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Since polycyclic aromatic hydrocarbons (PAHs) are mutagenic, teratogenic, and carcinogenic, they are of considerable environmental concern. A biotechnological approach to remove such compounds from polluted ecosystems could be based on the use of white-rot fungi (WRF). The potential of well-adapted indigenous Ganoderma strains to degrade PAHs remains underexplored. Seven native Ganoderma sp. strains with capacity to produce high levels of laccase enzymes and to degrade synthetic dyes were investigated for their degradation potential of PAHs. The crude enzymatic extracts produced by Ganoderma strains differentially degraded the PAHs assayed (naphthalene 34—73%, phenanthrene 9—67%, fluorene 11—64%). Ganoderma sp. UH-M was the most promising strain for the degradation of PAHs without the addition of redox mediators. The PAH oxidation performed by the extracellular enzymes produced more polar and soluble metabolites such as benzoic acid, catechol, phthalic and protocatechuic acids, allowing us to propose degradation pathways of these PAHs. This is the first study in which breakdown intermediates and degradation pathways of PAHs by a native strain of Ganoderma genus were determined. The treatment of PAHs with the biomass of this fungal strain enhanced the degradation of the three PAHs. The laccase enzymes played an important role in the degradation of these compounds; however, the role of peroxidases cannot be excluded. Ganoderma sp. UH-M is a promising candidate for the bioremediation of ecosystems polluted with PAHs.

Ganoderic acids (GAs) are bioactive triterpenoids produced by Ganoderma species with demonstrated anticancer properties. While the yield of GAs in Ganoderma tsugae is typically low, heat stress has been shown to enhance its production. This study employed atmospheric and room temperature plasma (ARTP) mutagenesis to develop thermotolerant G. tsugae mutants for high-temperature cultivation at 35°C. From 59 mutants generated, strain Ganoderma tsugae 9 (GT9) demonstrated superior thermotolerance, showing 51.48% increased mycelial growth rate and 76.03% higher biomass compared to wild-type (WT) at 35°C (one-way ANOVA with Dunnett’s test, p < 0.05). Physiological characterization revealed GT9 possessed enhanced membrane fluidity, elevated intracellular levels of lanosterol (92.49% increase), squalene (1.36-fold increase), trehalose, and ergosterol (66.98% increase) (two-way ANOVA with Tukey’s test, p < 0.05). Transcriptional analysis revealed significant upregulation of key GAs biosynthetic genes ( hmgr , sqs , se , ls ) and heat shock protein genes ( hsp17.4 , hsp22 , hsp70 , hsp90 ). After 10-day cultivation at 35°C, GT9 produced 1.01-fold more GAs than the WT at 35°C and 22.64% more than the WT at 25°C (two-way ANOVA with Tukey’s test, p < 0.05). However, the difference in GAs production between the WT strain cultured at 25°C and the GT9 strain cultured at 35°C was not significant. ARTP-generated thermotolerant G. tsugae mutants enable efficient high-temperature fermentation for enhanced GAs production. This strategy provides significant advantages for industrial-scale application while elucidating the physiological and molecular mechanisms underlying improved GAs biosynthesis under heat stress.

A white-rot basidiomycete Ganoderma spp. has long been used as a medicinal mushroom in Asia, and it has an array of pharmacological properties for immunomodulatory activity. There have been many reports about the bioactive components and their pharmacological properties. In order to analyze the current status of Ganoderma products, the detailed process of cultivation of Ganoderma spp. and development of their products are restated in this review article. These include the breeding, cultivating, extracting bioactive component, and processing Ganoderma products, etc. This article will expand people’s common knowledge on Ganoderma, and provide a beneficial reference for research and industrial production.

Fungal immunomodulatory protein (FIP) is a novel functional protein family with specific immunomodulatory activity identified from several macro-fungi. A variety of biological activities of FIPs have been reported, such as anti-allergy, anti-tumor, mitogenic activity, and immunomodulation. Among all known FIPs, the firstly discovered FIP was isolated from Ganoderma lucidum , and most FIP members were from Ganoderma genus. Compared with other FIPs, Ganoderma FIPs possess some advantageous bioactivities, like stronger anti-tumor activity. Therein, gene sequences, protein structural features, biofunctions, and recombinant expression of Ganoderma FIPs were summarized and addressed, focusing on elucidating their anti-tumor activity and molecular mechanisms. Combined with current advances, development potential and application of Ganoderma FIPs were also prospected. Key points • More than a dozen of reported FIPs are identified from Ganoderma species. • Ganoderma immunomodulatory proteins have superior anti-tumor activity with promising prospects and application. • Current review comprehensively addresses characterization, biofunctions, and anti-tumor mechanisms of Ganoderma FIPs.

Silver nanoparticles (AgNPs) are an important class of nanomaterial for a wide range of industrial and biomedical applications. AgNPs have been used as antimicrobial and disinfectant agents due their detrimental effect on target cells. The aim of our study was to determine the cytotoxic effects of biologically synthesized AgNPs using hot aqueous extracts of the mycelia of Ganoderma neo-japonicum Imazeki on MDA-MB-231 human breast cancer cells. We developed a green method for the synthesis of water-soluble AgNPs by treating silver ions with hot aqueous extract of the mycelia of G. neo-japonicum. The formation of AgNPs was characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction, dynamic light scattering, and transmission electron microscopy. Furthermore, the toxicity of synthesized AgNPs was evaluated using a series of assays: such as cell viability, lactate dehydrogenase leakage, reactive oxygen species generation, caspase 3, DNA laddering, and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling in human breast cancer cells (MDA-MB-231). The ultraviolet-visible absorption spectroscopy results showed a strong resonance centered on the surface of AgNPs at 420 nm. The X-ray diffraction analysis confirmed that the synthesized AgNPs were single-crystalline, corresponding with the result of transmission electron microscopy. Treatment of MDA-MB-231 breast cancer cells with various concentrations of AgNPs (1-10 μg/mL) for 24 hours revealed that AgNPs could inhibit cell viability and induce membrane leakage in a dose-dependent manner. Cells exposed to AgNPs showed increased reactive oxygen species and hydroxyl radical production. Furthermore, the apoptotic effects of AgNPs were confirmed by activation of caspase 3 and DNA nuclear fragmentation. The results indicate that AgNPs possess cytotoxic effects with apoptotic features and suggest that the reactive oxygen species generated by AgNPs have a significant role in apoptosis. The present findings suggest that AgNPs could contribute to the development of a suitable anticancer drug, which may lead to the development of a novel nanomedicine for the treatment of cancers.

The phytopathogenic status of G. orbiforme as the causative agent of basal stem rot on oil palm has masked its biotechnological potential. This study explored its growth profile in an Air-L-Shaped Bioreactor along with the antibacterial and antioxidant activities of its extracellular (EPS) and intracellular polysaccharides (IPS), and evaluated their toxicity using the zebrafish embryo toxicity (ZFET) assay. The biomass pellet peaked at 2.53 ± 0.44 g/L, with EPS and IPS yields reaching 0.15 ± 0.06 g/L (Day 6) and 0.06 ± 0.01 g/L (Day 10), respectively. Morphological observations revealed that the hairy starburst-shaped pellet correlated with peak EPS release. Then, the antibacterial assays revealed EPS as the better antibacterial agent compared to IPS, highlighted by a broader spectrum activity with the most prominent inhibition against Streptomyces griseus and Staphylococcus epidermidis with a minimum inhibitory concentration of 5 mg/mL. Both EPS and IPS showed strong antioxidative capabilities, demonstrated in DPPH assay—IC 50 of 15.59 ± 0.42 mg/mL for EPS and 26.85 ± 1.60 mg/mL for IPS, and FRAP assay—74.30 ± 0.38 mM Fe(II)/g for EPS and 74.18 ± 0.62 mM Fe(II)/g for IPS. Lastly, the LC 50 values of 1.88 mg/mL and 1.56 mg/mL for EPS and IPS, respectively, demonstrated their harmless nature in the ZFET assay. After 120 h of post-fertilization, the zebrafish embryos did not show abnormalities, with vital organs and structures remaining intact (fins, guts and melanophores). Collectively, this study underscores the importance of exploring the untapped potential of G. orbiforme while ensuring its safety status, opening a new possibility of bioprospecting a phytopathogen for oil palm. From a sustainable perspective, the production pattern and bioactivities are well aligned with SDG 15: Life on Land and SDG 3: Good Health and Well-Being, respectively.

Ganoderma species are major wood-decaying fungus found everywhere due to its nutritional benefits. Challenges include connecting 2D imaging of mycelium growth morphology analysis to fractal morphology and determining optimum nutrient composition to increase production yield on a larger scale in mushroom industries. In this research, a comparative study of the growth dynamics of Ganoderma lucidum and Ganoderma neo-japonicum was conducted on different nutrients including potato dextrose agar (PDA), half-strength PDA and agarose using agar plate method. Macroscopic images were used to determine average growth front length, isotropy, area and perimeter fractal dimensions, and allometric scaling exponents. The result showed that G.lucidum mycelium recorded the fastest growth rate (10.51 ± 0.018 cm on 5th day) in PDA compared to other media. Dextrose (carbon source) is necessary for mycelial growth. Meanwhile, G.neo-japonicum mycelium showed similar growth rate on PDA and half-strength PDA. G.neo-japonicum was more aggressive and could thrive even on low nutrient concentration media. Box-counting approach is used to calculate perimeter fractal dimension D and area fractal dimension D . Mycelium with weak branching has fractal dimension closer to 1.0 (sparse line-like), while dense branching has fractal dimension closer to 2.0 (area filling). The allometry scaling exponent (σ) defined as the ratio of perimeter fractal dimension to area fractal dimension is used to describe mycelium morphology. Therefore, fractal and image analysis are crucial in studying evolving complex patterns within mycelium networks. The significance of the study contributes to the large-scale mushroom industry, aligning with Sustainable Development Goal 2 (SDG2).

Ganoderma leucocontextum, a newly identified species from the Tibetan Plateau, has been mainly studied for its polysaccharides and triterpenoids, with no prior reports on fungal immunomodulatory proteins (FIPs). This study explores the biological activity of FIP-gle2, cloned from G. leucocontextum and expressed in Pichia pastoris. The effects and mechanisms of recombinant FIP-gle2 (rFIP-gle2) on cell activity and melanin synthesis in mouse melanoma B16-F10 cells were investigated in vitro. The results showed that the FIP-gle2 gene, with an open reading frame (ORF) of 333 bp, encodes a 111-amino acid polypeptide with a molecular weight of 12.60 kDa and an isoelectric point of 4.48. We achieved a yield of 184.18 mg/L of rFIP-gle2. In vitro functional experiments showed that rFIP-gle2 significantly inhibited the proliferation of B16-F10 melanoma cells and induced apoptosis in a dose-dependent manner, particularly at concentrations above 1 μg/mL. At 3 μg/mL, rFIP-gle2 effectively inhibited tyrosinase activity and reduced melanin content, downregulating microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), and tyrosinase-related proteins (TRP-1 and TRP-2). Furthermore, RNA-seq analysis indicated that differentially expressed genes in treated cells were enriched in the mitogen-activated protein kinase (MAPK) signaling pathway, with Western blotting confirming enhanced phosphorylation of JNK, ERK, and p38 proteins. Thus, P. pastoris is an effective host for rFIP-gle2 production, which shows potential for applications in pharmaceuticals, cosmeceuticals, and food fields.

Basal stem rot (BSR) of oil palm is a disastrous disease caused by a white-rot fungus Ganoderma boninense Pat. Non-ribosomal peptides (NRPs) synthesized by non-ribosomal peptide synthetases (NRPSs) are a group of secondary metabolites that act as fungal virulent factors during pathogenesis in the host. In this study, we aimed to isolate NRPS gene of G. boninense strain UPMGB001 and investigate the role of this gene during G. boninense -oil palm interaction. The isolated NRPS DNA fragment of 8322 bp was used to predict the putative peptide sequence of different domains and showed similarity with G. sinense (85%) at conserved motifs of three main NRPS domains. Phylogenetic analysis of NRPS peptide sequences demonstrated that NRPS of G. boninense belongs to the type VI siderophore family. The roots of 6-month-old oil palm seedlings were artificially inoculated for studying NRPS gene expression and disease severity in the greenhouse. The correlation between high disease severity (50%) and high expression (67-fold) of G. boninense NRPS gene at 4 months after inoculation and above indicated that this gene played a significant role in the advancement of BSR disease. Overall, these findings increase our knowledge on the gene structure of NRPS in G. boninense and its involvement in BSR pathogenesis as an effector gene.

Ganoderma lingzhi (G. lingzhi) is a Basidiomycete macrofungus valued for its secondary metabolites with pharmacological activity. To enhance the biosynthesis of secondary metabolites, various exogenous additives have been introduced to fungal fermentation processes. Metal nanomaterials, while known to influence cellular metabolism in mammalian systems, exhibit unclear effects when applied to macrofungal cultivation systems. The study systematically evaluates the impact of three distinct gold nanostructures—nanoparticles (AuNPs), nanorods (AuNRs), and nanoclusters (AuNCs)—on bioactive metabolite production during G. lingzhi submerged fermentation, employing integrated macroscopic process analytics and microscopic characterization. The results demonstrate that their impact on mycelial growth and bioactive metabolite production varied with the type, concentration, and addition timing of gold nanomaterials. Microscopic survey on cell surface morphology and nanoparticle distribution also reveals the different patterns of nanomaterial-mycelia cell interaction. Under the optimized addition conditions, AuNPs increased total polysaccharide content by 50.37% compared to the control group, while AuNRs increased triterpenoid content by 42.78%. The work confirms the potential of colloidal gold nanomaterials to facilitate the submerged fermentation of G. lingzhi , which is expected to encourage the development of nanomaterial additives-based approach for efficient microbial bioactive substances production. Graphical abstract