著者

犀川 陽子 岡本 博樹 乾 泰地 橋本 貴美子 中田 雅也 真壁 みどり 奥野 智旦 須田 隆

出版者

天然有機化合物討論会実行委員会

雑誌

天然有機化合物討論会講演要旨集

巻号頁・発行日

vol.43, pp.443-448, 2001

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A poisonous mushroom Podostoma cormu-damae caused two lethal poisoning in Japan in 1999 and 2000. Some of the following symptoms are observed in these poisonings: gastrointestinal disorder, erroneous perception, decrease in the number of leukocytes and thrombocytes, deciduous skin of face, loss of hair, and atrophy of the cerebellum which brings about a speech impediment and voluntary movement problems. We studied the toxic constituents of its culture broth and the fruit bodies using lethal effect on mice as an index. The extracts from the culture filtrate and the fruit bodies were injected into the abdominal cavity of a mouse. The lethal effect was observed in both extracts from the culture filtrate and the fruit bodies. The organic extracts of the culture filtrate were chromatographed on silica gel to give the major compounds 1, 2, and 3. The ^1H and ^<13>C NMR spectroscopic analyses revealed that these compounds 1〜3 are members of the macrocyclic trichothecene group. Comparison of the spectral data of 1〜3 with those in the literature revealed that 1 is roridin E, 2 is verrucarin J (muconomycin B), and 3 is satratoxin H. On the other hand, the fruit bodies were extracted with water and methanol. The water extracts were chromatographed on ODS to give satratoxin H (3), and the methanol extracts were chromatographed on silica gel to give 4〜6. The NMR and MS analyses showed that 4, 5, and 6 is the 12',13'-diacetate, 12'-acetate, and 13'-acetate of satratoxin H (3), respectively. The 4〜6 are new compounds that occur in nature. All these macrocyclic trichothecenes except for 2 had a lethal effect on mice by at least 0.5 mg per capita.

著者

野副 重男 小池 隆 辻 恵美 草野 源次郎 瀬戸 治男 青柳 富貴子 松本 春樹 松本 毅 奥野 智旦

出版者

天然有機化合物討論会

雑誌

天然有機化合物討論会講演要旨集

巻号頁・発行日

no.25, pp.282-289, 1982-09-10

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In the course of our continuing investigation of the constituents of a hallucinogenic mushroom, Gymnopilus spectabilis (Japanese name, O-waraitake) collected at Oguni, Yamagata, we have isolated novel type of polyisoprenepolyols designated as gymnopilins -A, -B, and gymnoprenols -A, -B, -C, and -D etc. These substances occured as a mixture of some homologues were separated and throughly purified by means of preparative HPLC and their chemical structures were determined on the basis of the results of oxidative degradation as well as their spectroscopic properties. These polyisoprenepolyols were shown to contain same repeated unit corresponding to hydrated form of usual isoprenoid chain, two or three double bonds and a terminal vicinal diol moiety or the corresponding ester with 3-hydroxy-3-methyl-glutaric acid. The location of double bond was confirmed by the degradation studies. The structures of polyisoprenepolyols such as 1a and 2a have never been encountered in natural products obtained so far. The stereochemistry and biological activities are under investigation.

著者

田中 正泰 針ヶ谷 弘子 鎌田 樹志 氏原 一哉 橋本 勝 橋本 貴美子 松田 冬彦 柳屋 光俊 白濱 晴久 奥野 智旦

出版者

天然有機化合物討論会実行委員会

雑誌

天然有機化合物討論会講演要旨集 34 (ISSN:24331856)

巻号頁・発行日

pp.110-117, 1992-09-10 (Released:2017-08-18)

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Ohwaraitake is Japanese name of a poisonous mushroom Gymnopilus spectabilis and means "a loud laugh mushroom". Accidental ingestion of it causes hallucinosis and abnormal behavior. A hallucinogenic mushroom contains usually psilocybin or its analogues but it is reported that any psilocybin or its congenor does not found in Japanese Ohwaraitake. The toxic symptoms suggest that any neuroexcitatory substance must be contained. The isolation was carried out monitoring depolarizing activity on the new born rat spinal cord. 1. Chromatographical fractionation and bisassay showed that the neuroexcitatory active compounds were gimnopilins (G) which were known as bitter principles of this fungus. Further fractionation revealed that G (1) was inactive and activities of G (2) increased from n=7 to n=5 and besides, G (2), which was newly isolated this time, was more active than G (2). 2. The structure of the new G was determined by the chemical degradation as formula 3. 3. The Chirality of hydroxymethylglutamic acid (HMGA) part was determined as S configuration by the acquisition of (R)-mevalonaloctone through LiBH4 reduction or G (1 and 2). 4. G (2) does not work as an ion carrier probably, since it does not particularly take metal ion from aqueous solution into an organic solvent. 5. Determination of the chirality of tert-alcohols in gymnoprenol (G'), that is, G without HMGA part, was attempted. Eight isomers of the model compounds corresponding to G' (m=2, n=4) were synthesized and were not discriminated by HPLC and NMR. Since two diastereoisomers of MTP-ethoxy-methoxy derivatives of G' (m=1, n=2) were distinguished by ^1H and ^<19>FNMR, these derivatives may be applicable to identification of the synthetic and the natural G'.

著者

清水 哲哉 中津 亨 宮入 一夫 奥野 智旦 加藤 博章

出版者

日本応用糖質科学会

雑誌

Journal of Applied Glycoscience (ISSN:13447882)

巻号頁・発行日

vol.51, no.2, pp.161-167, 2004 (Released:2008-03-24)

参考文献数

28

被引用文献数

1 2

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エンドポリガラクツロナーゼ(EndoPG)は,inverting型の加水分解酵素であり,その触媒機構の詳細は不明であった。本研究では,リンゴ銀葉病菌(Stereum purpureum)の病徴発現蛋白として単離されたEndoPG Iを用いて触媒機構の解明を目的としてX線結晶構造解析を行った。イオンスプレー型質量分析装置を用いたEndoPG Iの糖鎖修飾の分析結果からEndoPG Iの結晶化には,3種のアイソフォーム,EndoPG Ia,Ib,Icのうち糖鎖の最も少ないEndoPG IaをEndoHで糖鎖切断した脱糖鎖型EndoPG Iaを用いた。この脱糖鎖型EndoPG IaをPEG 4000を沈殿剤に用いたハンギングドロップ蒸気拡散法により結晶化した。得られた結晶は,空間群P1に属し,格子定数はa=37.26 Å,b=46.34 Å,c=52.05 Å,α=67.17°,β=72.44°,γ=68.90°であった。この結晶を用いて大型放射光施設SPring-8によりX線回折実験を行った。結晶構造の決定は,多波長異常分散法で行った。SPring-8で収集したデータを用いて初期位相を決定し,最終的に0.96 Å分解能でR値11.4%,Rfree値14.0%という超高分解能のEndoPG Iの構造モデルを得た。得られたEndoPG Iの構造は,10回の完全ターンの平行β-へリックス構造であった。さらに反応生成物であるガラクツロン酸(GalA)を1分子を含むbinary複合体と2分子含むternary複合体の結晶構造を1.00 Åおよび1.15 Å分解能で決定した。binary複合体において1分子のピラノース型(GalpA)の結合がみられた。GalpAの結合位置は,活性残基と推定されているAsp153,Asp173,Asp174の還元末端側にあたることから,GalpAは+1サブサイトに結合していると考えられた。一方,ternary複合体においては,GalpAの他に,フラノース型のGalfAの結合がみられた。GalfAの結合位置は,活性残基の非還元末端側にあたる-1サブサイトと決定された。+1サブサイトでは,三つの塩基性残基,His195,Arg226,Lys228がGalpAのカルボン酸の結合に関与していた。一方,-1サブサイトでは,特異なcis型のペプチド結合がGalfAのカルボン酸の認識に関わっていた。そこで,二つのガラクツロン酸の結合状態を基に,基質の-1,+1結合した2量体部分のモデルを構築し,EndoPG Iの反応機構を検討した。まず,Asp173は切断される基質モデルのグリコシド結合と直接水素結合しうる位置にあった。したがって,Asp173はグリコシド結合にプロトンを供給する一般酸触媒残基と確認された。一方,Asp153とAsp174は,基質のアノマー炭素を求核攻撃すると考えられる水分子と水素結合していた。このことから,Asp153あるいはAsp174が水からプロトンを引き抜き活性化する一般塩基触媒残基と予想された。

著者

田中 正泰 針ヶ谷 弘子 鎌田 樹志 氏原 一哉 橋本 勝 橋本 貴美子 松田 冬彦 柳屋 光俊 白濱 晴久 奥野 智旦

出版者

天然有機化合物討論会

雑誌

天然有機化合物討論会講演要旨集

巻号頁・発行日

no.34, pp.110-117, 1992-09-10

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Ohwaraitake is Japanese name of a poisonous mushroom Gymnopilus spectabilis and means "a loud laugh mushroom". Accidental ingestion of it causes hallucinosis and abnormal behavior. A hallucinogenic mushroom contains usually psilocybin or its analogues but it is reported that any psilocybin or its congenor does not found in Japanese Ohwaraitake. The toxic symptoms suggest that any neuroexcitatory substance must be contained. The isolation was carried out monitoring depolarizing activity on the new born rat spinal cord. 1. Chromatographical fractionation and bisassay showed that the neuroexcitatory active compounds were gimnopilins (G) which were known as bitter principles of this fungus. Further fractionation revealed that G (1) was inactive and activities of G (2) increased from n=7 to n=5 and besides, G (2), which was newly isolated this time, was more active than G (2). 2. The structure of the new G was determined by the chemical degradation as formula 3. 3. The Chirality of hydroxymethylglutamic acid (HMGA) part was determined as S configuration by the acquisition of (R)-mevalonaloctone through LiBH4 reduction or G (1 and 2). 4. G (2) does not work as an ion carrier probably, since it does not particularly take metal ion from aqueous solution into an organic solvent. 5. Determination of the chirality of tert-alcohols in gymnoprenol (G'), that is, G without HMGA part, was attempted. Eight isomers of the model compounds corresponding to G' (m=2, n=4) were synthesized and were not discriminated by HPLC and NMR. Since two diastereoisomers of MTP-ethoxy-methoxy derivatives of G' (m=1, n=2) were distinguished by ^1H and ^<19>FNMR, these derivatives may be applicable to identification of the synthetic and the natural G'.

著者

山崎 賀久 徳永 隆司 奥野 智旦

出版者

社団法人 日本食品科学工学会

雑誌

日本食品科学工学会誌 (ISSN:1341027X)

巻号頁・発行日

vol.52, no.4, pp.160-166, 2005 (Released:2007-04-13)

参考文献数

21

被引用文献数

6 19

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強陽イオン交換 (SCX) カラムを使用し, 10mmol/Lリン酸二水素カリウム (pH2.50) を移動相とし, 誘導化なしに紫外吸光検出する高速液体クロマトグラフィ (HPLC) を用いたニンニク中の11種のフレーバ前駆体 (S-アルキルシステイン誘導体), S-methyl cysteine S-oxide (methiin), (+)-S-(2-propenyl) cysteine S-oxide (alliin), (-)-S-(2-propenyl) cysteine S-oxide (allo alliin), S-(E -1-propenyl) cysteine S-oxide (isoalliin), 5-methylthiomorpholine-3-carboxylic acid S-oxide (cycloalliin), S-(2-propenyl) cysteine (deoxyalliin), N-(γ-glutamyl)-S-methylcysteine, N-(γ-glutamyl)-S-(2-propenyl) cysteine, N-(γ-glutamyl)-S-(E -1-propenyl) cysteine, N-(γ-glutamyl)-S-(2-propenyl) cysteine S-oxide及びN-(γ-glutamyl)-S-(E -1-propenyl) cysteine S-oxideの定量分析法を新たに確立した.本分析法を11種類の試料に適用したところ, 報告例の少ないisoalliin, cycloalliinのニンニク中含有量は, 11種類の平均値でそれぞれ0.13%と0.16% (乾物換算) であった. また, 天然物としての報告がないallo alliin及び天然物としては未確認のN-(γ-glutamyl)-S-2-propenyl cysteine S-oxideが全試料で天然物として検出され, その平均含有量はそれぞれ0.06%と0.16% (乾物換算) であった.