genetic and chemical intervention. Strategies include targeted engineering of biosynthetic and regulatory genes (e.g., CHS, AGS, GCN5), chemical genetics to dynamically adjust synthesis during growth, and modification of surface chemistry for specialized applications like tissue engineering. By integrating fungal cell wall biochemistry, materials science, and synthetic biology, this framework moves the field from incidental discovery toward the intentional creation of smart, functional, and sustainable mycelium-based materials—aligning material innovation with the imperatives of the circular bioeconomy. Full article

Edible mushrooms represent great promise for the future of food and medicine due to their excellent nutritional, functional, and therapeutic properties. Macrofungi synthesize numerous bioactive compounds, among which proteins stand out for their remarkable diversity, both in terms of structure and their nutritional and functional roles. Fungi from the phylum Basidiomycota have a high protein content, characterized by a complete and balanced amino acid composition. Proteins and peptides from mushrooms have both nutritional and functional roles, with numerous health benefits, such as antimicrobial, antiviral, antioxidant, anticancer, hypotensive, angiotensin-converting enzyme (ACE) inhibition, immunomodulatory, and enzymatic activities. Functional proteins include lectins, immunomodulatory proteins, enzymes (laccase, cellulase, ribonuclease), enzyme inhibitors, ribosome-inactivating proteins, and hydrophobins. In addition to traditional cultivation, mushrooms can be grown as mycelium on solid substrates or in submerged culture, followed by protein separation and extraction. The main trends in protein biosynthesis from Basidiomycota involve both improving the properties of the producing strains and optimizing the cultivation methods in submerged culture and on solid substrates. Moreover, new techniques in the fields of genomics, proteomics, and metabolomics will enable increasingly promising results. This paper provides a systematic overview of the types and properties of proteins from edible mushrooms, with a focus on the main beneficial effects of their consumption. Full article

Hydrophobins are small, surface-active protein biosurfactants secreted by filamentous fungi with potential applications in industries such as pharmaceuticals, sanitation, and biomaterials. Additionally, hydrophobins are known to stabilize enzymatic processing of biomass for improved catalytic efficiency. In this study, Pichia pastoris was used to recombinantly express hydrophobin HFBI from Trichoderma reesei, a well-characterized fungal system used industrially for bioethanol production. Iterative optimization was performed on both the induction and purification of HFBI, ultimately producing yields of 86.6 mg/L HFBI and elution concentrations of 48 μM HFBI determined pure by SDS-PAGE, over a five-day methanol-fed batch shake flask induction regiment followed by a single unit operation multimodal cation exchange purification. This final purified material represents an improvement over prior approaches to enable a wider range of potential applications for biosurfactants. Full article

Protoplasts are essential tools for genetic manipulation and functional genomics research in fungi. This study systematically optimized protoplast preparation conditions and examined transcriptional changes throughout the preparation and regeneration processes to elucidate the molecular mechanisms underlying the formation and regeneration of protoplasts in Lyophyllum decastes. The results indicated an optimal protoplast yield of 5.475 × 10⁶ cells/mL under conditions of fungal age at 10 days, digestion time of 2.25 h, enzyme concentration of 2%, and digestion temperature of 28 °C. The Z5 medium supplemented with L. decastes mycelial extract achieved a high regeneration rate of 2.86. RNA-seq analysis revealed 2432 differentially expressed genes (DEGs) during protoplast formation and 5825 DEGs during regeneration. Casein kinase I, cytochrome P450 (CYP52), and redox-regulated input receptor (PEX5) were significantly upregulated during the protoplast stage, while β-1,3-glucan synthase (SKN1), chitin synthase (CHS2), hydrophobin-1, and hydrophobin-2 showed significant upregulation during the protoplast regeneration phase. These findings provide a reference for the efficient preparation and regeneration of protoplasts and offer new insights into the molecular mechanisms of protoplast formation and cell wall regeneration in fungi. Full article

Background: The properties of nanoparticle surfaces are crucial in influencing their interaction with biological environments, as well as their stability, biocompatibility, targeting abilities, and cellular uptake. Hydrophobin 4 (HFB4) is a class II HFB protein produced by filamentous fungi that has a natural ability to self-assemble at hydrophobic-hydrophilic interfaces. The biocompatible, non-toxic, biodegradable, and amphipathic properties of HFB4 render it valuable for improving the solubility and bioavailability of hydrophobic drugs. We have investigated the physicochemical properties, cellular uptake, and anticancer effects of empty and Doxorubicin (Dox)-loaded HFB4 liposomes (HFB4L) and compared them to those of PEGylated liposomes (PPL). Methods: The Pichia pastoris KM71H strain was used for HFB4 purification. Liposomes were prepared through the thin film hydration method and characterized. The cytotoxic effects of free Dox, Dox-HFB4, and Dox-PPL were assessed in MCF7 cells using the SRB Assay. Results: All formulations showed good size homogeneity and a spherical shape. The HFB4 coating enhanced the physicochemical stability of Dox-HFB4L over 60 days at 4 °C without significantly affecting Dox release from HFB4L. It increased Dox release at pH 5.4 compared to pH 7.4, indicating higher delivery of drugs into acidic tumor environments, similar to Dox-PPL. While both formulations showed increased cellular uptake compared to free Dox, they exhibited a lower anticancer effect due to the sustained release of Dox. Notably, Dox-HFB4L displayed greater cytotoxicity than Dox-PPL in MCF7 cells. Conclusions: HFB4L may offer superior benefits in terms of delivering drugs to an acidic tumor environment in a stable, non-toxic, and sustained manner. Full article

Fungal diseases not only reduce the yield of edible mushrooms but also pose potential threats to the preservation and quality of harvested mushrooms. Cobweb disease, caused primarily by fungal pathogens from the Hypocreaceae family, is one of the most significant diseases affecting edible mushrooms. Deciphering the genomes of these pathogens will help unravel the molecular basis of their evolution and identify genes responsible for pathogenicity. Here, we present high-quality genome sequences of three cobweb disease fungi: Hypomyces aurantius Cb-Fv, Cladobotryum mycophilum CB-Ab, and Cladobotryum protrusum CB-Mi, isolated from Flammulina velutipes, Agaricus bisporus, and Morchella importuna, respectively. The assembled genomes of H. aurantius, C. mycophilum, and C. protrusum are 33.19 Mb, 39.83 Mb, and 38.10 Mb, respectively. This is the first report of the genome of H. aurantius. Phylogenetic analysis revealed that cobweb disease pathogens are closely related and diverged approximately 17.51 million years ago. CAZymes (mainly chitinases, glucan endo-1,3-beta-glucosidases, and secondary metabolite synthases), proteases, KP3 killer proteins, lipases, and hydrophobins were found to be conserved and strongly associated with pathogenicity, virulence, and adaptation in the three cobweb pathogens. This study provides insights into the genome structure, genome organization, and pathogenicity of these three cobweb disease fungi, which will be a valuable resource for comparative genomics studies of cobweb pathogens and will help control this disease, thereby enhancing mushroom quality. Full article

Hydrophobins are proteins, consisting of approximately 70–130 amino acids and containing eight cysteines, linked by four disulfide bonds, which are characteristic of the entire hydrophobin family. The main advantage of hydrophobins is their ability to form amphiphatic layers on surfaces and thus to change their properties from hydrophilic to hydrophobic and vice versa. It is for this reason that hydrophobins can be widely used in a variety of applications to improve the properties of materials, such as hydrophilicity, activity and stability of immobilized molecules. In this work, the hydrophobin RodA of Aspergillus fumigatus and its properties were investigated. The gene responsible for the synthesis of the RodA protein was identified by molecular biology methods and used to design an expression system. The purified recombinant RodA protein was used to modify the surface of a gold electrode in order to investigate the effect of this hydrophobin as a matrix on the performance of the engineered glucose biosensor. The engineered biosensor with the RodA matrix was compared with a biosensor without the RodA matrix. The data obtained were fitted to Michaelis–Menten and linear models to calculate the K_M and the maximum current generated (I_max). In the case of Au/GOx, the K_M value was 6.99 mM and the I_max was 34.8 μA·cm⁻²; in the case of the Au/RodA/GOx biosensor, the K_M value was 2.37 mM and the I_max was 0.432 μA·cm⁻². The lower I_max value for the Au/RodA/GOx biosensor could be explained by the possible formation of an excessively thick monolayer of RodA protein or by possible conformations of the protein that blocked the glucose oxidase molecules. However, the K_M value obtained for Au/RodA/GOx showed that for this biosensor, the immobilized glucose oxidase has a significantly higher affinity for the substrate, indicating that such a protein may be suitable for electrode modifications. Full article

Gas-filled microbubbles are well-established contrast agents for ultrasound imaging and widely studied as delivery systems for theranostics. Herein, we have demonstrated the promising potential of the hydrophobin HFBII—a fungal amphiphilic protein—in stabilizing microbubbles with various fluorinated core gases. A thorough screening of several experimental parameters was performed to find the optimized conditions regarding the preparation technique, type of core gas, HFBII initial concentration, and protein dissolution procedure. The best results were obtained by combining perfluorobutane (C₄F₁₀) gas with 1 mg/mL of aqueous HFBII, which afforded a total bubble concentration higher than 10⁹ bubbles/mL, with long-term stability in solution (at least 3 h). Acoustic characterization of such microbubbles in the typical ultrasound frequency range used for diagnostic imaging showed the lower pressure resistance of HFBII microbubbles, if compared to conventional ones stabilized by phospholipid shells, but, at the same time, revealed strong non-linear behavior, with a significant harmonic response already at low acoustic pressures. These findings suggest the possibility of further improving the performance of HFBII-coated perfluorinated gas microbubbles, for instance by mixing the protein with other stabilizing agents, e.g., phospholipids, in order to tune the viscoelastic properties of the outer shell. Full article