Antimicrobial peptides (AMPs) are promising candidates for next-generation therapeutics due to their broad-spectrum activity and reduced propensity for resistance, making them valuable in medicine, agriculture, and biotechnology. However, traditional AMP production methods including isolation from natural sources and chemical synthesis are costly, inefficient, and environmentally unsustainable, particularly for longer or post-translationally modified peptides. While heterologous expression has emerged as a scalable and versatile alternative, its success depends strongly on host selection and tailored optimisation strategies. This review examines recent advances in fungal systems as platforms for AMP production. Fungal systems, and particularly yeasts such as Pichia pastoris, offer rapid growth, low-cost fermentation, secretion capacity, and the ability to perform key post-translational modifications (PTMs), making them leading hosts for recombinant AMPs. We outline strain choice and engineering strategies that enhance AMP yield and bioactivity, including promoter and codon optimisation, secretion signal choice, fusion partners, and the construction of tandem or chimeric AMPs. By integrating current methodologies and case studies, this review aims to guide future efforts toward efficient, scalable, and commercially viable AMP manufacturing in fungal hosts, positioning fungal biotechnology as a key enabler in the development of next-generation antimicrobial solutions.

Fungi have evolved two distinct strategies, white-rot and brown-rot, to degrade lignocellulose in plant biomass. White-rot fungi utilize lignin-modifying enzymes (LMEs) to deconstruct lignin and access carbohydrates, whereas brown-rot fungi, which arose from white-rot ancestral lineages, have largely lost LME activities. Instead, brown-rot fungi rely on a small redox metabolite-mediated Fenton system that generates reactive oxygen species (ROS) to rapidly deconstruct lignocellulose and selectively remove carbohydrates. The abandonment of LMEs in brown-rot fungi suggests an intriguing evolutionary strategy to streamline decay machinery, but it remains unclear why fungi evolved this way. Here, we reintroduced LME genes from the white-rot fungus Trametes versicolor into the model brown-rot fungus Gloeophyllum trabeum to create hybrid fungal systems and evaluate LME function within the Fenton-dominant brown-rot context. Analysis of lignocellulose degradation showed that lignin disruption can be partially restored in LME mutants, thereby shifting the typical carbohydrate-selective decay mode of G. trabeum. However, constitutive LME expression also caused pronounced growth defects and reduced the overall rate of lignocellulose degradation, specifically decreasing cellulose and hemicellulose removal. Correlation analysis between genotypes and phenotypes indicates that LMEs, although prevalently used for delignification in fungi, are fundamentally incompatible with the carbohydrate-selective brown-rot system.

The opportunistic pathogenic fungus Nakaseomyces glabratus inhabits diverse host niches with fluctuating nutrient availability. Therefore, efficient control of glycolytic entry is essential, yet the regulatory principles governing hexose phosphorylation in this species remain incompletely understood. Here, we investigated the functional organization of sugar kinases in N. glabratus. Among five predicted sugar kinases, only three (Hxk2, Hxk2b and Glk1) catalyzed phosphorylation of glucose, fructose or mannose, whereas Hxk1 and Glk1b lacked detectable activity. Kinetic analyses revealed a functional specialization, with the hexokinases acting as high-capacity enzymes and Glk1 functioning as a high-affinity, low-capacity kinase optimized for low-sugar concentrations. Despite similar intrinsic kinetics among the hexokinases, Hxk2b emerged as the physiologically dominant enzyme, reflecting differential regulation rather than catalytic properties alone. Both hexokinases, but not Glk1, were strongly inhibited by trehalose-6-phosphate, linking glycolytic entry to trehalose metabolism. Consistent with this, perturbation of trehalose synthesis modulated hexose uptake, revealing that phosphorylation capacity is a major driver of sugar import, while metabolic feedback further constrains uptake. Nuclear localization of sugar kinases and condition-dependent expression patterns indicate additional regulatory layers. Together, our results demonstrate that N. glabratus controls glycolytic entry through a multilayered architecture integrating enzyme specialization, transcriptional tuning, trehalose-6-phosphate-mediated feedback and uptake coupling. This systems-level organization results in robust growth across fluctuating and often sugar-limited host environments.

Cytochalasans are a large family of fungal metabolites which inhibit actin polymerization and ultimately lead to a broad range of biological effects in different assays. Investigations into the biosynthesis of cytochalasans has revealed that the cytochrome P450 monooxygenase (P450s) tailoring enzymes possess a somewhat relaxed substrate-specificity and may accept structurally-related intermediates for oxidation, partly explaining the variety of structural variations observed in this family of molecules. In this study, we investigate a broad range of P450 enzymes via combinatorial biosynthesis to better understand their substrate scope and potential applications as biocatalysts. Genome mining enabled us to identify cryptic cytochalasan biosynthetic gene clusters (BGCs) in six different species of fungi, each with at least two P450 enzymes encoded. Comparative genomics identified a cryptic thioredoxin-like enzyme encoded in cytochalasan BGCs that co-occurs with the gene encoding a Baeyer-Villiger monooxygenase. Heterologous expression of seven P450s in Magnaporthe grisea mutant strains, lacking P450s required for pyrichalasin H biosynthesis, enabled functional characterization of three P450s, two of which were previously cryptic. The experimental results, combined with phylogenetic analysis of the P450 sequences, reveal subtle information regarding the structures of the associated cytochalasans and begins to explain why some P450s are inactive on the substrates available to them. The P450 enzymes involved in cytochalasan biosynthesis are known to be site-selective in their native host but also possess intrinsic promiscuity due to being able to modify structurally-related analogues. By investigating a diverse set of P450s from characterized and cryptic BGCs, we were able to identify that the stereochemistry of functional groups around the cytochalasan backbone is more restrictive than the size of the macrocycle when introducing the P450 enzyme to non-native substrates.

BackgroundFungi that feed and thrive on other living fungi and damage those through specific adaptations to this lifestyle are known as mycoparasites. Despite its ecological significance and practical applications in crop protection, this type of parasitism is still poorly understood. Here, we hypothesize that aggressive fungal-fungal parasitic interactions are similar to those between plants and their fungal pathogens.ResultsWe tested this hypothesis in two ways. First, we analyzed the genetic signatures of the mycoparasitic nutrition mode through the predicted Carbohydrate-Active enZYme (CAZyme) profiles of more than 50 fungi with high-quality reference genomes across the Fungal Kingdom, including mycoparasites and their close relatives. Two predicted CAZyme families, AA3-2 and AA9, appeared to be associated with mycoparasitism. Second, we searched for candidate effectors in protein datasets of three specialist mycoparasites and closely related fungi. Based on the tertiary structures of selected proteins predicted by AlphaFold, we identified protein clusters. Surprisingly, several tertiary structures predicted in three, phylogenetically diverse mycoparasites were homologous to well-studied candidate effectors in a model plant pathogen. One of these protein clusters belonged to the AA9 CAZyme family.ConclusionsThese results supported our hypothesis and may represent the first steps towards a unified molecular concept to understand mycoparasitism as a specific nutrition mode guided by candidate effectors.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40694-026-00211-5.

Glutamate dehydrogenases (GDH; EC 1.4.1.2 and EC 1.4.1.4) play a pivotal role in fungal nitrogen metabolism by catalyzing the reversible conversion of 2-ketoglutarate to L-glutamate. In fungi, NAD- as well as NADP-dependent GDHs function at the interface of ammonia assimilation and glutamate catabolism, contributing to growth, differentiation, and morphogenesis. The evolution of fungi to adapt and occupy various ecological niches is closely aligned to the diversity of regulations of the functions of GDHs, their localisation and biochemical characteristics. This review explores the biochemical, molecular, and structural studies on fungal GDHs, emphasizing their catalytic diversity, coenzyme specificity, and regulatory mechanisms, including phosphorylation, thiol modulation, and allosteric control. Structural elucidations of NADP-GDHs from Aspergillus niger, Aspergillus terreus, and Candida albicans provide new insights into cofactor binding, substrate recognition, and inhibitor interactions. Molecular analyses reveal distinct evolutionary trajectories for NAD- and NADP-GDHs across fungal taxa, with GDH-mediated transitions linked to morphogenetic processes such as the yeast-to-hypha (Y-H) switch, highlighting GDHs as promising antifungal drug targets. The comprehensive survey of fungal GDHs presented here emphasises their biochemical versatility, evolutionary significance, and translational potential in agriculture, biosensor development and in industry. The review also highlights gaps in our understanding of fungal GDHs and potential areas for further research.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40694-026-00212-4.

BackgroundFungal volatile organic compounds (FVOCs) play key roles in fungal ecology, physiology, and biotechnological applications, but inconsistent sampling and analytical methods limit biological interpretation and cross-study comparability, underscoring the need for a standardized, validated workflow.ResultsWe developed and validated a polydimethylsiloxane (PDMS)–based volatilomics workflow and evaluated its performance across key methodological dimensions, including solvent extraction bias, static versus dynamic sampling, sorbent reuse, temporal emission resolution, and discrimination of physiological states. Solvent choice (dichloromethane vs. diethyl ether) influenced the quantitative recovery of individual compounds but did not affect the overall FVOC composition detected. Static PDMS and dynamic push–pull sampling produced distinct yet complementary volatilome profiles, with method-specific enrichment across compound classes. Reconditioned PDMS tubing performed equivalently to fresh tubing across repeated deployments, with no detectable decline in compound recovery and multivariate structure. Sequential 96-h sampling captured clear temporal emission patterns in both Trichoderma atroviride and Grosmannia clavigera, revealing species-specific emission trajectories consistent with metabolic stages. Application of the optimized workflow further distinguished T. atroviride morphotypes (white vs. green), which maintained distinct volatile profiles over time and exhibited morphotype-specific emission dynamics in key compounds.ConclusionsThe PDMS-based workflow presented here provides a robust and reproducible framework for FVOC analysis, effectively addressing methodological biases, resolving temporal emission dynamics, and discriminating among physiological states. Standardizing PDMS sampling and extraction substantially enhance the biological interpretability and comparability of FVOC data, enabling broader and more reliable applications in fungal ecology, physiology, and biotechnology.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40694-026-00213-3.

BackgroundMonascus spp. are highly valuable microbial resources with extensive applications in both the food and pharmaceutical industries. In the food industry, it is often used to impart unique colors and flavors to various food products via fermentation. In the pharmaceutical field, Monascus-fermented substrate is utilized in formulating natural medicines, which exhibit beneficial properties such as lipid-lowering, antioxidant, and anti-tumor effects. However, a critical gap exists: there is currently no dedicated database for the diverse species of Monascus and its secondary metabolites. To address this, this research aims to construct a comprehensive Monascus database that meets the needs of both the research community and industry.ResultsWe successfully created the database FoodFungi (http://foodfungi.ddai.tech/). This database provides core information including: Basic details of Monascus strains; information on Monascus metabolites; relevant biological information of Monascus. Additionally, the FoodFungi database incorporates a specific function for evaluating changes in regulated Monascus products.ConclusionsThe FoodFungi database serves as a crucial support for Monascus-related research and practical applications. By providing organized, accessible information and predictive tools, it effectively promotes the further utilization of Monascus resources and drives the industrial development of Monascus-based products.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40694-026-00209-z.

Cyanodermella asteris is a fungal endophyte from Aster tataricus that produces plant hormones as well as a range of specialized metabolites. The aim of our study was to explore the potential of this endophytic fungus towards plant hormones besides the auxin indole-3-acetic acid which we recently identified. Here, we identified another hormone, jasmonic acid (JA), from culture medium extracts by LC-MS/MS and NMR. JA was also found in the hyphal fraction, but its de novo biosynthesis could not be stimulated by linolenic acid, a known precursor for JA biosynthesis in plants. The growth of C. asteris in media was not inhibited by JA. Only at high concentrations of 1 mM, an inhibition of biomass production was recorded. Putative genes encoding enzymes for JA biosynthesis were identified in the genome, and expression analyses showed an induction of one thioester hydrolase, possibly catalyzing saponification of JA-CoA to free JA. We also investigated its interaction with plant jasmonate biosynthesis and signaling mutants, aoc and jar, respectively, and found that the fungus can complement the JA-deficient phenotypes. Further understanding of the biology of JA biosynthesis on C. asteris as well as its interactions with plants is needed to exploit its potential use as a producer of JA.

Salicylic acid (SA) is an important plant hormone but is also produced by microorganisms. Contrary to the well-described roles and biosynthetic pathways of SA in plants, its role in fungal physiology and its biosynthesis within fungi remains largely unclear. Here, we sought to investigate the role of SA in the physiology of Trichoderma spp. and to identify fungal genes responsible for SA biosynthesis in Trichoderma virens, while applying and optimizing a transformation approach recently adapted for Trichoderma atroviride. Significant strain- and species-dependent differences in both SA biosynthesis and growth in the presence of exogenous SA were observed. Furthermore, in certain Trichoderma species SA biosynthesis turned out to be induced by the presence of plant volatile organic compounds (VOCs). Based on plant SA biosynthesis pathways, candidate fungal SA biosynthesis genes were screened and respective T. virens gene deletion mutants generated through application and optimization of an enhanced transformation approach. Gene deletion did not result in a decrease in SA biosynthesis, providing evidence that SA biosynthesis in T. virens is distinct from the canonical plant pathways. Although we were not able to identify genes responsible for SA biosynthesis in T. virens, we uncovered how certain Trichoderma and fungal phytopathogen species are affected by SA in their environment and how SA release by Trichoderma spp. can be affected by the presence of a plant host. Furthermore, we were able to optimize an approach to measuring phytohormones produced by Trichoderma spp. in plate culture and proved the applicability of an optimized transformation approach in T. virens.