• 제목/요약/키워드: Reactive nitrogen species

검색결과 118건 처리시간 0.031초

치매병태(癡呆病態)모델에서 천마(天麻)의 신경세포(神經細胞) 손상(損傷) 보호효과(保護效果) (Protective Effect of Gastrodia Elata on Neuronal Cell Damage in Alzheimer's Disease)

  • 정영수;강재현;박세환;권영미;김근우;구병수
    • 동의신경정신과학회지
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    • 제21권2호
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    • pp.125-140
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    • 2010
  • Objectives : The purpose of this study is to examine from various angles the protective effect of Gastrodia elata Blume (GEB) against nerve cell death induced by $\beta$-amyloid by using the cell line SH-SY5Y, which is commonly utilized for toxicity testing in nerve cells, and to find out its mechanism of action. Methods : To begin with, as a result of assessing the rate of cell survival by employing MTT reduction assay, the treatment with $\beta$-amyloid at different concentrations caused cytotoxicity, which was inhibited by preprocessing GEB extract. In addition, after $\beta$-amyloid was processed with the cell SH-SY5Y, apoptosis progressed, which was reduced effectively by processing GEB extract. Results : When cytotoxicity was caused by using hydrogen peroxide, a representative ROS, in order to examine the antioxidant effect of GEB, its protective effect was also observed. Apart from ROS, reactive nitrogen species (RNS) are also known to play a crucial role in nerve cell death. The treatment with the NO donor SNAP increased the production of nitric oxide and the expression of iNOS, which was also inhibited by GEB extract. Meanwhile, as an attempt to find out the mechanism of action explaining the antioxidant effect, the intracellular antioxidant enzyme expressions were measured by RT-PCR, which showed that GEB extract increased the expressions of heme oxygenase-1, GAPDH and $\gamma$-glutamate cysteine ligase. Lastly, GEB extract had a protective effect against impaired memory induced by scopolamine in animal models (in vivo). Conclusions : These findings indicate that GEB has a protective effect against the death of cranial nerve cells, suggesting possibilities for the prevention and treatment of AD.

익수지선단(益壽地仙丹)이 Streptozotocin으로 유발된 당뇨병 흰쥐의 신기능, 활성산소, 활성질소 및 Polyol Pathway에 미치는 영향 (Effects of Iksujisundan on Renal Function, Peroxynitrite Scavenging Activity and Polyol Pathway in Streptozotocin-induced Diabetic Rats)

  • 전창민;정지천
    • 대한한의학회지
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    • 제28권1호통권69호
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    • pp.237-248
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    • 2007
  • Objectives : Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Iksujisundan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Iksujisundan on renal function, peroxynitrite(ONOO-) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. Methods : The crushed Iksujisundan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 87.8g. Iksujisundan extract was orally administreted at 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin(60mg/kg). The effects of Iksujisundan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical(${\cdot}$O2-), nitric oxide(NO) and ONOO-, and also the enzyme activities involved in the polyol pathway. Results : The effects of Iksujisundan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and indulin levels, creatinine and BUN levels, total cholesterol and triglyceride lavels, and HDL-cholesterol levels were all shown to be good enough to prevent and cure the diabetes and its complications. Iksujisundan inhibited the generation of ${\cdot}$O2-,NO and ONOO- in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats were reversed toward natural activities. Conclusions : Iksujisundan might inhibit the development of diabetes and its complications by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover kidney function.

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Streptozotocin으로 유발된 흰쥐의 당뇨병성 신증에서 가미구기환동환(加味枸杞還童丸)이 Oxidative Stress 및 Polyol Pathway에 미치는 영향 (Effects of Gamigukihwandong-hwan on Renal Function, Oxidative Stress and Polyol Pathway in Diabetic Nephropathy Rats)

  • 정형철;정지천
    • 동의생리병리학회지
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    • 제21권3호
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    • pp.671-678
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    • 2007
  • Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Gamigukihwandong-hwan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Gamigukihwandong-hwan on renal function, peroxynitrite (ONOO$^-$) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. The crushed Gamigukihwandong-hwan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 74.95 g. Gamigukihwandong-hwan extract was oral-administered 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin (60 mg/kg). The effects of Gamigukihwandong-hwan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical (${\cdot}O_2^-$), nitric oxide (NO) and ONOO$^-$, and also the enzyme activities involved in polyol pathway. The Effects of Gamigukihwandong-hwan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and insulin levels, creatinine and BUN levels, total cholesterol and triglyceride levels, and HDL-cholesterol levels were all shown to be good enough to cure and prevent the diabetes and its complications. Gamigukihwandong-hwan inhibited the generation of ${\cdot}O_2^-$, NO and ONOO$^-$ in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats, whereas the ones in the Gamigukihwandong-hwan administered group among the streptozotocin-induced diabetic rats were reversed toward the natural activities. Gamigukihwandong-hwan might inhibit the development of diabetic nephropathy by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover the function of kidney.

맥문동(麥門冬)의 증숙(蒸熟)에 따른 항산화 효능 및 고지혈증 유발 흰쥐에 대한 효능 연구 (Effects of Steaming Process on Liriopis Tuber to Antioxidant Activities and Hyperlipidemia Induced Rats.)

  • 구가람;이현인;김수지;신미래;이아름;박해진;노성수;서영배
    • 대한본초학회지
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    • 제33권5호
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    • pp.89-103
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    • 2018
  • Objectives : This study is aimed to compare the changes in Antioxidative capacity of Liriopis Tuber by steaming process and to compare the effects in hyperlipidemia induced rats fed high cholesterol diet between Simvastatin and Liriopis Tuber by steaming process. Methods : The SD rats were divided into six groups: normal diet (Nor), high cholesterol diet (Veh), high cholesterol diet plus Simvastatin 5 mg/kg (Sim), high cholesterol diet plus LT0 extract 200 mg/kg (LT0), high cholesterol diet plus LT6 extract 200 mg/kg (LT6) and high cholesterol diet plus LT9 extract 200 mg/kg (LT9). We compared the total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) contents and reactive oxygen species (ROS) from each serums. Protein expression in liver tissues related to antioxidant and cholesterol was analyzed. Results : The Antioxidant activity of Liriopis Tuber increased by steaming process. In vivo, TC, TG, LDL-c, atherogenic index (AI) and cardiac risk factor (CRF) decreased and HDL-c increased with increasing steaming frequency. aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, and blood urea nitrogen (BUN) decreased with increasing steaming frequency. ROS decreased only in LT9, and SOD, catalase and glutathione peroxidase (GPx) increased with increasing steaming frequency. phospho-AMP-activated protein kinase (p-AMPK) increased and sterol regulatory element-binding protein 2 (SREBP-2), Phospho-Acetyl-CoA Carboxylase (p-ACC) and HMG-CoA reductase (HMGCR) decreased with increasing steaming frequency. Liver staining showed a decrease in hepatic fat accumulation of LT9. LT9 showed significant results in all experiments. Conclusions : LT9 showed significance of anti-lipid effect and improved fatty liver of hyperlipemia induced rats fed on high cholesterol diet, In conclusion, LP9 can be effectively used for the treatment of hyperlipidemia.

Anti-fatigue effect of tormentic acid through alleviating oxidative stress and energy metabolism-modulating property in C2C12 cells and animal models

  • Ho-Geun Kang;Jin-Ho Lim;Hee-Yun Kim;Hyunyong Kim;Hyung-Min Kim;Hyun-Ja Jeong
    • Nutrition Research and Practice
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    • 제17권4호
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    • pp.670-681
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    • 2023
  • BACKGROUND/OBJECTIVES: Oxidative stress is caused by reactive oxygen species and free radicals that accelerate inflammatory responses and exacerbate fatigue. Tormentic acid (TA) has antioxidant and anti-inflammatory properties. Thus, the aim of present study is to determine the fatigue-regulatory effects of TA in H2O2-stimulated myoblast cell line, C2C12 cells and treadmill stress test (TST) and forced swimming test (FST) animal models. MATERIALS/METHODS: In the in vitro study, C2C12 cells were pretreated with TA before stimulation with H2O2. Then, malondialdehyde (MDA), lactate dehydrogenase (LDH), creatine kinase (CK) activity, tumor necrosis factor (TNF)-α, interleukin (IL)-6, superoxide dismutase (SOD), catalase (CAT), glycogen, and cell viability were analyzed. In the in vivo study, the ICR male mice were administered TA or distilled water orally daily for 28 days. FST and TST were then performed on the last day. In addition, biochemical analysis of the serum, muscle, and liver was performed. RESULTS: TA dose-dependently alleviated the levels of MDA, LDH, CK activity, TNF-α, and IL-6 in H2O2-stimulated C2C12 cells without affecting the cytotoxicity. TA increased the SOD and CAT activities and the glycogen levels in H2O2-stimulated C2C12 cells. In TST and FST animal models, TA decreased the FST immobility time significantly while increasing the TST exhaustion time without weight fluctuations. The in vivo studies showed that the levels of SOD, CAT, citrate synthase, glycogen, and free fatty acid were increased by TA administration, whereas TA significantly reduced the levels of glucose, MDA, LDH, lactate, CK, inflammatory cytokines, alanine transaminase, aspartate transaminase, blood urea nitrogen, and cortisol compared to the control group. CONCLUSIONS: TA improves fatigue by modulating oxidative stress and energy metabolism in C2C12 cells and animal models. Therefore, we suggest that TA can be a powerful substance in healthy functional foods and therapeutics to improve fatigue.

플라즈마 보조 유기금속 화학기상 증착법에 의한 MCN(M=Ti, Hf) 코팅막의 저온성장과 그들의 특성연구 (Low Temperature Growth of MCN(M=Ti, Hf) Coating Layers by Plasma Enhanced MOCVD and Study on Their Characteristics)

  • 부진효;허철호;조용기;윤주선;한전건
    • 한국진공학회지
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    • 제15권6호
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    • pp.563-575
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    • 2006
  • Ti(C,N) 박막을 온도범위 $200-300^{\circ}C$에서 tetrakis diethylamido titanium유기금속 화합물을 전구체로 이용하여 pulsed DC 플라즈마 보조 유기금속 화학기상 증착법 (PEMOCVD)으로 합성하였다. 본 연구에서는 플라즈마 특성을 서로 비교하기 위하여 수소$(N_2)$와 헬륨/수소$(He/H_2)$ 혼합기체를 각각 운반기체로 사용하였으며 전구체 이외에 질소$(N_2)$와 암모니아$(NH_3)$ 기체를 반응기체로 사용하여 서로 다른 플라즈마 화학조건에서 얻어지는 박막내의 탄소함유량(C Content)의 변화를 비교하여 탄소가 가장 적게 함유된 저온 코팅막 합성공정을 찾으려고 하였다. 이를 위하여 증착시 서로 다른 pulsed bias 전압과 기체종류 하에서 여기된 플라즈마 상태의 라디칼종들과 이온화 경향을 in-situ optical emission spectroscopy(OES)법으로 플라즈마 진단분석을 실시하였다. 그 결과 $(He/H_2)$ 혼합기체를 $N_2$와 함께 사용할 경우 라디칼 종들의 이온화를 매우 효과적으로 향상시킴을 관찰하였다. 아울러 $NH_3$ 기체를 $H_2$ 또는 $He/H_2$ 혼합기체와 같이 사용할 경우는 CN 라디칼의 생성을 억제하여 결과적으로 Ti(C, N) 박막내의 탄소함량을 크게 낮춤을 알 수 있었고, CN 라디칼의 농도가 탄소 함유량과 많은 관련이 있음을 알았다. 이 결과는 바로 박막의 미세경도와도 연관이 되며, bias전압과 기체종류에 크게 의존하여 Ti(C, N) 박막의 미세경도가 1250 - 1760 Hk0.01 사이에서 나타났고, 최대치$(1760\;Hk_{0.01})$는 600 V bias 전압과 $H_2$$N_2$ 기체를 사용한 경우에 얻어졌다. HF(C, N) 박막 역시 tetrakis diethylamido hafnium 전구체와 $N_2/He-H_2$ 혼합기체를 이용하여 pulsed DC PEMOCVD 법으로 기판온도 $300^{\circ}C$ 이하, 공정압력 1 Torr, 그리고 bias전압과 기체 혼합비를 변화시키면서 증착하였다. 증착시 in-situ OES 분석결과 플라즈마 내의 질소종의 함유량 변화에 따라 증착속도가 크게 변화됨을 알 수 있었고, 많은 질소기체를 인입하면 질소종이 많아지지만 증착률은 급격히 감소하였고 박막내 탄소의 함량이 커지면서 막질이 비정질로 바뀌고 미세경도 또한 감소함을 알 수 있었다. 이는 in-situ 플라즈마 진단분석이 전체 PEMOCVD 공정에 있어서 대단히 중요하고, Ti(C,N)과 Hf(C,N) 코팅막의 탄소함량과 미세경도는 플라즈마내의 CH과 CN radical종의 세기에 크게 의존함을 의미한다. 그리고 Hf(C,N) 박막의 경우도 Ti(C,N) 박막의 경우와 유사하게 최대 미세경도값$(2460\;Hk_{0.025})$이 -600 V bias 전압과 10% 질소기체 혼합비를 사용한 경우에 얻어졌고, 이는 박막이 주로(111) 방향으로 성장됨에 기인한 것으로 사료된다.

PM2.5로 자극한 폐상피세포의 $NF{\kappa}B$ 활성화에 NO의 역할 (Role of NO in Activation of $NF{\kappa}B$ by PM2.5 in Lung Epithelial Cells)

  • 김경아;남혜윤;문제혁;정진숙;임영
    • Tuberculosis and Respiratory Diseases
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    • 제52권6호
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    • pp.616-626
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    • 2002
  • 연구배경 : PM2.5는 호흡기질환의 악화 및 호흡기 질환에 의한 사망률과 밀접하게 관계가 있다. PM2.5에 의한 호흡기 염증반응의 악화가 그 원인으로 생각되나 염증반응의 조절기전은 확실하지 않다. 본연구에서는 PM2.5에 의한 폐장의 염증반응을 조절하는데 중요한 역할을 하는 전사인자인 $NF{\kappa}B$의 활성화 기전을 알아보아 염증 조절기전에 대한 이해를 높이고자 시도되었다. 방 법 : 폐상피세포주인 A549 세포에서 PM2.5 자극에 의한 $NF{\kappa}B$의 활성화 및 ROS, 그리고 RNS 분비를 관찰 하였다. iNOS 억제제인 L-NIL가 $NF{\kappa}B$ 활성화에 미치는 영향을 관찰하였다. 결 과 : PM2.5는 A549 세포에서 $NF{\kappa}B$의 활성화를 유도 하였다. PM2.5로 A549 세포를 자극시 즉각적인 RNS의 분비는 관찰되었으나 명확한 ROS의 증가는 관찰되지 않았다. RNS 억제제인 L-NIL 처리시 $NF{\kappa}B$ 활성화는 억제되었다. 결 론 : A549 세포에서 PM2.5에 의하여 유도된 즉각적인 $NF{\kappa}B$ 활성화 과정에는 RNS가 중요한 역할을 할 것으로 생각된다.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2016년도 춘계학술대회 및 임시총회
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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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