As a member of ectomycorrhizal fungi, Tricholoma matsutake has a symbiotic relationship with its host, Pinus densiflora. To cultivate T. matsutake artificially, the co-cultivation of T. matsutake mycelia and bacteria from shiro was introduced. In this study, bacteria were isolated from soil samples in Bonghwa-gun, and seven bacterial isolates (B22_7_B05, B22_7_B06, B22_7_B07, B22_7_B08, B22_7_B10, B22_7_B13, and B22_7_B14) promoted the growth of T. matsutake mycelia (147.48, 232.11, 266.72, 211.43, 175.17, 154.62, and 177.92%, respectively). Sequencing of the 16S rRNA region of the isolated bacteria was performed. B22_7_B05 and B22_7_B10 were identified as Bacillus toyonensis, B22_7_B06 and B22_7_B08 as Paenibacillus taichungensis, B22_7_B07 and B22_7_B14 as P. gorilla, and B22_7_B13 as P. odorifer. These bacterial isolates were associated with the shiro community and are expected to contribute to the cultivation of T. matsutake.

Tricholoma matsutake is a traditional favorite food in East Asia, cultivated in fairy rings called “shiro,” which are found near Pinus densiflora. For effective artificial cultivation of Tri. matsutake, microorganisms from symbiotic fairy rings are co-cultivated. In this study, one bacterial isolate (Y22_B35) and two fungal isolates (Y22_F64 and Y22_F68) displayed growth-promoting effects on Tri. matsutake mycelium (158.47, 125.00, and 122.26% enhanced growth, respectively). For identification, 16S rRNA or ITS regions from the microorganisms¡¯ genomes were sequenced. Other sequences, including BenA, CaM, and RPB2 were sequenced in the fungal isolates. The bacterial isolate Y22_B35 was identified as Bacillus cereus. Y22_F64 and Y22_F68 were identified as Umbelopsis nana and Aspergillus parvulus, respectively. To identify the effects of the dominant microorganisms on Tri. Matsutake cultivation, metagenomic analyses were performed. Discovery of these Tri. matsutake mycelium growth-promoting microorganisms and metagenomics analyses are expected to contribute to our understanding of Tri. matsutake fruiting body growth and construction of biomimicry.

Mushroom-based vegan meat has thus far been used as a food for humans instead of pets. However, based on its texture and nutritional content, it is considered suitable for processing into pet treats. In the present study, we developed a prototype dog chew with a sweetening coating added to a fungal mycelium mat obtained by culturing the Basidiomycetous fungus Trametes orientalis. The palatable coating applied to the mycelium mat by plasticizing the mat with glycerol improved the taste and aroma of the existing mat, and the dog consumed it without difficulty. Future improvements may include a softening process to reduce the chewiness level and a procedure to reduce the crude fiber content. Mycelium-mat-based dog chews, manufactured using eco-friendly materials and processes that are not harmful to the environment are expected to enter the market as eco-friendly alternatives to conventional pet treats. Controlling their physical properties require further study.

To cultivate pine mushroom (Tricholoma matsutake) artificially, co-cultivation with microorganisms has been introduced. Here, experiments were performed to assess the growth-promoting effect of bacteria on T. matsutake mycelia. Bacteria were isolated from soil samples collected in Yangyang County, Korea. Four of the bacterial isolates (Y22_B06, Y22_B11, Y22_B18, and Y22_B22) exhibited a growth-promoting effect on T. matsutake mycelia (154.67%, 125.91%, 134.06%, and 158.28%, respectively). To analyze the characteristics of the bacteria, especially the antifungal activity, -amylase and cellulase activity assays were performed. In comparison with the controls, the isolated bacteria exhibited low -amylase and cellulase activity. 16S rRNA gene sequencing was performed to identify the four bacterial isolates. The isolates belonged to the Terrabacteria group and were identified as Microbacterium paraoxydans, Paenibacillus castaneae, Peribacillus frigoritolerans, and P. butanolivorans. These bacterial isolates are expected to have contributed to the growth promotion of T. matsutake mycelia and the artificial cultivation of T. matsutake.

This study was conducted to develop a renewable and sustainable bio-material to replace polystyrene (EPS) in fungal-mycelium-based composite using agricultural by-products. Four mushrooms (Ganoderma lucidum, Fomitella fraxinea, Phellinus linteus, and Schizophyllum commune) were cultured in an oak sawdust plus rice bran substrate to select the mushroom with the best growth. The mycelia of G. lucidum showed the best growth. To investigate the optimal mixing ratio with spent mushroom substrate (SM) and oak sawdust (OS), samples were prepared by mixing SM and OS at ratios of 50%:50%, 60%:40%, and 80%:20% (w/w). Each substrate was then inoculated with G. lucidum. G. lucidum showed the best mycelial growth of 140.0 mm in the substrate with SM and OS mixed at a 60%:40% ratio. It was also found that the substrate with SM and OS mixed at a 60%:40% ratio had the best handling properties. The compressive strength of mycelial materials inoculated with G. lucidum was in the range of 300–302 kgf mm-1, and the materials were four times stronger than polystyrene materials. These results indicate that substrates comprising spent mushroom substrate mixed with oak sawdust can be successfully upcycled to mycelium-based composite materials using G. lucidum. This represents a sustainable approach.

In this study, the protein content and functional changes in soybeans cultured with Phellinus linteus HN00K9 were analyzed. P. linteus HN00K9 was cultured on soybeans. The crude protein content in soybeans cultured with HN00K9 (PMS) was 41.99%, which was higher than that in soybeans not cultured with the mushroom (UCS). The total free amino acid content in PMS increased to 39,963 mg/100 g, which was higher than that in UCS (36,817 mg/100 g). In particular, in PMS, glutamic acid accounted for 18.5% of the total amino acids at 7,413 mg/100 g. The total polyphenol content in PMS was 2.66 mg GAE/g, which was more than 45% higher than the amount in UCS (1.45 mg GAE/g). Additionally, PMS showed a DPPH radical scavenging activity of 33.3%, which was 3 times higher than that exhibited by UCS (11.5%), reflecting its high antioxidant content. Therefore, the PMS in this study has potential for use as a functional food material.

The importance of biocomposites has increased owing to the changes in global consumption trends and rapid climate change. Technologies using mushroom mycelium cultivation, and molding methods for mycelial application have gained attention as potential strategies for producing eco-friendly composites. Currently, mushroom mycelia are used as raw materials for food and cosmetics; however, research on their utilization as biocomposite materials is limited. Therefore, the potential for the development of mushroom mycelium-related products and technologies is high. This review analyzes the domestic and international patent application trends related to the technologies for composite (packaging, insulation, adhesives, and leather) and food (substitute for meat) materials using mushroom mycelium, as an eco-friendly biocomposite material, to provide objective patent information that can further research and development (R&D) in this field.

In this study, we aimed to compare the mycelial growth of Pleurotus ostreatus after medium supplementation with various amino acids at different concentrations to select the optimal medium nutrient composition for mycelial growth. The mycelial growth of P. ostreatus was investigated after adding four amino acids (tryptophan, threonine, methionine, and lysine) at 0.5% or 1% to the medium.The rate of P. ostreatus mycelial growthwas faster in the potato dextrose agar (PDA) medium supplemented with threonine at 0.5% or 1% than that of the control, whereas it was inhibited by tryptophan treatment. Supplementation of sawdust mediumwith all amino acids, except tryptophan, at 0.5% did not alter the mycelial growth, compared to the controls. However, addition of any amino acid to sawdust medium at a higher concentration (1%) inhibited the mycelial growth. The laccase acitivity of P. ostreatus mycelium cultured in PDA medium was the highest when threonine was added, and the lowest when tryptophane was added, consistent with the results of the mycelial growth. Therefore, the addition of threonine, methionine, or lysine to PDA medium at a concentration of 0.5-1%was effective for increasing the mycelial growth of P. ostreatus; however, it inhibited mycelial growth insawdust medium, suggesting that the effects of amino acids dependedon the medium nutrient composition.

To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

After liquid culture of Phellinus baumii (P. baumii) mycelium (LPBM) was prepared, LPBM was fractionated into A∼E fraction (A; hot-water extract of liquid culture including mycelia, B; crude polysaccharide of A, C; hot-water extract of mycelia, D; crude polysaccharide of C, and E; crude polysaccharide of culture broth) to evaluate for possibility as functional materials with immunostimulatory activity. In macrophage stimulatory activity, E fraction as postbiotics significantly increased secretion of NO and IL-12 from RAW 264.7 cells. Next, when the splenocytes of C3H/HeN mice were primary cultured, E fraction showed significantly mitogenic activity with enhancing mitogen-related cytokines (IFN-γ and TNF-α) production from splenocyte. E fraction also potently stimulated GM-CSF production from Peyer’s patch cells as well as Peyer’s patch-mediated bone marrow cell proliferation. In addition, the immunostimularoy E fraction contained neutral sugar (73.8%), uronic acid (10.6%), protein (7.8%), and polyphenol (7.5%), and mainly consisted of glucose (39.1%), galactose (21.7%), mannose (11.1%), galacturonic acid (9.9%), and arabinose (8.9%) as component sugars. In conclusion, it was demonstrated that postbiotics including exopolysaccharide fractionated from liquid culture of the P. baumii mycelium could enhanced immunostimulatory activity.

This study investigated the culture characteristics of Cryptoporus volvatus, whichis grow naturally in Korea, to determine the suitable environmental conditions for its cultivation. The physiological characteristics of the mycelia were assessed according to the cultivation conditions, to determine the optimal conditions for artificial cultivation. The visual characteristics of the hyphae of Cryptoporus volvatus KACC52303 included an irregular and uneven surface and a fuzzy or cotton-like texture. Under the microscope, its microstructure showed pre-chlamydospore formation, but no clamps were seen. The appropriate culture temperature was found to be a medium/high temperature of approximately 25–30oC, and the optimal pH was found to have a wide range from weakly acidic (pH 4) to neutral (pH 7). In the optimal nutrient source experiment, hyphal growth was shown to be fair in a mixed medium with 2.5% dextrin as the carbon source and 0.1% yeast extract as the organic nitrogen source. Among the various amino acids, organic acids, and inorganic salts tested, the fastest hyphal growth was observed in the presence of leucine, acetic acid or gluconic acid, and KCl or KH2PO4, respectively. The column test showed that the best mycelial growth occurred in a mixed medium of 80% pine sawdust, 10% rice bran, and 10% corncob sawdust.

Bio-based alternative leathers may be produced from biomass fiber, protein polymers, bacterial cellulose, and mushroom mycelia. Of these components, mushroom mycelia are of greatest interest. In this study, the potential of Fomes fomentariusas a mushroom mycelial mat was confirmed, and the optimal strain for the development of the mycelial mat was determined. Moreover, the quality of the mycelial mat was improved by identifying an efficient culture method to increase productivity. Mutant strains whose independence was verified were obtained by treatment with gamma irradiation under various conditions. Biofilm formation by the resulting strains was examined in sawdust and liquid media and the characteristics of the biofilms were analyzed. The biofilm of the mutant strains showed results that were similar to or better than the biofilms of longevity and cypress mushrooms. These findings are expected to be utilized in future research aimed at discovering new biomaterials using mushroom mycelia.

This study was conducted to investigate the growth and antioxidant activity of Pleurotus ostreatus mycelia grown in medium supplemented with Aronia. The diameter and dry weight of the mycelia were increased in the medium supplemented with Aronia compared with the untreated medium. The total polyphenol content of mycelia grown in medium supplemented with Aronia and untreated medium was 6.54 mg GAE/g and 5.77 mg GAE/g, respectively. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of mycelia grown in medium supplemented with Aronia and untreated medium was 61.81% and 49.65%, respectively. Moreover, the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity of mycelia grown in medium supplemented with Aronia and untreated medium was 59.83% and 52.66%, respectively. These results confirmed that P. ostreatus mycelial growth and antioxidant activity were increased when Aronia was added to the culture medium.

Eco-friendly materials, such as alternative vegan materials using various fungal resources, are being actively researched to reduce environmental pollution and facilitate a healthy lifestyle. The fungal mycelium-based mushroom mycelium mat is one such emerging material. In this study, the commonly used mushroom mycelium culture method was modified to reduce the time required to produce the mycelium mat, lower the possibility of contamination, and improve the properties and quality of the mat. Shortening the period required for the previously used primary bag culture and secondary mat production culture. A culture method in which the bag culture was omitted was attempted using a mycelium mutated by gamma irradiation to the mycelium of Trametes orientalis. In addition, various nutrients were added to the fungal solution to observe the change in physical properties of the fungal mat. High-quality mycelium mats were produced in the experimental group containing 1.5% CaCO3 in sawdust medium, and the period was also reduced by more than 10 days compared to the existing production method. In the future, for mass producing mycelium mats, additional selection of medium components and optimization of culture conditions are essential.

To investigate the industrial availability of liquid fermentation (PL-ferment) by Phellinus linteus mycelium as a postbiotics for the inhibition of inflammation, PL-ferment was fractionated into culture supernatant (CS), hot-water extract (HW) from PL-ferment, EtOH-precipitate (CP) fractionated from HW, and the dialysate (DCP) of CP. Compared to the other fractions, DCP which is expected to contain exopolysaccharide (EPS) as the major component, significantly decreased the production of NO, IL-6, and MCP-1 in LPS-induced RAW 264.7 cells, and IL-6 and IL-8 in TNF-α and IFN-γ-induced HaCaT cells. The general component analysis results showed that no significant difference in components was observed between the fractions, whereas sugar composition analysis revealed that DCP had decreased glucose and increased mannose contents compared to the other fractions. This suggests that mannose played an important role in the anti-inflammatory activity of the active fraction, DCP. Molecular weight distribution analysis revealed that DCP was mainly composed of low-molecular-weight material-removed high-molecular-weight polysaccharides of 18–638 kDa, suggesting that EPS originated from P. linteus EPS. In conclusion, our results suggest that the DCP of P. linteus mycelium fermentation using the anti-inflammatory activity could be used industrially as postbiotic material.

This study observed the anti-inflammatory effect of the polysaccharide derived from the mycelium of Tremella fuciformis in mice with colitis induced with dextran sulfate sodium (DSS). The experimental groups were normal, DSS, DSS-TFL50, DSS-TFH100, and suflasalazine. Body weights, colon lengths, and organ weights were measured, and the plasma level of pro-inflammatory cytokine and mRNA and protein expression in colon tissue were analyzed. Body weight loss, a symptom of DSS-induced colitis, was suppressed by DSS-TF and the speed of weight recovery proceeded rapidly. In addition, DSS-TF showed a significant inhibitory effect on the decrease of colon length typically caused by colon damage. TNF-α, IL-6 and IL-1β cytokine levels in plasma were reduced in DSS-TF and positive control groups. TNF-α, COX-2 and IL-1β mRNA expression in colon tissue were inhibited in DSS-TF and positive control, and it was significantly different from that of the DSS group. The protein expression of inflammation-related genes (IL-6, TNF-α and COX-2) in the colon tissue was significantly increased by DSS compared to that of the normal group, but by DSS-TFL50, DSS-TFH100 and sulfasalarin decreased. In conclusion, the polysaccharide derived from the mycelium of Tremella fuciformis showed the anti-inflammatory effect on DSS-induced colitis in mice.

본 연구는 아미노산 스포츠 음료를 개발하기 위하여 음료 베이스로 영지버섯, 차가버섯, 상황버섯복합버섯 균사 체 추출물을 사용하여 제조하였다. 차가버섯, 상황버섯. 영지버섯은 베타글루칸 다량 함유하여 면역증강작용과 같 은 인체에 유용한 성분을 다량 함유하고 있다고 다수의 연구에서 보고하고 있다. 선행연구에 의하면 3종의 버섯 균사체 추출물의 항산화 활성을 비교한 결과, 매우 높은 활성을 보여 스포츠 음료나 건강 음료의 재료로 적합할 것으로 판단되어 본 연구에 사용하였다. 3종의 버섯 균사체를 배양한 후 80℃, 60min 가열 추출하여 아미노산 음료 베이스로 사용하였고, 아르기닌을 포함한 필수 아미노산과 레몬, 자몽, 다크체리 농축액을 혼합하여 각각 3, 5, 7% 로 첨가하여 스포츠 음료를 제조하고 그 특성을 비교하였다. 개발 스포츠 음료의 고형분 함량은 농도에 비례해서 약 3.0~7.2brix로 측정되었고, pH도 3.94~4.37범위였다. 이들 스포츠 음료의 항산화 활성을 비교한 결과는 다음과 같다. 총 페놀 함량은 레몬·자몽(LG)첨가 음료가 약 726.74~798.87mg/L로 높게 측정되었고, 체리(C)첨가 음료는 585.03~794.73mg/L이었다. DPPH 라디칼 소거능을 비교한 결과, LG7이 71.57%로 체리 첨가 음료보다 가장 높게 측 정되었다. 반면, 총 플라보노이드 함량을 비교한 결과, C7이 53.35mg/L로 높았으나, 다른 시료들도 유의적으로 높아, 총 플라보노이드 함량은 과일농축액의 종류와 농도에 크게 영향을 받지 않은 것으로 관찰되었다. 따라서 본 연 구에서 개발된 스포츠 음료는 생리활성이 높게 규명된 재료를 사용하였고, 필수 아미노산을 첨가하여 제조한 제품으로 스포츠 활동을 한 후 단백질 보충용 음료로써의 기능을 할 수 있을 것으로 예상된다.