• Title/Summary/Keyword: ketone

Search Result 660, Processing Time 0.023 seconds

Purification and Characterization of Oriental Pear(Niitaka, Pyrus pyrifolia Nak.) Protease (동양배(신고(新高)) Protease의 정제(精製) 및 성질(性質)에 관(關)하여)

  • Kim, Seung Yeol;Chung, Hai Jung;Kim, Seung Kyeom;Shin, Cheol Seung
    • Korean Journal of Agricultural Science
    • /
    • v.16 no.2
    • /
    • pp.225-238
    • /
    • 1989
  • These studies were conducted to investigate the extraction, purification and characterization of oriental pear (Niitaka. Pyrus pyrifolia Nak.) protease, and the results obtained were as follows: 1. Oriental pear protease was effectively extracted by the method of homogenizing pear pulp with 0.7 volume of 0.1M-sodium phosphate buffer, pH 6.5 containing 5mM-cysteine, 40mM-2-mercaptoethanol and 2mM-EDTA at 10,000 rpm for 5 min. 2. The protease was purified by ammonium sulfate fractionation, Sephadex G-100 filtration and DEAE-Sephadex A-50 column chromatography, and the purified enzyme gave a single protein band on polyacrylamide gel electrophoresis. 3. The specific activity of purified enzyme was 29.65 unit/mg protein and the yield was 7.22%. 4. The moecular weight of the protease was estimated to be about 51,000 by SDS-polyacrylamide gel electrophoresis, and the enzyme had Km value of 54.5 mg/ml for casein. 5. The purified enzyme had a maximum activity at pH 6.0 and $50^{\circ}C$, and was stable from pH 5.5-6.5 and at temperatures below $50^{\circ}C$ 6. Casein was a better substrate for this protease compared to hemoglobin. 7. The enzyme activity was markedly inhibited by p-chloromercuribenzoic acid and heavy metal salts such as $HgCl_2$ and $MnSO_4$ also considerably inhibited the enzyme activity.

  • PDF

Comparison of Passive Diffusional Organic Vapor Samplers with Charcoal Tubes for the Measurement of Mixed Organic Solvents (확산형 포집기와 활성탄관을 이용한 공기중 혼합 유기용제 측정에 관한 연구)

  • Ahn, Kyu-Dong;Yeon, You-Yong;Lee, Byung-Kook
    • Journal of Korean Society of Occupational and Environmental Hygiene
    • /
    • v.4 no.2
    • /
    • pp.127-136
    • /
    • 1994
  • Diffusional sampling devices offer many advantages for measuring concentration levels of industrial contaminants than the conventional pump and charcoal tubes because they are lightweight, require no power, pump or tubing. This study designed to evaluate and compare the sampling performance of passive sampler to charcoal tube from mixed organic solvent workplace with 181 organic solvent using workers working in different concentration of organic solvents. All study workers kept both devices in their breathing zone simultaneuosly in the workplaces, and the sampling analytical results were compared with those of charcoal tube. The results obtained are as follows: 1. The concentrations of toluene and xylene measured by passive sampler were slightly higher than those of charcoal tube, but there were no significant statistical differences between two methods. 2. The concentrations of MEK and cyclo-hexanone measured by passive sampler in low exposure workplace (below 0.20 of MEK TLV levels and 0.1 of cyclo-hexanone TLV levels) were about 2 times higher than that of charcoal tube sampling. While, absorption efficiency of passive sampler was reduced according to increasing concentration measurements of MEK and cyclo-hexanone in air. 3. The ratios of concentrations of toluene, xylene, MEK and cyc1o-hexanone measured by passive sampler over those measured by charcoal tube were 1.11, 1.07, 1.63 and 3.65 respectively. 4. The percentages of concentration of passive samplers within 0.75 and 1.25 of charcoal tube value as a reference value of 1.0 were 57% in toluene, 74% in xylene, 34% in MEK and 32% in cyclo-hexanone respectively. 5. The correlation coefficients of toluene, xylene, MEK and cyclo-hexanone between passive sampler and charcoal tube sampler were 0.963, 0.957, 0.943 and 0.562 with statistical significance.

  • PDF

Studies on the storage of fresh fruits and vegetables by plastic coating (1) on Rall's Janet Apple- (Plastic Coating에 의(依)한 청과물저장연구(靑果物貯藏硏究) (제1보(第1報)) -사과(국광(國光))에 대(對)하여-)

  • Park, Nou-Pung
    • Applied Biological Chemistry
    • /
    • v.12
    • /
    • pp.89-97
    • /
    • 1969
  • These studies were made on coating of fresh fruits and vegetables with PVC under view on preservation of fresh fruits and vegetables due to restrain water evaporation of them and control gas metabolisms. The results obtained, as selection of PVC materials availble for preservation of them, procedure of the coating, useful organic solvents, concentration of PVC solution and the time of dipping, and effects of the preservation of them, were summarized as follows: 1) PVC powder 222 and PVC powder 443 were surveyed as suitable materials for coating apple, and later was excellent in lustrous aspect. 2) The coating procedure which dipt into PVC solution was suitable to treat much within a short space of time, and using of ventillator accelerate rather evaporation of the organic solvent. 3) Aceton and methyethylketon as solvent of the PVC were, in purpose of storage only, avaible, while aceton was, in view ef taste, better. 4) 10% of the PVC solutions were better to preserve the freshness of apple and 5, 4, 1 and 0.5% of them in order were gradually decreased in preservability, and 15% of them as high concentration were looked like fermentation in one week after the coating. 5) The dipping time was also better n minute than 10 seconds in a preservation but 1-2 minutes could be applied owing to be not affected of the taste due to coating. 6) The freshness of treatment groups were extend about 48 days after coating in room temperature but control lost market value with 12 days. 7) Weight of control was decreased to 10-13% but treatment which was dipped into 10% of coating solution for one minute was decreased to 3.5-4.6% and treatment for four minutes was did to 2.9-3.0%. 8) Change in respiration was less is treatment groups than control in exhaustion of $CO_2$, and water soluble sugar, reduced sugar and pH were not changed almost due to coating. 9) Pannel discussion of the taste was indicated that control was better than treatments soon after coating, but treatments were rather than control last period of storage and treatment with aceton solvent specially was better than others.

  • PDF

The Role of Interleukin 8 and NF(nuclear factor)-κB in Rhinovirus-Induced Airway Inflammation (Rhinovirus 유발성 기도염증반응에서 Interleukin-8과 전사인자 NF(nuclear factor)-κB의 역할에 대한 연구)

  • Yoon, Ho Joo;Kim, Mi Ok;Sohn, Jang Won;Kim, Jung Mogg;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
    • /
    • v.54 no.1
    • /
    • pp.104-113
    • /
    • 2003
  • Background : Rhinovirus(RV) infections frequently trigger dyspnea and paroxysmal cough in adult patients with asthma and are the most prevalent cause of the common cold. However, the mechanisms of a RV-induced airway inflammation is unclear. Since the RV does not directly destroy the airway epithelium, it is presumed that the immune response to the RV contributes to the pathogenesis of the respiratory symptoms. In order to test this hypothesis, this study characterized the time-sequenced alterations in interleukin(IL)-8 elaboration from the human bronchial epithelial cells and evaluated the role of NF(nuclear factor)-${\kappa}B$ in the RV-induced IL-8 production by pretreating the inhibitors of NF-${\kappa}B$ activation. Methods : The ability of RV-infected human bronchial epithelial cells and BEAS-2B cells to produce the IL-8 was compared with the controls. This study infected BEAS-2B cells with the RV14 obtained from the American Type Culture Collection. The supernatants were harvested from the RV infected BEAS-2B cells and the controls at 2hr, 4hr, 6hr, 12hr, 24hr, 48hr from the inoculation time. This study measured the IL-8 concentration using the ELISA kits. In order to elucidate the role of NF-${\kappa}B$ in the RV-induced IL-8 production, the effect of the NF-${\kappa}B$ inhibitors was evaluated on RV-induced IL-8 production. Results: The BEAS-2B cells produced small amounts of IL-8 that accumulated slowly with time in the culture. The RV was a potent stimulator of the IL-8 proteins production by BEAS-2B human bronchial epithelial cells. Antioxidants, N-acetyl-L-cysteine(NAC),\ and pyrrolidine dithiocarbamate(PDTC), blocked the IL-8 elaboration by the RV-infected BEAS-2B cells, which was dose-dependent, but N-Tosyl-L-phenylalanine chloromethyl ketone(TPCK) did not. Conclusion: Some antioxidants inhibited the RV-induced IL-8 production by blocking the NF-${\kappa}B$, which may have a therapeutic potential in asthma.

Flavor Components in Mash of Takju Prepared by Different Raw Materials (원료를 달리하여 담금한 탁주 술덧의 향기성분)

  • Lee, Joo-Sun;Lee, Taik-Soo;Park, Sung-Oh;Noh, Bong-Soo
    • Korean Journal of Food Science and Technology
    • /
    • v.28 no.2
    • /
    • pp.316-323
    • /
    • 1996
  • Flavor components in mash of Takju prepared by different raw materials such as nonglutinous rice, glutinous rice, barley and wheat flour were detected by GC and GC-MS method using non-polar column. Seven alcohols, 15 esters, 10 organic acids, 1 aldehyde, 4 benzenes, 3 phenols, 8 alkans, 2 ketones and 5 others were found in takju after 16 day of fermentation. takju by wheat flour had the most various components of volatile flavor. Treatment with addition starter had less flavor component than that without addition starter in takju by nonglutinous rice. Nine kinds of flavor components including acetic acid ethyl ester, 3- methyl-1-butanol, acetic acid, ethyl benzene, acetic acid 3-methyl butyl ester, 2-phenylethanol, 2,6-di-tert-butyl-4-methyl phenol. plumbagic acid and 1,2-benzenedicarboxylic acid dibutyl ester were commonly detected in all the treatments. Especially, 2,4,0-trimethyl-1,3-benzenediamine was isolated in takju that was made of nonglutinous rice without addition starter. Diethyl sulfide, 4-methoxy benzaldehyde, docosane and 2-methyl propyl octadecanoic acid were isolated from takju by nonglutinous rice with addition starter. Propionic acid ethyl ester, acetic acid butyl ester, 2-methyl butane and 3-methyl pentane were isolated from takju glutinous rice. 2-Hydroxy-4-methyl pentanoic acid and 2-methyl tridecane were isolated from akju by barley 3-(Methylthio)-1-propanol. hexanoic acid ethyl ester, butanoic acid monomethyl ester, tridecanoic acid, ethyl tetramethyl cyclopentadiene and 1,5-diaza-2,9-diketocyclotetradecane were isolated from takju by wheat flour. Major volatile flavor components were acetic acid ethyl ester, 3-methyl-1-butanol, acetic acid and 2-phenylethanol.

  • PDF

Potential Antitumor $\alpha$-Methylene-$\gamma$-butyrolactone-Bearing Nucleic Acid Base. 3. Synthesis of $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans

  • Kim, Jack-C.;Kim, Si-Hwan;Kim, Ji-A;Choi, Soon-Kyu;Park, Won-Woo
    • Archives of Pharmacal Research
    • /
    • v.21 no.4
    • /
    • pp.458-464
    • /
    • 1998
  • Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

  • PDF

Study for the Synthesis of $[^{123}I]$Idoxifene and Its Uptake in the Breast Cancer Cell ($[^{123}I]$Idoxifene 합성과 유방암의 세포섭취에 관한 연구)

  • Cho, Young-Sub;Yang, Seung-Dae;Suh, Yong-Sup;Chun, Kwon-Soo;Ahn, Soon-Hyuk;Lim, Soo-Jung;Lim, Sang-Moo;Yu, Kook-Hyun
    • The Korean Journal of Nuclear Medicine
    • /
    • v.34 no.5
    • /
    • pp.410-417
    • /
    • 2000
  • Purpose: Idoxifene is currently entering phase II clinical trials for the treatment of advanced breast cancer. The radiolabeled idoxifene using $[^{123}I]$ provides an opportunity for clinical pharmacology with single photon emission computed tomography (SPECT). The purpose of this study was to prepare radiolabeled idoxifene using $[^{123}I]$ and to determine its cell uptake of breast cancer cell line. Materials and Methods: With a view to evaluating new anticancer drugs, we are investigating the novel antiestrogen pyrrolidino-4-iodotamoxifen (idoxifene). $[^{123}I]$Idoxifene has been prepared in no-carrier-added form using a tributyl stannylated precursor which has been synthesized by means of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid on the basis of previously reported standard methods. The biodistribution and dynamic behavior of the compound were investigated using the comparative breast cancer cell line, MCF-7 (estrogen receptor-positive) and MDA-MB-468 (non-estrogen receptor). Results and Conclusion: Acylation of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid gave the versatile ketone (81%) which reacted with 1,4-diiodobenzene to give triphenylethylene as a mixture of E and Z geometric isomers, which were separated by the recrystallization in ethanol. The E-isomer was treated with pyrrolidine to give idoxifene (67%). In order to incorporate radioactive iodine into the 4-position, the 4-stannylated precursor was prepared (30%). The yield of radioiodination was 90-92% with a high radiochemical purity greater than 98%. The ratio of tumor uptake of the breast cancer cell line between MCF-7 and MDA-MB-468 was about 1.7.

  • PDF

Development of VOCs Treatment Technology using High Efficiency Hybrid System with Multi-Scrone (멀티 선회류식 세정장치를 이용한 고효율 하이브리드 VOCs 습식처리 SYSTEM 개발)

  • Lim, Seong-Il;Kim, Nor-Jung;Kim, Sun-Mi;Lee, Seong-Hun;Kim, Sun-Uk;Chang, Won-Seok;Park, Dae-Won;Kim, Lae-Hyun;Kim, Jae-Hyung
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.31 no.7
    • /
    • pp.491-498
    • /
    • 2009
  • We studied to develop high-efficiency removal system of odor and VOCs(Volatile Organic Compounds) from environmental infrastructure facilities and oil refineries, painting facilities and so on. It can replace RTO and RCO. We tried an removal experiment for VOCs (toluene, xylene, benzene, MEK(methyl ethyl ketone), ethanol, formalin etc. and odor compounds (hydrogen sulfide, etc.). In process, as pre-treatment we used the scrubber with vortex flow (Multi-scrone) to remove the hydrophilic VOCs and as post-treatment, used fibrous bio-filter to remove the hydrophobic VOCs. This hybrid system remove with high efficiency both the hydrophilic VOCs and hydrophobic VOCs. And we tried to make this system to be compact. In experiment using Multi-scrone, contact time is 2~3 seconds and absorption scrubbing water is diaphragm-type electrolysis water. hydrophilic VOCs like ethanol and relatively hydrophilic odor compounds like hydrogen sulfide is excellent, these substances has been removed almost completely, respectively 95~99%, 93~97%. And for MEK, formalin also Showed a high removal efficiency, respectively 78~90%, 72~85%. But in experiment using Multi-scrone, the hydrophobic VOCs like BTX showed a low removal efficiency, respectively 16~22%, 12~18%, 8~16%. In hydrophobic VOCs, toluene removal experiment using fibrous bio-filter, early efficiency was low but after 10days, adaptation period showed high efficiency 85~95%. but in the mixed phase, toluene and MEK efficiency reduced 5~10%. this show microorganism treat first MEK easy to remove. The removal efficiency for MEK using the fibrous biofilter was stable, 80~92%. This hybrid system is also high economical efficiency for RTO. This system reduce more than 50% the cost of equipment and maintenance. As a result, we expect this technology is in the limelight as high efficiency treatment of VOCs in mid-low price.

Quality Properties of White Lotus Leaf Fermented by Mycelial Paecilomyces japonica (동충하초 균사체로 발효시킨 백련잎차의 품질특성)

  • Kim, Jong-Suk;Wang, Su-Bin;Kang, Seong-Koo;Cho, Young-Sook;Park, Seok-Kyu
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.38 no.5
    • /
    • pp.594-600
    • /
    • 2009
  • Quality characteristics of white lotus leaf tea (LLT) fermented with or without mycelial Paecilomyces japonica were investigated. Extraction yield and browning index of hot water extract from non fermented and fermented LLTs were higher than those of ethanol extract (p<0.05). In all LLTs, nutritional components such as total free sugar, free amino acids and minerals of hot water extracts were higher than those of ethanol extracts except for total organic acids (p<0.05). Contents of total free sugar and organic acids were markedly increased through fermentation process of mycelial Paecilomyces japonica. in the same solvent extracts (p<0.05). Contents of most taste components of fermented LLT were increased by mycelial solid fermentation (p<0.05), but total free amino acids of two extracts were decreased in the range of $37.1{\sim}67.2%$ as compared to non-fermented LLT. Fifty-nine volatile compounds were identified by GC and GC-MS, including 11 aldehydes, 14 alcohols, 11 ketones, 11 hydrocarbones and 12 acids. Aldehyde and ketone compounds were more identified in fermented LLT than in non-fermented LLT being abundant alcohol compounds by simultaneous steam distillation and extraction. The most abundant compounds of LLT identified in this study were curcumene followed by 2,6-bis(1,1-dimethylethyl)-4-methyl-phenol and cyclohexen. Main compounds of fermented LLT were 2,6-bis(1,1-dimethylethyl)-4-methyl-phenol, butanoic acid, furfural, benzaldehyde, hexanoic acid and 2(3H)-furanone.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
    • /
    • 2016.05a
    • /
    • pp.19-20
    • /
    • 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.

  • PDF