• Title/Summary/Keyword: Medicinal fungus

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In vitro Antitumor Activity of Ergosterol Peroxide Isolated from Cordyceps militaris on Cancer Cell Lines from Korean Patients (Cordyceps militaris로부터 분리한 Ergosterol Peroxide의 한국인 암세포주에 대한 항암작용)

  • Kim, Ha-Won;Kim, Young-Ho;Cai, Xing Fu;Nam, Kyong-Suk;Lee, Seung-Jung;An, Hye-Suk;Jeong, Eun-Ho;Yun, Seung-Hyun;Sung, Su-Kyong;Lee, Sung-Jin;Hyun, Jin-Won
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.61-66
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    • 2001
  • Entomopathogenic fungus of Cordyceps militaris is famous of its medicinal efficacies. An antitumor compound was purified from the n-hexane extract of artificially cultivated fruiting bodies of C. militaris and identified as ergosterol peroxide $(5{\alpha},\;8{\alpha}-epidioxy-24(R)-methylcholesta-6,22-dien-3{\beta}-o1,\; C_{28}H_{44}O_3)$ mainly by $^1H\;and\;^{13}C-NMR$ spectroscopic techniques. When the antitumor activity of ergosterol peroxide was measured against 3 tumor cell lines from Korean cancer patients, it showed the most strong activity against gastric cancer SNU-l cell line 3 days after treatment. The 50% inhibitory concentrations $(IC_{50})$ of ergosterol peroxide 6 days after treatment were $75.8{\mu}g/ml$ for human gastric SNU-1 tumor cell line, $39.7{\mu}g/ml$ for human colorectal SNU-C4 tumor cell line and $32.7{\mu}g/ml$ for human hepatoma SNU-354 cell line.

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Various physiological effects from fruiting body extracts of Phellinus baumii (장수진흙버섯 자실체 추출물의 다양한 생리활성 효과)

  • Yoon, Ki Nam;Lee, Tae Soo
    • Journal of Mushroom
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    • v.18 no.3
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    • pp.260-267
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    • 2020
  • Phellinus baumii, a white-rot fungus, has been used for centuries as folk medicine in China, Japan, and Korea. This study aimed to evaluate the in vitro anti-diabetic, and anti-cholinesterase, and in vivo anti-inflammatory effects of the fruiting bodies of P. baumii. The methanol (ME) and hot water (HE) extracts (2.0 mg/mL) of P. baumii fruiting bodies suppressed α-amylase activity, exactly 61.33%, and 65.00%, respectively; of note, acarbose, the positive control, inhibited 93.33% of the α-amylase activity. Moreover, the ME and HE (2.0 mg/mL) inhibited 89.67% and 91.00%, respectively, of the activity of α-glucosidase activity, whereas the same concentration of acarbose suppressed 84.67% of the α-glucosidase activity. The ME and HE (1.0 mg/mL) also inhibited 96.05% and 94.58%, respectively, of the acetylcholinesterase (AChE) activity; galanthamine, the positive control, led to an inhibition of 81.12%. The butyrylcholinesterase (BChE) activity was also inhibited by ME and HE (1.0 mg/mL; 91.05% and 82.27%, respectively); of note, the same concentration of galanthamine suppressed 81.12% of the BChE activity. The production of NO in LPS-induced RAW 264.7 macrophages was significantly suppressed by both ME and HE treatments. Importantly, the carrageenan-activated rat hind-paw edema was significantly reduced 2-6 h after ME administration (50 mg/mL). Taken together, the results suggest that the fruiting bodies of P. baumii have α-amylase, α-glucosidase, α-cholinesterase, and anti-inflammatory activities, and, therefore, may be good natural sources for the promotion of human health.

Induction of Apoptotic Cell Death by Aqueous Extract of Cordyceps militaris Through Activation of Caspase-3 in Human Hepatocarcinoma Hep3B Cells (Hep3B 간암세포에서 Caspase-3 활성화를 통한 동충하초 열수추출물의 Apoptosis 유도에 관한 연구)

  • Kim, Kyung-Mi;Park, Cheol;Seo, Sang-Ho;Hong, Sang-Hoon;Lee, Won-Ho;Choi, Yung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.714-720
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    • 2008
  • Cordyceps militaris is a medicinal fungus which has been used for patient suffering from cancer in Oriental medicine. It was previously reported that C. militaris extracts are capable of inhibiting tumor growth and inducing apoptosis; however, the anti-poliferative effects of human cancer cells have been poorly understood. In this study, to elucidate the anti-cancer mechanisms of human cancer cells by treatment with aqueous extract of C. militaris (AECM), we investigated the anti-proliferative effects of AECM in human hepatocarcinoma Hep3B cells. AECM treatment inhibited the growth of Hep3B cells and induced the apoptotic cell death in a concentration-dependent manner such as formation of apoptotic bodies and increased populations of apoptotic-sub G1 phase. The induction of apoptosis by AECM was connected with a proteolytic activation of caspase-3 and caspase-8. and concomitant degradation of poly (ADP-ribose) polymerase (PARP) and ${\beta}$-catenin proteins. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited AECM-induced apoptosis demonstrating the important role of caspase-3 in the bserved cytotoxic effect. Taken together, these findings suggest that AECM-induced inhibition of human hepatocarcinoma cell proliferation is associated with the induction of apoptotic cell death via activation of caspase-3 and C. militaris may have therapeutic potential in human cancer.

Development of New Strains of Wolfiporia cocos for Sclerotium Formation by 2-Way Cross-Breeding (이원교배에 의해 균핵 형성하는 복령 균주 개발)

  • Ka, Kang-Hyeon;Kim, Suyeon;Park, Mi-Jeong;Jeong, Yeun Sug;Ryoo, Rhim;Jang, Yeongseon;Choi, Jong-Woon;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.49 no.3
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    • pp.405-412
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    • 2021
  • Wolfiporia cocos is an important medicinal fungus that has been used in regions of Northeast Asia including Korea, Japan, and China. W. cocos is classified in Korea into two types (red bokryeong and white bokryeong) based on the internal colors (yellow orange-pale pink and white) of the sclerotium. Generally, the W. cocos type cultivated on farms produces white sclerotium. In this study, we endeavored to select strains that form sclerotium in sawdust medium using 2-way cross-breeding among two cultivated strains and three wild strains. Monospores were isolated from the fruiting bodies of cultivated and wild strains on potato dextrose agar. Thirty-nine strains of 338 hybrid strains isolated formed sclerotia with white or yellow colors upon culture for 3 months in Pinus densiflora sawdust medium. Selection for sclerotium forming strains using sawdust culture follows a very simple and easy procedure that is presented for the first time in this paper. We plan to test selected strains in the field to aid in developing new varieties for the future.

The Antifungal Effect of Rhus verniciflua Stokes against Metarhizium anisopliae on the Edible Insect, Protaetia brevitarsis (Coleoptera) (흰점박이꽃무지에 발생하는 병원성 곰팡이 Metarhizium anisopliae에 대한 옻나무 추출액의 항진균 효과)

  • Kim, Nang-Hee;Song, Myung-Ha;Kim, Eunsun;Kim, Yongsoon;Park, Kwan-Ho;Kim, Sunyoung
    • Korean Journal of Ecology and Environment
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    • v.53 no.4
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    • pp.374-379
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    • 2020
  • The white-spotted flower chafer (Protaetia brevitarsis) larva is one of the insects widely-used for edible and medicinal purposes in Eastern Asia. As a result of increasing demand for P. brevitarsis, massrearing systems in domestic farms have become necessary. However, the mass-rearing of larvae under confined rearing conditions could provide conditions unsuitable for preventing entomopathogenic diseases. Metarhizium anisopliae is the strongest fatal entomopathogenic fungus against P. brevitarsis. For inhibition of M. anisopliae, we used a Rhus verniciflua Stokes extract that has antifungal components. We investigated the inhibitory effect of the R. verniciflua extract at 1%, 5%, and 10% concentrations. The results showed that a 1% R. verniciflua extract added to sawdust produced a significantly low P. brevitarsis mortality rate. Moreover, extract-treated groups were heavier and had a shorter larval period than those of the untreated group. Consequently, we suggest that using an R. verniciflua extract can reduce the P. brevitarsis fatality rate from entomopathogenic fungi (e.g. M. anisopliae), resulting in more effective mass-rearing systems for P. brevitarsis.

Effects of Elevated CO2 Concentration and Increased Temperature on the Growth of Gastrodia elata Blume, Parasitic Medicinal Plant (CO2 농도와 온도 변화가 기생 약초인 천마의 생육반응에 미치는 영향)

  • Cho, Kyu-Tae;Lee, Soo-In;Jang, Rae-Ha;Park, Jae-Hoon;You, Young-Han
    • Korean Journal of Environment and Ecology
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    • v.31 no.6
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    • pp.557-563
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    • 2017
  • Gastrodia elata (Orchidaceae) is a perennial parasitic plant that has a unique lifestyle of being in a symbiotic interaction with fungi. The underground root of Gastrodia is used for medicinal treatment to cure high blood pressure, stroke, leukemia, headaches, and especially neurasthenia. This study is intended to investigate the effect of the global warming on the ecological responses of Gastrodia. We cultivated the small tuber of Gastrodia with the oak wood lot and mulberry fungus under control (ambient $CO_2$ concentration + ambient temperature, ACAT), temperature treatment (ambient $CO_2$ concentration+elevated temperature, ACET), and $CO_2$+temperature treatment (elevated $CO_2$ concentration+elevated temperature, ECET). The elevated $CO_2$ concentration was about twice in the ambient air while the elevated temperature was about $2^{\circ}C$ higher than the control group. And then we observed the growth and production of reproductive organs and the underground root. The observation showed that the number of flower stalk was highest at ACET and lowest at ECET. The flower stalk was longest at ACET and shortest at ECET. The inflorescence was longest at ACAT and shortest at ECET. The seed capsule was heaviest at ACET and lightest at ECET. The aboveground biomass was highest at ACET and lowest at ECET. The number of rhizomes was highest at ACET and lowest at ECET. The total rhizome biomass was highest at ACET and lowest at ECET. The average rhizome biomass was highest at ACET and lowest at ECET. The results showed that the growth of Gastrodia increased because of more active growth of Gastrodia elata when only the temperature increased and decreased when both $CO_2$ concentration and temperature increased, indicating the poor growth of Gastrodia elata under the global warming condition. Therefore, the Gastrodia elata plantation should be maintained at the temperature of $20-25^{\circ}C$ and not be exposed to a high $CO_2$ concentration.

Extracts of Allium fistulosum Attenuates Pro-Inflammatory Action in the Lipopolysaccharide-Stimulated BV2 Microglia Cells (Lipopolysaccharide에 의한 BV2 세포의 염증 반응에 대한 파 추출물의 저해 활성)

  • Park, Shin-Hyoung;Kim, Jung-In;Jeong, Yong-Kee;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.21 no.6
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    • pp.796-804
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    • 2011
  • Microglia are central nervous system (CNS)-resident professional macrophages that function as the principal immune cells responding to pathological stimulations in the CNS. Activation of microglia, induced by various pathogens, protects neurons and maintains homeostasis in the CNS, but severe activation causes inflammatory responses secreting various neurotoxic molecules such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines. Allium fistulosum, a member of the onion family, is mainly cultivated for consumption, as well as medicinal use in Oriental medicine. It has been reported that A. fistulosum has various biological effects such as anti-oxidant, anti-platelet aggregation, anti-fungus and anti-cholesterol synthesis, however there has been no research about the anti-inflammatory effects of A. fistulosum extracts. In this study, it was undertaken to explore the functions of A. fistulosum as a suppressor of neuronal inflammation by using BV2 microglia cells. As a result, it was found that four kinds of extracts of A. fistulosum effectively reduced the expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels, and also attenuated pro-inflammatory cytokines such as tumor necrosis alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and interleukin-6 (IL-6) at the mRNA level in BV2 stimulated by lipopolysaccharide (LPS). In addition, the extracts of A. fistulosum attenuated the release of NO markedly, as well as resulting in slight decreases of TNF-${\alpha}$ and IL-6 production, the effects of which were most significant when treated with ethyl alcohol extract from the whole A. fistulosum. In conclusion, the data indicated that the anti-inflammatory actions of A. fistulosum against BV2 microglia cells is through the down-regulation of iNOS, COX2 and pro-inflammatory cytokines such as TNF-${\alpha}$ and IL-6, and these effects are expected to help in the protection of nerve tissues by suppressions of neuronal inflammation in various neurodegenerative diseases.

Collection, Identification and Hepatic Effect of Native Cordyceps militaris (새로운 번데기 동충하초의 수집, 동정 및 간기능에 미치는 효과)

  • Lee, Ki-Won;Nam, Byung-Hyouk;Jo, Wool-Soon;Oh, Su-Jung;Kang, Eun-Young;Cui, Yong;Lee, Jae-Yun;Cheon, Sang-Cheol;Jeong, Min-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.34 no.1
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    • pp.7-14
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    • 2006
  • Entomopathogenic fungus Cordyceps militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. In this study, we investigated the effect of the extract of C. militaris (MPUN8501), which was identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA, on the function of liver. C. militaris powder was extracted using hot water extracts method as time, volume and temperature and using method as differential polarity of organic solvent. Each fraction was tested for the improvement of hepatic enzyme alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activity. The BuOH extracts (CME) had highest activity which was used for the test of toxicity and efficacy of C. militaris. The enhancing effect of CME on the activity of ADH and ALDH was much more than medicine, drink, natural tea etc. Thus CME promoted the resolution of alcohol and acetaldehyde in rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters (levels of sGOT and sGPT) and histological observation. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintaining normal hepatic activity as a functional health food.

Genetic Diversity and Phylogenetic Relationship in Korean Strains of Lentinus lepideus Based on PCR Polymorphism (PCR 다형성 분석에 의한 한국산 잣버섯의 유전적 다양성 및 유연관계)

  • Lee, Jae-Seong;Cho, Hae-Jin;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Lee, Yun-Hae;Jang, Myoung-Jun;Ju, Young-Chul;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.105-111
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    • 2010
  • Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 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.

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