• Journal of Mushroom
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• v.22 no.1
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• pp.1-7
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• 2024

Mushrooms play crucial roles as reservoirs of naturally occurring bioactive compounds. Among these, Cordyceps militaris is significant because of its well-established reputation for organoleptic excellence and positive health effects, which have led to its widespread commercialization. In contrast, the key properties of Paecilomyces variotii, an ectomycorrhizal symbiont, has received limited attention. In alignment with current research trends, the study of the mycelia and culture media of these mushrooms hold promise in identifying potential sources of valuable bioactive compounds. In the present study, we investigated C. militaris and P. variotii for their phenolic acids and sterols, assessing antioxidant capacity, anti-inflammatory effects, and anti-proliferative activity. Interestingly, P. variotii mycelia exhibited higher concentrations of ergosterol and phenolic compounds, with comparable levels observed in the fruiting bodies, along with superior antioxidant activity compared to that of C. militaris. In contrast, C. militaris mycelia demonstrated anti-inflammatory effects (which were absent in P. variotii mycelia) and cytotoxicity comparable to, and at times exceeding, that of its fruiting bodies (in contrast to P. variotii). In addition, the species analyzed in this study displayed variations in growth rates and mycelial production, which merit consideration for potential future applications and further study.

• KSBB Journal
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• v.25 no.1
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• pp.47-54
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• 2010

The mycelial dispersed growth of Cordyceps sinensis was optimized in submerged batch culture at initial pH of 5.0, 150 rpm, and $25^{\circ}C$. The morphological data showed much more dispersed growth of C. sinenesis at initial pH of 5.0. Also, projected area, main hyphal length and number of tips for the mycelial growth of initial pH 5.0 were higher than those of other initial pHs. The industrial medium for mycelial production of C. sinensis was determined to be molasses of 100 g and crushed brewery yeast of 10 g per liter as carbon and nitrogen sources, respectively. With these culture conditions, the maximum production of mycelia was approximately 30.0 g per liter by batch culture in 5-liter jar fermenter with no controlled pH. This result suggests that large-scale mycelia production of C. sinensis may be possible in submerged batch culture. The hot water extract of mycelia from C. sinensis was mainly composed of 83.0% carbohydrate, 11.8% protein, 1.9% lipid, and 2.4% ash and there were present glucose, mannose, galactose, and arabinose as molar ratio of 8.79 : 2.59 : 1.34 : 1.0 in the carbohydrate, respectively. In the experiment using spleen cell and macrophage, the extract showed potent mitogenic and immuno-stimulating activities and among various components, an important factor that contribute to the immunological activities was turned out to be carbohydrate moiety.

• Journal of Mushroom
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• v.18 no.3
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• pp.189-200
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• 2020

Pleurotus djamor var. roseus is an edible mushroom isolated from the wild and cultivated on paddy straw substrates. The present study was carried out to compare the nutritional composition and antioxidant properties of P. djamor var. roseus at different growth stages (primordia, basidiomata, and mycelia). The protein content was is in the range of 31.48 to 35.50 g/100g dw. The crude fiber content ranged from 8.0 to 14.60 g, and that of total carbohydrates ranged from 44.75 to 48.90 g. Sodium, magnesium, and calcium reached the maximum levels in basidiomata, and selenium was detected in basidiomata and mycelia (0.47 - 0.22 mg/Kg). The amino acid profile showed that all essential and nonessential amino acids and glycine showed maximum levels in basidiomata and 15.98 ± 0.01 g/100g. The fatty acid profile showed the presence of saturated and unsaturated fatty acids; the unsaturated fatty acid content was maximum in all of the samples, ranging from 76 - 40.41%. The total phenol and flavonoid contents as well as the scavenging (DPPH), ferric thiocyanate (FTC), and thiobarbituric acid (TBA) activities in the context of methanol and water extracts from primordia, basidiomata, and mycelium were determined. Among them, basidiomata and mycelial methanol extracts exhibited significant antioxidant activity. Overall, these findings show that P. djamor var. roseus can be used as a functional food for daily consumption.

• 한국균학회소식:학술대회논문집
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• 2016.05a
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• pp.19-20
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• 2016

Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

• Weed & Turfgrass Science
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• v.2 no.2
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• pp.202-206
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• 2013

Biocontrol potential of an isolate of Helicosporium spp. against Rhizoctonia solani, Fusarium oxysporium and Phytophthora drechsleri was evaluated in vitro and in vivo. A selected biocontrol agent designated as Helicosporium 0635BP strongly inhibited growth and lysed mycelium of Rhizoctonia solani and Fusarium oxysporium on PDA. Autoclaved culture filtrate of the agent also completely inhibited growth of the turfgrass large patch pathogen, R. solani AG2-2 IV at the concentration of 50 ml $L^{-1}$. The pathogen was killed when dipped under the 20% filtrate for four hours or 50% for one hr. In a field trial, plots applied with the crude or times diluted culture filtrate showed 100% control efficacy of the turfgrass large patch as a chemical applied for a comparison. Results indicated that Helicosporium 0635BP is a promising biocontrol agent on control of the turfgrass large patch disease and its culture filtrate contained unknown heat suitable antifungal substance (s). Further studies on mass production, purification and identification of the unknown compound (s) are in progress for practical use.

• Journal of Mushroom
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• v.9 no.4
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• pp.145-154
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• 2011

Shiitake mushroom (Lentinula edodes) is usually cultivated on the oak log. Log cultivation of this mushroom is getting difficult to get oak logs and has a weak point of its long cultivation period. Recently sawdust cultivation is getting increase. It is important to make mycelia browning on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. The period for mycelia browning is so long that the sawdust cultivation of Shiitake mushroom can not spread well into the mushroom farms. The development of methods for the rapid mycelia browning is quite required. In this article we would like to find cultural characteristics of collected strains and to see the correlation with mycelial browning. Mycelial growth in the media was different according to kinds of media and strains. The optimal temperature on mycelial growth was $20-25^{\circ}C$. Browning patterns of mycelium under 200 Lux seemed to be used for a key to differentiate the strains for sawdust cultivation. Browning period was 30-40 days in the agar media and 70-100 days in the sawdust bag cultivation. When we considered the productivity and the other characteristics, ASI 3046 is the best for the bag cultivation. Significance between mycelial growth and browning was not accepted, but that of mycelial growth between on PDA and sawdust was accepted. Browning period on the PDA and sawdust showed a strong relationships. These results suggested that the browning habits could not be depend on the difference of media, but on their own properties. To select the strain showed fast browning can be done by using agar media for saving time.

• Applied Biological Chemistry
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• v.21 no.3
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• pp.150-184
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• 1978

Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.