Extracellular vesicles (EVs) from the edible mushroom Pleurotus tuber-regium (PTR) were investigated with respect to their environmental responsiveness, molecular features, and preliminary functional properties. PTR-EVs were characterized by dynamic light scattering, nanoparticle tracking analysis, and transmission electron microscopy. Proteomic analysis revealed enrichment of ribosomal and proteasomal proteins, redox-related enzymes, and vesicle trafficking components, suggesting non-random molecular representation. Small RNA sequencing identified abundant novel miRNAs with predicted targets involved in nitrogen metabolism, cell wall remodeling, redox regulation, and ubiquitin-mediated proteolysis. Among the tested factors, temperature showed the strongest association with vesicle production, with particle concentration increasing from 1.22 × 109 to 7.31 × 109 particles/mL at 34 °C, approximately six-fold higher than at 30 °C. Transcriptomic profiling showed coordinated repression of cell wall-associated genes and redox enzymes, together with induction of endoplasmic reticulum proteostasis pathways, consistent with stress-associated changes in the cellular context of vesicle release. Ultrasonicated PTR-EVs exhibited enhanced DPPH and ABTS radical-scavenging activities in chemical assays, with DPPH increasing from 59.52% to 71.73% and ABTS from 38.25% to 40.51%. Encapsulation efficiencies reached 32.67% ± 1.3% for proanthocyanidins and 46.01% ± 0.5% for curcumin. PTR-EVs showed the best short-term stability at pH 7 and 4 °C, supporting their further evaluation as an edible fungal vesicle platform for food-related nanoscale delivery.

Two β-1,3-glucosyltransferase isoenzymes, designated GL20535 and GL24465, are present in Ganoderma lucidum polysaccharide (GLPS) biosynthesis. Previous studies have demonstrated that the expression level of gl20535 significantly influences the yield of GLPS, and silencing gl20535 results in upregulation of gl24465. However, the specific function of GL24465 in polysaccharide biosynthesis and its interaction mechanism with GL20535 remain poorly understood. In this study, the roles and interrelationship between GL24465 and GL20535 in GLPS biosynthesis were systematically investigated through individual and combinatorial gene silencing and overexpression approaches. Results indicated that individual modulation of gl24465 exerted minimal effects on polysaccharide production. In contrast, co-overexpression of both genes markedly enhanced the accumulation of intracellular polysaccharides (IPS) and extracellular polysaccharides (EPS), with increases of 9.92% and 89.15%. Conversely, co-silencing led to significant reductions in cell biomass, cell wall thickness, and EPS and IPS contents, by 13.81%, 50.25%, 20.52%, and 35.02%, respectively. Co-regulation also affected GLPS monosaccharide composition and glycosidic linkage ratios by altering the transcription of key enzymes in sugar donor synthesis, including phosphoglucomutase (PGM) and UDP-glucose pyrophosphorylase (UGP), as well as glucosyltransferases Transcriptional analysis revealed that changes in gl24465 expression did not significantly affect the transcription of gl20535. Integrating these findings with existing evidence indicating that GL20535 functions as a major enzyme in polysaccharide production, while GL24465 provides compensatory support for maintaining both yield and structural integrity, this study offers novel insights into the functional cooperation of glycosyltransferase isoenzymes in edible and medicinal fungi，with potential implications in biomanufacturing optimized polysaccharides.

Glycosylation is a common modification in natural products that enhances their pharmacological value. Icariside D2, derived from the O-glycosylation of tyrosol, exerts significant antitumor and cardiovascular protective effects; however, biosynthesis is limited by poor UDP-glycosyltransferase (UGT) regioselectivity, which results in mixtures of icariside D2 and salidroside. By targeting the glycosyltransferase from Bacillus licheniformis ZSP01 (UGTBL1), we pinpointed a tunnel near the nucleophilic histidine in the active pocket (tunnel X) that was not essential for substrate entry or product release, yet it efficiently controlled regioselectivity. Blocking tunnel X resulted in exclusive regioselectivity toward icariside D2 synthesis, with enzyme activity 1.8-fold that of the wild type, overcoming the tradeoff between regioselectivity and enzyme activity. Regioselectivity and structural analysis revealed that blocking tunnel X hampered deprotonation of the alcohol hydroxyl in tyrosol, thereby hindering salidroside synthesis. Notably, glucosylation of the phenolic hydroxyl group remained favored for icariside D2 synthesis. The proposed tunnel-blocking strategy was applied to another UGT from Bacillus subtilis 168 (BSYjiC), which produced BS-A13E and BS-A13H mutants with elevated regioselectivity (77.2% and 78.7%, respectively) toward the phenolic hydroxyl of tyrosol. This study provides a theoretical basis for the exclusive and efficient biosynthesis of icariside D2 and offers insights into the regioselective modification of UGTs.

β-1,3-Glucosyltransferase is a key glycosyltransferase that affects the biosynthesis of polysaccharides in Ganoderma lucidum, an edible and medicinal fungal polysaccharide. Its isoenzymes, GL20535 and GL24465, serve dominant and auxiliary roles, respectively. Although the catalytic activity of β-1,3-glucosyltransferase is regulated by Rho GTPases, the mechanisms by which these regulators affect GL20535 and GL24465 and consequently influence polysaccharide synthesis remain unclear. In the present study, three members of the G. lucidum Rho GTPase (GLRho) family—GLRho1/GLRho2/GLRho3—were identified by gene mining. Through AlphaFold3 modeling and protein–protein interaction analysis, it was found that GLRho1 and GLRho3 possess similar structures, binding primarily to GL20535. However, GLRho exhibited stronger binding capacity to GL24465 due to differences in secondary structure by computational predictions. Polysaccharide metabolism and transcription analysis showed that GLRho1 and GLRho3 exerted positive effects on growth and polysaccharide synthesis in G. lucidum, with the overexpression of both genes upregulating the expression of gl20535, leading to significant increases of 27.65

Background Numerous species of filamentous fungi are sustainable and potent hosts for food biotechnology, owing to their intimate association with the development and utilization of food resources. Unlike bacteria and yeasts, filamentous fungi exhibit a remarkable level of morphological complexity, endowing them with unique properties particularly advantageous for food industrial applications. Scope and approach This review summarizes recent advances in filamentous fungi, with a focus on mycelial morphology in submerged culture and its implications for food biomanufacturing. It addresses (i) morphological characteristics; (ii) the impact of morphology on process performance; (iii) morphological shaping strategies—ranging from physical, genetic to microbial interaction; and (iv) advanced monitoring techniques. Furthermore, the study proposes an upgrading fungal cell factory enabled by an integrated morphology engineering platform that combines genetic engineering, synthetic biology, metabolic engineering, and fermentation process optimization. Challenges associated with the application of engineered mycelia in food biomanufacturing are also examined. Findings and conclusion Filamentous fungi process advantage of remarkable morphological plasticity and metabolic diversity, and precise control of mycelial morphology is essential for unlocking their industrial potential. The integration and advancements in morphology engineering and morphology metabolism engineering—encompassing morphological shaping, monitoring, and editing—enable precise morphology control. The resulting capacity to customize morphology is essential for engineering mycelium as innovative food ingredients and high-performance biomaterials. Through such collaborative innovation, morphological engineering is poised to transform filamentous fungi into robust, efficient, and sustainable cell factories, offering novel solutions to global food sustainability challenges.

Mycelium-based materials represent a promising sustainable alternative to petroleum-based plastics. However, their broad-scale application remains hindered by intrinsic mechanical frailty and the ubiquitous strength-toughness trade-off. In this work, we present a bottom-up fabrication strategy that overcomes these limitations by engineering desirable material properties during the biological growth phase. Specifically, by introducing Tween-20 as a bio-regulator during the liquid culture of Ganoderma lucidum, thereby enabling programmable, synergistic multiscale structural reinforcement through endogenous fungal metabolism. At the macroscale, this approach orchestrates the spontaneous self-assembly of a densely entangled three-dimensional hyphal network, significantly enhancing load transfer. At the microscale, Tween-20 modulates cell wall composition and fatty acid biosynthesis, priming the matrix for robust covalent ester cross-linking with a glycerol-based plasticizer. Critically, Tween-20 functions as a metabolic elicitor that directs in situ structural programming and chemical consolidation within a single fabrication cycle. The resulting biomaterial achieves outstanding mechanical performance, with a tensile strength of 24.21 MPa, a Young's modulus of 116.45 MPa, and a toughness of 2909.47 kJ m-3, substantially surpassing the corresponding values obtained for previously reported pure mycelium-based materials. Moreover, the material demonstrates integrated multifunctionality, including hydrophobicity, thermal stability, antibacterial activity, and biodegradability. This study establishes a scalable, environmentally benign platform for high-performance biocomposites manufacturing and provides a foundational design paradigm for next-generation sustainable consumer goods and packaging solutions.

Enzyme immobilization is crucial for industrial biocatalysis but is fundamentally limited by an inherent activity-stability trade-off, where rigid protective supports often compromise catalytic efficiency through severe diffusion barriers or denaturation. To address this challenge, we developed a bioinspired "nanoarmor" via interfacial assembly on polyethylenimine-functionalized styrene maleic anhydride nanospheres (PEI-SMA). Initially, crude sucrose synthase was immobilized through electrostatic adsorption onto cationic PEI-SMA surfaces, effectively bypassing purification and mitigating denaturation caused by subsequent precursor ions. A synergistic PEI-sodium alginate (SA) system then enabled protective coating formation: PEI pre-configured the reaction sites by coordinating Zn2 + at the nanosphere interface, while SA induced instantaneous nucleation via biomimetic coordination with substoichiometric 2-methylimidazole (2-MeIm). This dual-control strategy, combining non-equilibrium ligand ratios with rapid nucleation kinetics, suppressed crystal growth and directed conformal amorphous ZIF deposition (PEI-SMA@enzyme-Sazif). This core-shell colloidal architecture circumvents diffusion limitations via its amorphous matrix while ensuring superior stabilization. The biocatalyst exhibited high activity retention (71.4%) and favorable reusability (75.7% after 5 cycles), outperforming PEI-SMA@enzyme. Furthermore, it showed significantly enhanced environmental stability, exhibiting 2.6-fold and 1.3-fold higher residual activity than the free enzyme after incubation at 60 °C and pH 5.5 for 60 min, respectively. This work illustrates how rational design of colloidal biointerfaces and interfacial biomineralization pathways can reconcile the stability-activity dilemma, enabling efficient and stable biocatalysis directly from crude enzyme extracts.

Sulfated derivatives of hyperbranched beta-glucans from the edible mushroom Pleurotus tuber-regium (S-HBGs) were synthesized with varying degrees of substitution (DS) in the current study to evaluate their hypoglycemic and anti-glycation efficacy. Characterization analysis revealed that sulfation minimally affected the polymer's branched structure but reduced its molecular weight. Transmission electron microscopy (TEM) confirmed that the branched and interwoven chain conformation of S-HBGs remained largely intact despite the presence of sulfate groups. S-HBGs demonstrated reversible inhibition of alpha-glucosidase, with the inhibition mode transitioning from competitive to mixed-type and eventually to non-competitive as DS decreased from >0.70 to 0.55 and 0.27. Furthermore, S-HBGs with moderate or low DS effectively inhibited protein glycation and the formation of advanced glycation end products (AGEs). These findings advance the understanding of S-HBGs as novel hypoglycemic and anti-glycation agents, underscoring their potential as bioactive food components for managing diabetes and its complications.

Ganoderma lucidum polysaccharides (GLPs) are natural compounds with a broad spectrum of biological activities. β-1,3-glucosyltransferase (GL20535) plays an important role in polysaccharide synthesis by catalyzing the transfer of UDP-glucose to extend sugar chains, but its underlying mechanism remains unclear. In this study, the regulatory mechanism of GL20535 in polysaccharide synthesis was elucidated by overexpressing and silencing gl20535 in G. lucidum. Overexpression of gl20535 resulted in maximum increases of 18.08%, 79.04%, and 18.01% in intracellular polysaccharide (IPS), extracellular polysaccharide (EPS), and β-1,3-glucan contents, respectively. In contrast, silencing gl20535 resulted in maximum reductions of 16.97%, 30.20%, and 23.56% in IPS, EPS, and β-1,3-glucan contents, respectively. These phenomena in the overexpression strains were attributed to gl20535-mediated promotion of UDP-glucose synthesis in the sugar donor pathway and upregulation of the expression of glycoside hydrolase genes. The opposite trend was observed in the silenced strains. In mycelial growth studies, neither overexpression nor silencing of gl20535 affected biomass and cell wall thickness. Furthermore, the GL20535 isozyme gene gl24465 remained unaffected in gl20535-overexpressed strains but was upregulated in gl20535-silenced strains, suggesting a compensatory regulatory relationship. These findings reveal the regulatory role of GL20535 on gene expression in the GLPs synthesis pathway and deepen our understanding of GL20535 function in the polysaccharide network of edible and medicinal fungi.

Bacillus licheniformis, a generally regarded as safe (GRAS) microorganism, is extensively utilized in various industries. This bacterium is notable for its ability to metabolize a wide array of carbon substrates, with 2,3-butanediol (2,3-BD) being a primary product from glucose metabolism. This compound has significant applications in various industries. Upon glucose depletion, B. licheniformis continues to grow using 2,3-BD. However, the key genes and regulatory mechanisms involved in this catabolic pathway, particularly its interaction with central metabolism, remain largely unexplored. This study employs a combination of comparative transcriptomics and gene knockout techniques to elucidate the genes essential for 2,3-BD catabolism. Our findings indicate that when 2,3-BD serves as the sole carbon source, the biomass achieved is 2.1 times that obtained with glucose. Through transcriptomic analysis and gene knockout, we identified that the 2,3-BD dehydrogenase genes (budC, gdh, yhxC), acetoin dehydrogenase complex (acoABC), aldehyde dehydrogenase (dhaS) and isocitrate lyase (aceA) are crucial for 2,3-BD catabolism. The aco operon in 2,3-BD utilization can be induced by 2,3-BD, acetoin, and aldehyde. Significantly, by reconstructing the 2,3-BD catabolic pathway in Bacillus licheniformis, we generated a mutant strain Bacillus licheniformis (Delta acoA Delta budC) which can accumulate 41.32 g/L of the intermediate metabolite acetoin, achieving a 44.3 % glucose conversion efficiency. This finding enhances our understanding of 2,3-BD catabolism in bacteria and indicates a promising acetoin producer B. licheniformis (Delta acoA Delta budC).

Hyperbranched (3-glucan from Pleurotus tuber-regium (PTR-HBG) exhibits distinct solubility, dispersion, and prebiotic effects, positioning it as a valuable resource for sustainable biotechnological applications. This study developed a predictive model for controlled depolymerization of PTR-HBG under mild acid hydrolysis (0.1-0.2 mol/L HCl at 90 degrees C) following first-order kinetics. The derived molecular weight model, Mnt= Mn0 center dot exp ( (0.6077 center dot C' + 0.5539) center dot t - (11.123 center dot C' - 0.0463)), enables precise number-average molecular weight adjustment through acid concentration (C ') and reaction time (t), facilitating production of PTR-HBG derivatives with consistent characteristics. Structural analysis indicated that degradation proceeds through a midpoint chain scission mechanism, with initial cleavage focused on (3-1,4-glucosidic bonds in the main chain, while O-6 branching points in 1,4,6-Glcp remained intact in early stages, maintaining PTR-HBG's hyperbranched structure. Transmission electron microscopy (TEM) verified the preservation of PTR-HBG's branched topology even with molecular weight reduction, while helices gradually unwound under higher acid concentrations. In vitro fecal fermentation of depolymerized products demonstrated enhanced prebiotic efficacy, promoting beneficial gut bacteria growth and persistent short-chain fatty acid (SCFA) production over 48 h compared to untreated PTRHBG. By elucidating the controlled degradation of PTR-HBG, these findings establish a foundation for enhancing bioavailability and advancing the sustainable development of targeted HBG in prebiotic applications.

Polysaccharides derived from edible and medicinal fungi (EMF) exhibit various biological activities, rendering them useful in the pharmaceutical, nutraceutical, and food industries. However, their intricate biosynthetic pathways limit their potential, necessitating a deeper understanding of their structure-function relationships. Ganoderma lucidum has emerged as a key model for elucidating polysaccharide biosynthesis in EMF. In this paper, we present a comprehensive review of the current state of research on EMF polysaccharide biosynthesis from the perspective of structure and biosynthetic pathways, and concentrate on strategies for enhancing the precise regulation of polysaccharide production. The complexity of these structures, governed by a refined biosynthetic pathway, is crucial for determining their functional diversity. This review highlights three key strategies for enhancing polysaccharide production. Genome-scale metabolic remodeling is useful for the systematic design and optimization of pathways, while profiling metabolic remodeling regulates critical genes and metabolic nodes. The regulation of enzyme activity, particularly glycosyltransferases and glycoside hydrolases, can enhance biosynthetic efficiency. Furthermore, this review identifies several future challenges pertaining to polysaccharide synthesis. We establish a robust foundation for the advanced bio-manufacturing of polysaccharides, and provide theoretical guidance, along with empirical support, to precisely regulate polysaccharide synthesis in EMF, underscoring their significance as functional foods.

Pea albumin and globulin were isolated and purified from pea protein isolates (PPI), and hybrid nanofibers were produced using electrospinning utilizing PPI, albumin, and globulin, with the assistance of pullulan (PUL). The Osborne technique separation results revealed that PPI predominantly comprised albumin (61.94 %) and globulin (17.46 %) (w/w). The P-2 fraction, which was the primary component in albumin, isolated using anion exchange chromatography demonstrated optimal antioxidant activity, while the F-2 fraction obtained via gel filtration constituted 67.81 % of the P-2 fraction. The antioxidant capacity of the F-2 fraction was similar to 1.59 times higher than that of the P-2 fraction. Under the optimized PUL concentration, i.e., 15 % PUL solution (w/v), the resultant electrospun nanofibers with PPI and albumin exhibited greater uniformity than those derived from globulin. Nanofiber characterization results showed that the protein and PUL were effectively intertwined within the nanofibers and the thermal stability of albumin-PUL surpassed that of globulin-PUL and PPI-PUL. The globulin-PUL nanofiber exhibited a slower release of quercetin, whereas the PPI-PUL nanofiber demonstrated a greater release quantity of quercetin than the other hybrid nanofibers evaluated. These results suggest that pea protein contains bioactive components and that PPI and its constituents have significant potential for application in the fabrication of electrospun nanofibers.

Flavor compounds are key determinants of food sensory quality, originating from natural sources, processing, or artificial additives. Although physical and chemical methods can effectively enhance food flavor, microbial fermentation and enzyme catalysis technology possess good potential in food flavor regulation due to their mild reaction conditions and high safety. In addition, the high efficiency and specificity of enzymes help to shorten the production cycle and accurately regulate food flavor. This review focuses on the application and regulation mechanism of bacteria, yeast, other fungi, and mixed microbe fermentation systems in flavor production. The utilization and catalytic reaction schemes of oxidoreductases, transferases, and hydrolases in flavor regulation are also deeply explored, and suggestions for the application of microbial fermentation and enzyme catalysis technology in flavor regulation are discussed.

The enzyme D-sorbitol dehydrogenase (SLDH) facilitates the conversion of D-sorbitol to L-sorbose. While current knowledge of this enzyme class predominantly centers on Gluconobacter oxydans, the catalytic properties of enzymes from alternative sources, particularly their substrate specificity and coenzyme dependency, remain ambiguous. In this investigation, we conducted BLASTp analysis and screened out a novel SLDH (Fpsldh) from Faunimonas pinastri A52C2. The SLDH was then identified and characterized. Analysis of the purified enzyme revealed its dependence on NAD+/NADP+ and its specificity for L-sorbose production. Fpsldh demonstrated sustained catalytic activity over temperatures ranging from 27 to 37 ℃, with optimal performance observed at pH 8.0–10.0, and it exhibited no requirement for metal ions for activation. The Km of Fpsldh is 7.51 mM. Furthermore, a Bacillus licheniformis host expressing Fpsldh was engineered. The resultant whole-cell catalyst yielded 13.19 g/L of L-sorbose after 33.6 h of transformation, obviating the need for exogenous cofactors. This study enhances our understanding of the catalytic properties of the SLDH family and introduces a novel method for L-sorbose production, a compound of considerable commercial value. •New D-sorbitol dehydrogenase from Faunimonas pinastri A52C2 is characterized. •Fpsldh is not PQQ but NAD+/NADP+-dependent. •Bacillus licheniformis expressing Fpsldh can produce 13.19 g/L L-sorbose within 33.6 h.

Uridine diphosphate galactose (UDP-Gal) provides galactosyl units for active carbohydrate biosynthesis; however, limited availability and high costs hamper large-scale applications. In the two-enzyme cascade system of UDP-Gal synthesis, the pH conflict between UDP-galactose 4-epimerase (GALE) and sucrose synthase (Susy) blocks UDP-Gal production. Therefore, surface charge engineering was conducted to obtain a variant (GALEM2) with improved acid resistance. GALEM2 enzyme activity reached 214.26 ± 0.20% that of wild-type GALE at pH 6.5. Its half-life time increased by 2 h at pH 6.5, and the pH resistance range was widened effectively with local surface charge reshaping and a decreased isoelectric point. An improved flexibility of the substrate entrance enhanced the catalytic performance under acidic conditions. Cascading GALEM2 and SusyM6 yielded UDP-Gal (24.5 mM) with a space-time yield of 12 g/L/h within 1.25 h, demonstrating the robust route of short reaction time and high efficiency, for rapid UDP-Gal synthesis from readily available sucrose via cascade catalysis.

Ergothioneine (EGT) production in Pleurotus tuber-regium was significantly enhanced through Na2SeO3-induced oxidative stress, in combination with optimized carbon sources and precursor amino acids. Initially, glucose and rhamnose (20 g/L each) as dual carbon sources increased EGT synthesis. Supplementation with 0.75 mM cysteine and 1 mM methionine further promoted EGT biosynthesis by providing essential precursors. Additionally, the introduction of 0.3 mM Na2SeO3 induced oxidative stress, leading to an EGT accumulation of 4.68 mg/g dry weight (DW) in the mycelium and a yield of 35.27 mg/L, representing a 4.25-fold and 162% increases compared to the control, respectively. RNA-Seq analysis revealed that selenium exposure triggered the upregulation of genes involved in antioxidant defense, reinforcing redox homeostasis and driving EGT accumulation. This study highlights the mechanistic role of selenium in modulating antioxidant pathways, offering a promising strategy for enhancing EGT production in the nutraceutical and biotechnological fields.

Laccase, a member of the blue multicopper oxidase family, is widely distributed across diverse taxonomic groups, including fungi, bacteria, plants, and insects. This enzyme drives biocatalytic processes through the oxidation of phenolic compounds, aromatic amines, and lignin derivatives, underpinning its significant potential in the food industry, cosmetics, and environmental remediation. However, wild-type laccases face critical limitations, such as low catalytic efficiency, insufficient expression yields, and poor stability. To address these bottlenecks, this review systematically examines optimization strategies for heterologous laccase expression by fungal and bacterial systems. Additionally, we discuss protein engineering for laccase modification, with a focus on the structural basis and active-site redesign. The comprehensive analysis presented herein provides strategic suggestions for advancing laccase engineering, ultimately establishing a theoretical framework for developing high-efficiency, low-cost engineered variants for large-scale biomanufacturing and green chemistry applications.

Efficient and environmentally friendly deproteinization is crucial for obtaining structurally intact edible and medicinal fungi polysaccharides. This study entailed the development of amino-functionalized magnetic beads (Fe3O4-NH2 and Fe3O4@SiO2- NH2) as recyclable and eco-friendly adsorbents for protein removal from edible and medicinal fungi polysaccharides. Under optimized conditions (pH 7.0, 40 °C, 20 mg/mL beads, 90 min), the protein removal efficiencies reached 80.9 % and 83.1 %, respectively, with polysaccharide recoveries exceeding 83 %. Coupling the beads with two Sevag cycles further improved protein removal to 94.7 %-96.3 % while maintaining recovery ≥88 %. Adsorption kinetics followed a pseudo-second-order kinetic model, indicating a spontaneous and endothermic process. Both adsorbents retained over 84 % of their initial efficiency after ten adsorption-desorption cycles, demonstrating excellent reusability. Deproteinization efficiency varied across fungal matrices: extracellular polysaccharides presented higher removal rates (79.5 %-83.5 %) than fruiting-body polysaccharides (72.8 %-78.6 %), highlighting matrix-dependent adsorption behavior. This study presents a sustainable magnetic bead-based platform that integrates green chemistry principles with efficient separation technology, providing a scalable and environmentally responsible method for producing high-purity polysaccharides from edible and medicinal fungi.

The biosynthesis of Ganoderma lucidum polysaccharides (GLPs) begins with the nucleotide sugar precursor synthesis. Guanosine diphosphate (GDP)-mannose pyrophosphorylase (GMP) specifically catalyzes the conversion of mannose-1-phosphate to GDP-mannose. However, the specific role of GMP in GLP synthesis remains unclear. Therefore, we investigated the function of the glgmp gene in mycelial growth, polysaccharide production, and the transcriptional regulation of key genes involved in GLP biosynthesis through genetic manipulation. A 2131 bp gene encoding a 50.93 kDa protein was cloned, and the overexpression and silencing transformed strains were constructed. The expression level of the glgmp gene influenced mycelial growth and pellet size. Overexpression of the glgmp gene boosted the production of high-mannose polysaccharides. The monosaccharide composition of polysaccharides changed significantly, with the proportion of mannose reaching a maximum of 13.28 % and 11.64 %. This was accompanied by a decrease in glucose percentage and an increase in galactose and fucose percentages. At the transcriptional level, overexpressing the glgmp gene upregulated the expression of phosphomannose isomerase and phosphomannomutase genes, which play critical roles in GDP-mannose synthesis. This promoted GDP-mannose supply and subsequently increased the mannose percentage. Upregulation of glgmp also increased the gene expression involved in the synthesis of GDP-fucose and uridine diphosphate (UDP)galactose, resulting in increased ratios of fucose and galactose. In contrast, the glgmp silencing strains exhibited opposite effects, confirming these findings. This study provides a valuable reference for the efficient production of GLPs, inspiring a comprehensive exploration of the polysaccharide biosynthesis pathway.