Extraction of chitin-glucan complex from shiitake (Lentinula edodes) fruiting bodies using natural deep eutectic solvents and its prebiotic potential.

Chitin-glucan complex (CGC) is an emerging novel prebiotic with numerous physiological activities in amelioration of clinical manifestations. In the present work, natural deep eutectic solvent (NADES), ultrasonication, and submerged fermentation using probiotic microorganisms were deployed for the extraction of CGC from Shiitake fruiting bodies. CGC obtained through non-ultrasonication assisted fermentation employing Lactiplantibacillus plantarum exhibited maximum polysaccharide yield (27.86 ± 0.82 % w/w). However, based on antioxidant potential, NADES combination of urea: glycerol (1:1 M ratio) was selected for further characterization. The rheological behavior of CGC under optimized conditions showed shear thinning property in both 0.1 M NaCl and salt-free solution. FTIR, 1H-(1D), and 2D 1H1H Homonuclear NMR spectra displayed distinctive patterns associated with ß-glycosidic linkage and ß-d-glucopyranose sugar moiety. XRD profiles of CGC exhibited characteristic peaks at 2θ = 23°, 25°, and 28° with corresponding hkl values of (220), (101), and (130) lattice planes, respectively. Enhanced radical scavenging activities were noticed due to the triple helical structure and anionic nature of CGC. CGC exhibited potential prebiotic activity (prebiotic score 118-134 %) and short chain fatty acids liberation (maximum 9.99 ± 0.41 mM by Lactobacillus delbrueckii). Simulated static in-vitro digestion demonstrated that CGC withstands acidic environment of gastric phase, which indicated its suitability for use as a prebiotic in nutraceutical-enriched food products.

Metabolomic differences between non-hydrothermal treated water-soluble (WSPs) and hydrothermally treated water-insoluble (WIPs) Maitake polysaccharides fermented by Lactobacillus acidophilus and L. plantarum.

Bacterial Metabolite through a fermentation process is a growing trend and a promising alternative for use as functional components. Non-hydrothermal water-soluble (WSPs) and hydrothermally treated water-insoluble (WIPs) Maitake polysaccharides were fermented with Lactobacillus acidophilus (LA) and Lactobacillus plantarum (LP). Chemical composition analysis indicated that Maitake polysaccharides contained 58.22 ± 1.35 % total sugar and 31.46 % ß-glucan, essential for metabolites production. 6-glucanase was used to degrade the WIPs, and hydrothermally treated WIP fibers exhibited smooth microstructure. Hence, the LA and LP bacteria investigated the potential fermented metabolic activities and differences between WSPs(Sp1)and WIP(Sp3) Maitake polysaccharides using LC-MS, and 887 metabolites were identified. Using Venn, Partial least squares discriminant analysis (PLS-DA), VIP Metabolites, and other multivariate statistical analysis methods, metabolites were expressed differently in all samples. Due to hydrothermal processing, WIP induced the highest growth of LA and LP, with an abundance of isocitrate metabolites. Furthermore, 50 metabolite correlations were identified, leading to the classification of 6 distinct metabolic groups. Thus, the study offers the initial comprehensive analysis of metabolites in Lactobacillus-fermented Maitake polysaccharides, aiding in understanding its metabolic interactions and facilitating progress in food engineering research.

Formation and non-covalent interactions of binary and ternary complexes based on ß-casein, Lentinus edodes mycelia polysaccharide, and taxifolin.

Polyphenols, polysaccharides, and proteins are essential nutrients and functional substances present in food, and when present together these components often interact with each other to influence their structure and function. Proteins and polysaccharides are also excellent carrier materials for polyphenols. In this context, this study investigated the non-covalent interactions between taxifolin (TAX), Lentinus edodes mycelia polysaccharide (LMP), and ß-casein (ß-CN). ß-CN and LMP spontaneously formed nanocomplexes by hydrogen bonds and van der Waals forces. The quenching constant and binding constant were (1.94 ± 0.02) × 1013 L mol-1 s-1 and (3.22 ± 0.17) × 105 L mol-1 at 298 K, respectively. The altered conformation of ß-CN, resulting from the binding to LMP, affected the interaction with TAX. LMP significantly enhanced the binding affinity of TAX and ß-CN, but did not change the static quenching binding mode. The binding constant for ß-CN-TAX was (3.96 ± 0.09) × 1013 L mol-1, and that for the interaction between TAX and ß-CN-LMP was (32.06 ± 0.05) × 1013 L mol-1. In summary, ß-CN-LMP nanocomplexes have great potential as a nanocarrier for polyphenols, and this study provides a theoretical foundation for the rational design of non-covalent complexes involving LMP and ß-CN, both in binary and ternary configurations.

Novel Catechol O-methyltransferases from Lentinula edodes Catalyze the Generation of Taste-Active Flavonoids.

Due to the increasing demand for natural food ingredients, including taste-active compounds, enzyme-catalyzed conversions of natural substrates, such as flavonoids, are promising tools to align with the principles of Green Chemistry. In this study, a novel O-methyltransferase activity was identified in the mycelium of Lentinula edodes, which was successfully applied to generate the taste-active flavonoids hesperetin, hesperetin dihydrochalcone, homoeriodictyol, and homoeriodictyol dihydrochalcone. Furthermore, the mycelium-mediated OMT activity allowed for the conversion of various catecholic substrates, yielding their respective (iso-)vanilloids, while monohydroxylated compounds were not converted. By means of a bottom-up proteomics approach, three putative O-methyltransferases were identified, and subsequently, synthetic, codon-optimized genes were heterologously expressed in Escherichia coli. The purified enzymes confirmed the biocatalytic O-methylation activity against targeted flavonoids containing catechol motifs.

Preparation and characterization of water vapor-responsive methylcellulose-polyethylene glycol-400 composite membranes and an indication of freshness of shiitake mushrooms.

Intelligent packaging with freshness indication capability can help consumers purchase fresh food. However, current research primarily focuses on carbon dioxide-sensitive intelligent packaging, with limited research on water vapor-sensitive indication packaging. In this study, the water vapor-sensitive indicator membrane was prepared and used to determine the freshness of mushrooms. The results of this study showed that the water permeability of the indicator membrane decreased from 33.17 % to 21.59 % with the increase of Polyethylene glycol-400(PEG-400) content in methylcellulose(MC) membrane, and the contact angle of the indicator membrane increased from 87 % to 98 % with the addition of PEG-400. The addition of plasticizer PEG-400 increased the hydrophobicity of the indicator film, which could be attributed to the improvement of the molecular arrangement and crystallinity of the indicator film by the addition of PEG-400. After encountering water, the transparency of the indicator membrane changes from completely opaque (white) to transparent. Addition of PEG-400 reduces the rate of change in the transparency of the indicator membrane. The indicator membrane was successfully used to indicate the freshness of mushrooms and effectively reflected the freshness of mushrooms during storage. This technology could be applied to measure the freshness of other foods.

Extraction of Lentinus Edodes Polysaccharides with Ultrasound Enhanced with Deep Eutectic Solvent and Their Structural Characterization and Antioxidant Activity.

Ultrasound extraction (UE) enhanced with deep eutectic solvent (DES) was used to extract Lentinus edodes polysaccharides. Box-Behnken design (BBD) was applied to investigate the influences of water content (10-90 %), solid-liquid solvent (1 : 10-1 : 50 g/mL), time (4-12 min), temperature (40-80 °C) and ultrasonic power (100-500 W) on the yield of Lentinus edodes polysaccharides. The optimal extraction conditions were ultrasonic power of 300 W, extraction time of 8 min, water content of 80 %, a solid-liquid ratio of 1 : 30 g/mL and a temperature of 60 °C, respectively. The highest extraction yield of Lentinus edodes polysaccharide was 10.17 % under optimal conditions. The results of FT-IR, SEM, and monosaccharide composition confirmed that the extracts possessed the characteristics of polysaccharides. In addition, the polysaccharides obtained with the UE enhanced with DES method exhibited higher antioxidant activities than the polysaccharides extracted with the UE method and HWE method. This extraction method can further expand the production efficiency and structural diversity of Lentinus edodes polysaccharides and meet the supply and demand relationship. It can be foreseen that this method can be applied to the extraction of more active substances.

Analysis of Volatile Aroma Components in Different Parts of Shiitake Mushroom (Lentinus edodes) Treated with Ultraviolet C Light-Emitting Diodes Based on Gas Chromatography-Ion Mobility Spectroscopy.

Further assessment of ultraviolet C light-emitting diode (UVC-LED) irradiation for influencing shiitake mushrooms' (Lentinus edodes) volatile and sensory properties is needed. In this study, a comparison of UVC-LED irradiation treatment on the flavor profiles in various parts of shiitake mushrooms was conducted using gas chromatography-ion mobility spectrometry (GC-IMS) and sensory analysis. Sixty-three volatile compounds were identified in shiitake mushrooms. The fresh shiitake mushrooms were characterized by the highest values of raw mushroom odors. After UVC-LED treatment, the content of C8 alcohols decreased, especially that of 1-octen-3-ol, while the content of aldehydes increased, especially the content of nonanal and decanal. The score of fatty and green odors was enhanced. For fresh samples, the mushroom odors decreased and the mushroom-like odors weakened more sharply when treated in ethanol suspension than when treated with direct irradiation. The fruit odors were enhanced using direct UVC-LED irradiation for fresh mushroom samples and the onion flavor decreased. As for shiitake mushroom powder in ethanol suspension treated with UVC-LED, the sweaty and almond odor scores decreased and the vitamin D2 content in mushroom caps and stems reached 668.79 µg/g (dw) and 399.45 µg/g (dw), respectively. The results obtained from this study demonstrate that UVC-LED treatment produced rich-flavored, quality mushroom products.

Edodin: A New Type of Toxin from Shiitake Mushroom (Lentinula edodes) That Inactivates Mammalian Ribosomes.

Ribosome-inactivating proteins (RIPs) are a group of proteins with rRNA N-glycosylase activity that irreversibly inhibit protein synthesis and consequently cause cell death. Recently, an RIP called ledodin has been found in shiitake; it is cytotoxic, strongly inhibits protein synthesis, and shows rRNA N-glycosylase activity. In this work, we isolated and characterized a 50 kDa cytotoxic protein from shiitake that we named edodin. Edodin inhibits protein synthesis in a mammalian cell-free system, but not in insect-, yeast-, and bacteria-derived systems. It exhibits rRNA N-glycosylase and DNA-nicking activities, which relate it to plant RIPs. It was also shown to be toxic to HeLa and COLO 320 cells. Its structure is not related to other RIPs found in plants, bacteria, or fungi, but, instead, it presents the characteristic structure of the fold type I of pyridoxal phosphate-dependent enzymes. Homologous sequences have been found in other fungi of the class Agaricomycetes; thus, edodin could be a new type of toxin present in many fungi, some of them edible, which makes them of great interest in health, both for their involvement in food safety and for their potential biomedical and biotechnological applications.

Detoxification of tetracycline and synthetic dyes by a newly characterized Lentinula edodes laccase, and safety assessment using proteomic analysis.

Fungal laccase has strong ability in detoxification of many environmental contaminants. A putative laccase gene, LeLac12, from Lentinula edodes was screened by secretome approach. LeLac12 was heterogeneously expressed and purified to characterize its enzymatic properties to evaluate its potential use in bioremediation. This study showed that the extracellular fungal laccase from L. edodes could effectively degrade tetracycline (TET) and the synthetic dye Acid Green 25 (AG). The growth inhibition of Escherichia coli and Bacillus subtilis by TET revealed that the antimicrobial activity was significantly reduced after treatment with the laccase-HBT system. 16 transformation products of TET were identified by UPLC-MS-TOF during the laccase-HBT oxidation process. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that LeLac12 could completely mineralize ring-cleavage products. LeLac12 completely catalyzed 50 mg/L TET within 4 h by adding AG (200 mg/L), while the degradation of AG was above 96% even in the co-contamination system. Proteomic analysis revealed that central carbon metabolism, energy metabolism, and DNA replication/repair were affected by TET treatment and the latter system could contribute to the formation of multidrug-resistant strains. The results demonstrate that LeLac12 is an efficient and environmentally method for the removal of antibiotics and dyes in the complex polluted wastewater.

Lentinus edodes mycelium polysaccharide inhibits AGEs-induced HUVECs pyroptosis by regulating LncRNA MALAT1/miR-199b/mTOR axis and NLRP3/Caspase-1/GSDMD pathway.

A novel Lentinus edodes mycelia polysaccharide (LMP) prepared in our laboratory has been identified to be effective in inhibiting the damage of islet ß cells induced by glucose toxicity. However, whether it can effectively alleviate the pyroptosis of human umbilical vein endothelial cells (HUVECs) induced by advanced glycation end products (AGEs) remains unclear. Bioinformatics and cell biology techniques were used to explore the mechanism of LMP inhibiting AGEs-induced HUVECs damage. The results indicated that AGEs significantly increased the expression of LncRNA MALAT1, decreased cell viability to 79.67 %, increased intracellular ROS level to 248.19 % compared with the control group, which further led to cell membrane rupture. The release of LDH in cellular supernatant was increased to 149.42 %, and the rate of propidium iodide staining positive cells increased to 277.19 %, indicating the cell pyroptosis occurred. However, the above trend was effectively retrieved after the treatment with LMP. LMP effectively decreased the expression of LncRNA MALAT1 and mTOR, promoted the expression of miR-199b, inhibited AGEs-induced HUVECs pyroptosis by regulating the NLRP3/Caspase-1/GSDMD pathway. LncRNA MALAT1 might be a new target for LMP to inhibit AGEs-induced HUVECs pyroptosis. This study manifested the role of LMP in improving diabetes angiopathy and broadens the application of polysaccharide.

Effects of combined drying techniques and cellulase hydrolysis on the nutritional value and sensory properties of shiitake mushrooms (Lentinus edodes).

Dried shiitake mushrooms offer rich nutritional value and unique sensory properties, prompting further investigation. The effects of different drying techniques (hot air drying (HAD), infrared hot air drying (IRHAD), pulsed vacuum drying (PVD), vacuum freeze drying (VFD), and natural drying (ND)) combined with enzymatic hydrolysis on the release of flavor compounds and nutrients from shiitake mushrooms were explored. The combination of HAD with cellulase hydrolysis yielded notably high levels of umami amino acids (5.4723 ± 0.1501 mg/g) and 5'-nucleotides (4.0536 ± 0.0062 mg/g), and superior volatile flavors. Combined with cellulase hydrolysis, IRHAD achieved the highest level of total sugars (6.57 ± 0.34 mg/mL), VFD resulted in the greatest soluble protein content (153.21 ± 0.23 µg/mL), PVD yielded the highest total phenolics content (93.20 ± 0.41 µg GAE/mL), and ND produced the maximum reducing sugar content (5.79 ± 0.13 mg/mL). This study addresses crucial gap in the post-drying processing of shiitake mushrooms, offering valuable insights for further product development of shiitake mushrooms.

Near-gapless genome and transcriptome analyses provide insights into fruiting body development in Lentinula edodes.

Fruiting body development in macrofungi is an intensive research subject. In this study, high-quality genomes were assembled for two sexually compatible monokaryons from a heterokaryotic Lentinula edodes strain WX1, and variations in L. edodes genomes were analyzed. Specifically, differential gene expression and allele-specific expression (ASE) were analyzed using the two monokaryotic genomes and transcriptome data from four different stages of fruiting body development in WX1. Results revealed that after aeration, mycelia sensed cell wall stress, pheromones, and a decrease in CO2 concentration, leading to up-regulated expression in genes related to cell adhesion, cell wall remodeling, proteolysis, and lipid metabolism, which may promote primordium differentiation. Aquaporin genes and those related to proteolysis, mitosis, lipid, and carbohydrate metabolism may play important roles in primordium development, while genes related to tissue differentiation and sexual reproduction were active in fruiting body. Several essential genes for fruiting body development were allele-specifically expressed and the two nuclear types could synergistically regulate fruiting body development by dominantly expressing genes with different functions. ASE was probably induced by long terminal repeat-retrotransposons. Findings here contribute to the further understanding of the mechanism of fruiting body development in macrofungi.

In vitro evaluation of methanol extracts of edible fungi Pleurotus ostreatus and Lentinula edodes against Rhyssomatus nigerrimus Fahraeus.

The study aimed to evaluate the insecticidal activity of extracts of edible mushrooms Pleurotus ostreatus and Lentinula edodes against Rhyssomatus nigerrimus. Methanol:water extracts (70:30) of P. ostreatus and L. edodes were made and evaluated in two in vitro tests (exposure and immersion toxic effect) against R. nigerrimus. Subsequently, the toxicity test of the extracts against Artemia salina was evaluated. These extracts were subjected to colorimetric tests and gas chromatography-mass spectrometry. The results showed a mortality effect against R. nigerrimus of 50% for the P. ostreatus 2 extracts at a concentration of 20% in the immersion test. Likewise, in the toxic effect test, 90% mortality was observed after five days of exposure to a concentration of 10%. On the other hand, for the toxicity test, the extract that showed the values with the highest mortality against A. salina was P. ostreatus, starting with 80% mortality at 100µg/mL. The functional groups present in the extracts were saponins, coumarins, and alkaloids. Furthermore, the presence of more than 7 compounds in the mushroom extracts evaluated is reported. This study demonstrates the insecticidal activity of P. ostreatus and L. edodes fungal extracts and indicates the importance of using different in vitro tests to elucidate the mechanism of action for future studies.

The Tumor Growth Inhibitory Effect of a Standardized Extract of Cultured Lentinula edodes Mycelia Using Patient Derived Xenograft Model.

Patient derived xenograft (PDX) is a powerful tool to confirm pharmacological efficacy in non-clinical studies for the development of various drugs including anti-cancer agents and therapeutic research. A standardized extract of cultured Lentinula edodes mycelia, a product name AHCC® is produced by Amino Up Co., Ltd. (Sapporo, Japan). In this study, we investigated the inhibitory effect of AHCC® on the growth of tumor PDX in Super SCID (severe combined immunodeficiency) mice. Effects of AHCC® and BCG administration on the growth of renal cancer PDX implanted in Super SCID mice were evaluated by PDX growth curve. Tendency for the effects on the growth of renal cancer PDX in Super SCID by administration of AHCC® and BCG before implanting the PDX were demonstrated. The effects of the oral administration of AHCC® on the growth of renal, invasive and non-invasive breast cancer PDX in Super SCID mice were studied. In Super SCID mice transplanted with renal cancer PDX, AHCC® significantly suppressed tumor proliferation from the day 48 to 83 after transplantation. In two types of breast cancer PDX, tendency of the growth inhibitory effects of AHCC® were shown by PDX growth curve. Significant inhibitory effect was found at only one time point for during proliferation in each PDX. Super SCID-PDX model has the potential to be a useful tool to investigate for the effect of functional foods.

The improvement effects of Lentinus edodes powder marination on sous vide cooked chicken patties: Physicochemical attributes, oxidative properties and flavor characteristics.

The improvement effects of Lentinus edodes powder (LEP) marination with different concentrations (0, 6-14 %) on physicochemical, oxidative and flavor quality of chicken patties were evaluated. Greater pH, redness, yellowness, water holding capacity and their strong correlations were observed in LEP-marinated samples. Changed water distribution, inhibited lipid oxidation and enhanced protein oxidation occurred through LEP marination. The highest gel strength and resilience and the lowest hardness and chewiness were obtained in 10 % LEP-marinated sample. Meanwhile, taste activity values of amino acids and saltiness peaked and umami rose in this sample. 124 volatiles were detected and 16 compounds were simultaneously detected by gas chromatography-ion mobility spectrometry and gas chromatography-mass spectrometry. Hexanal, 1,2,4-trithiolane and 1-hexanol were considered as the key differential aroma-active compounds according to odor activity values and chemometric analysis. This study confirmed LEP as a prospective ingredient to improve the quality of meat products.

Anti-inflammatory and immunomodulatory properties of polysaccharides in mushrooms.

Mushrooms are distinguished as important food-containing polysaccharides possessing potent anti-inflammatory and immunomodulating properties. These compounds belong mostly to polysaccharides that are mostly ß-D-glucans. Among them, ß-1,3-glucan with ß-1,6 side chains of glucose residues, has more important roles in their properties. In this review, we have introduced polysaccharides mainly from Lentinula edodes and Pleurotus citrinopileatus with anti-inflammatory and immunomodulating properties. In addition, the mechanisms of activation of their physiological properties and signal cascade are also reviewed.

Hepatoprotection of a Standardized Extract of Cultured Lentinula edodes Mycelia against Liver Injury Induced by Ischemia-Reperfusion and Partial Hepatectomy.

A standardized extract of cultured Lentinula edodes mycelia (ECLM, AHCC®) has been shown to have beneficial effects on organ metabolism. ECLM has been indicated to have liver protective properties by suppressing inflammatory responses. The pathogenesis of hepatic ischemia-reperfusion injury is thought to involve the induction of inflammatory mediators. However, whether ECLM affects inflammatory mediators caused by warm hepatic ischemia-reperfusion injury and partial hepatectomy (HIRI+PH) has not been clarified. In this study, we evaluated the protective effects of ECLM against liver damage caused by HIRI+PH. Rats were fed a normal diet (HIRI+PH) or a normal diet with 2% ECLM (HIRI+PH and ECLM) for ten days, then the liver and duodenal ligament were clamped and subjected to 15 min of hepatic ischemia. After 70% hepatectomy, the inflow occlusion was released, and liver and blood samples were collected at 3, 6, and 24 h. The effect of ECLM on mortality induced by 30 min of ischemia and hepatectomy was evaluated. The results showed that ECLM attenuated pathological liver damage, including apoptosis, in the rats treated with HIRI+PH, and decreased serum aminotransferase activity; ECLM decreased mRNA levels of the inflammation-related genes inducible nitric oxide synthase and C-X-C motif chemokine ligand 1, and increased mRNA levels of interleukin 10, an anti-inflammatory cytokine; ECLM increased hepatocyte growth factor mRNA levels and Ki-67 labeled nuclei in the liver at 24 h; ECLM significantly reduced HIRI+PH-induced mortality. In conclusion, ECLM may prevent HIRI+PH-induced liver injury in part by suppressing various inflammatory responses and promoting liver regeneration.

Photoregulation of the biosynthetic activity of the edible medicinal mushroom Lentinula edodes in vitro.

The findings of the study demonstrate the impact of low-intensity laser and quasi-monochromatic light on the biosynthetic activity of the edible medicinal fungus L. edodes during submerged cultivation. An artificial lighting installation based on matrices of light-emitting diodes (LED) emitting light at 470 nm (blue), 530 nm (green), 650 nm (red), and argon gas laser (488 nm) was used. Irradiation with blue and red LED and laser led to a shortening of the lag phase by 2 days and an increase in the mycelial mass. Irradiation with laser light resulted in the highest mycelial mass yield (14.1 g/L) on the 8th day of cultivation. Irradiation in all used wavelength ranges caused an increase in the synthesis of both extracellular and intracellular polysaccharides. Laser light at 488 nm and LED at 470 nm proved to be the most effective. Irradiation with red, green, and blue laser light caused an increase in the total amount of fatty acids in the mycelial mass compared to the control. A significant distinction in qualitative composition was observed: short-chain acids C6‒C12 compounds were produced under red light irradiation, whereas long-chain C20‒C24 were formed under green light irradiation. The most significant changes in the aromatic profile of the mycelial mass and culture liquid were recorded upon irradiation with green light. The content of aromatic components increased 24.6 times in the mycelial mass and 38.5 times in the culture liquid. The results suggest the possibility of using low-intensity quasi-monochromatic light for targeted regulation of L. edodes biosynthetic activity.

Peach palm shells (Bactris gasipaes Kunth) bioconversion by Lentinula edodes: Potential as new bioproducts for beef cattle feeding.

This paper aims to develop and assess the in vitro effects on ruminal fermentation and greenhouse gas parameters of new bioproducts for beef cattle diets, carried out by solid-state fermentation of peach palm shells colonized by Lentinula edodes (SSF) and after Shiitake mushroom cultivation in axenic blocks (SMS). In vitro experiments were performed to assess the in vitro gas production, digestibility, and fiber degradation of formulated total diets. Bioproducts presented high ß-glucans (9.44---11.27 %) and protein (10.04---8.35 %) contents, as well as similar digestibility to conventional diets. SMS diet had the lowest methane and carbon dioxide (19.1 and 84.1 mM/g OM) production, and the SSF diet presented lower carbon dioxide production (98.9 mM/g OM) than other diets, whereas methane was similar. This study highlighted a sustainable use of byproducts for beef cattle diets, promising for digestibility, nutritional value, ß-glucans incorporation, and environmental impact mitigation, favoring the circular bioeconomy.

Effects of different cooking methods on proximate composition, digestion characteristics, and antioxidant activity of Lentinus edodes.

Lentinus edodes (SM) are highly appreciated by the food industry together with consumers for nutrition and flavour. It is hypothesized the results of our study can indicate to the suitable cooking method for maximal nutrient retention, this study investigated the effects of six cooking methods on the proximate composition, digestive properties, and antioxidant activities of SM in vitro simulated digestion. The results revealed that the ash and protein contents of cooked samples were reduced excluding steaming and roasting, and likewise minerals during deep-frying and boiling (e.g., Mg from 1080.07 to 629.03 mg/kg, deep-frying). A conspicuous rise in fat and energy was found in deep-frying and stir-frying. Steaming retained more reducing sugar (3.80 mg/mL). Roasting improved bioaccessibility of most amino acids (e.g., 82.61%, Asp). Using oil as a medium was associated with higher antioxidant activities using water. Steaming and roasting were better for preserving the nutrient composition of cooked Lentinus edodes.