66 4-2 ᐇ㦂᪉ἲ

66

67

4-2-2 ᕷ㈍ࢱࣥࢼ࣮ࢮࡢάᛶ ᐃ

HPLCศᯒࢆ⏝࠸࡚㸪ᚓࡽࢀࡓࢡ࣐ࣟࢺࢢ࣒ࣛࡢ gallic acid ࡢࣆ࣮ࢡ࠿ࡽάᛶࢆホ౯ ࡋࡓࠋ

ࡲࡎ㸪ᇶ㉁࡛࠶ࡿࢱࣥࢽࣥ㓟࡜ࢱࣥࢼ࣮ࢮࢆ⁐ゎࡍࡿ ࡓࡵ࡟౑⏝ࡍࡿ0.05 mol / l ࢡ

࢚ࣥ㓟⦆⾪ᾮ㸦pH 5.5㸧ࢆㄪ〇ࡋࡓࠋ0.05 mol / l ࢡ࢚ࣥ㓟⦆⾪ᾮ㸦pH 5.5㸧ࡣ㸪0.05 mol/l

ࡢࢡ࢚ࣥ㓟 2Ỉ⣲࣒࢝ࣜ࢘Ỉ⁐ᾮ࡜ 0.05 mol / l NaOHỈ⁐ᾮࢆΰྜࡋ㸪pH 5.5࡟ㄪᩚ

ࡋࡓࠋ

ᇶ㉁࡛࠶ࡿࢱࣥࢽࣥ㓟ࡣ0.32 %㸪㓝⣲࡛࠶ࡿࢱࣥࢼ࣮ࢮࡣ 3 mg / 100 ml࡜࡞ࡿࡼ ࠺

࡟ 0.05 mol / l ࢡ࢚ࣥ㓟⦆⾪ᾮ㸦pH 5.5㸧࡟⁐ゎࡋࡓࠋࢱࣥࢽࣥ㓟⁐ᾮ 500ȣlࢆ࢔ࣝ

࣑ࣈࣟࢵࢡࢆ⏝࠸࡚ 10 ศ㛫㸪30Υ࡛ணഛຍ⇕ࡋࡓᚋ㸪100ȣlࢆࢧࣥࣉࣜࣥࢢࡋ࡚཯

ᛂ๓ࢧࣥࣉࣝ࡜ࡋࡓࠋṧࡗࡓᇶ㉁⁐ᾮ࡟ࡣࢱࣥࢼ࣮ࢮ⁐ᾮ 100ȣlࢆຍ࠼࡚࣎ࣝࢸࢵࢡ

ࢫ࣑࢟ࢧ࣮࡛ᨩᢾᚋ㸪30Υࡢ࢔࣑ࣝࣈࣟࢵࢡ୍࡛᫬㛫ຍ ࡋࡓࠋ཯ᛂ⤊஢ᚋ㸪100ȣl

ࢆࢧࣥࣉࣜࣥࢢࡋ࡚཯ᛂᚋࢧࣥࣉࣝ࡜ࡋࡓࠋ཯ᛂ๓ࢧࣥࣉࣝ㸪཯ᛂᚋࢧࣥࣉࣝඹ࡟ 100 ȣlࡢ㓑㓟࢚ࢳ࡛ࣝᢳฟࡋ㸪㓑㓟࢚ࢳࣝᒙࡢ༙㔞㸦50ȣl㸧ࢆ஝⇱ᚋ㸪࣓ࢱࣀ࣮ࣝ 50ȣ l࡟⁐ゎࡋ࡚࠺ࡕ 20ȣlࢆHPLC ศᯒ࡟౪ࡋࡓࠋ

ḟ࡟㸪ࢱࣥࢼ࣮ࢮάᛶࢆ⤒᫬ⓗ࡟ほᐹࡍࡿᐇ㦂ࢆ⾜ࡗࡓࠋ4 mlࡢࢱࣥࢽࣥ㓟⁐ᾮ࡟

1 mlࡢࢱࣥࢼ࣮ࢮ⁐ᾮࢆΰྜࡋ㸪཯ᛂ㛤ጞ࠿ࡽ 5 ศ࠾ࡁ࡟ 50ศ㛫㸪200ȣlࡢ཯ᛂᾮ

ࢆࢧࣥࣉࣜࣥࢢࡋࡓࠋࡑࢀࡒࢀ 200ȣlࡢ㓑㓟࢚ࢳ࡛ࣝᢳฟᚋ㸪100ȣlࡢ㓑㓟࢚ࢳࣝᒙ

ࢆᅇ཰ࡋ㸪ࡇࢀࢆ஝⇱ࡉࡏ࡚࠿ࡽ࣓ࢱࣀ࣮ࣝ 50ȣl࡟⁐ゎࡋ࡚࠺ࡕ 20ȣlࢆHPLCศ ᯒ࡟౪ࡋࡓࠋ

68

69

4-2-3 ೺඲࣑ࢬࢼࣛ㎶ᮦᢳฟ≀ࡢᾮᾮศ㓄

➨୍❶࡛♧ࡋࡓ㏻ࡾ㸪೺඲࣑ࢬࢼࣛᮦࡣ 2006ᖺ 11᭶࡟ᒱ㜧኱Ꮫ₇⩦ᯘࡼࡾධᡭࡋࡓࠋ

㎶ᮦࡢᢳฟ≀࡜ࡋ࡚ NS-eࢆᚓ࡚㸦➨୍❶ཧ↷㸧㸪ࡇࢀࢆỈ࡟ᠱ⃮ࡋ㸪࢚࣮ࢸࣝ㸦3L™ 3 ᅇ㸧㸪㓑㓟࢚ࢳࣝ㸦3L™8 ᅇ㸧㸪n-ࣈࢱࣀ࣮ࣝ㸦3L™3ᅇ㸧࡟ࡼࡗ࡚ᾮᾮศ㓄ࡋࡓࠋᚓ

ࡽࢀࡓྛྍ⁐㒊ࡣ࢚ࣂ࣏࣮ࣞࢱ࣮ࢆ⏝࠸࡚⃰⦰஝ᅛࡋ཰㔞ࢆồࡵ㸪ࡲࡓࡑࡢ୍㒊ࡣ࣓

ࢱࣀ࣮ࣝ࡟෌ᗘ⁐ゎࡋ࡚ HPLC ศᯒ࡟౪ࡋࡓࠋ

70

4-2-4 NS-EtOAc-Sࡢᕷ㈍ࢱࣥࢼ࣮ࢮฎ⌮࡜ HPLCศᯒ

࣑ࢬࢼࣛ㎶ᮦ 70 %࢔ࢭࢺࣥᢳฟ≀ࡢ㓑㓟࢚ࢳࣝྍ⁐㒊ࢆࢡ࢚ࣥ㓟 buffer㸦pH 5.5㸧࡟

⁐ゎࡋ࡚ࠊ0.32 %࡟ㄪ〇ࡋࠊᇶ㉁⁐ᾮ࡜ࡋࡓࠋࡇࡢᇶ㉁⁐ᾮ 2 ml࡟ࢱࣥࢼ࣮ࢮ⁐ᾮࢆ

ຍ࠼࡚཯ᛂ㛤ጞ࡜ࡋࡓࠋࢱࣥࢼ࣮ࢮ 0.3 mg / 10 mlࢆ500 μlຍ࠼ࡿ๓࡜ࠊࢱࣥࢼ࣮ࢮ

⁐ᾮࢆຍ࠼࡚ over night࡛཯ᛂࡉࡏࡓᚋࠊ཯ᛂᾮ 100 μlࢆࡑࢀࡒࢀᅇ཰ࡋࡓࠋ཯ᛂ๓

ᚋࡢࢧࣥࣉࣝ࡟ࡘ࠸࡚ࡑࢀࡒࢀ 200 μlࡢ㓑㓟࢚ࢳ࡛ࣝᢳฟࢆ⾜࠸ࠊ㓑㓟࢚ࢳࣝᒙࢆ

100 μlᅇ཰ࡋࡓࠋࡑࡋ࡚㓑㓟࢚ࢳࣝ␃ཤᚋࠊ50 μlࡢ࣓ࢱࣀ࣮ࣝ࡟෌ᗘ⁐ゎࡋ࡚ࠊࡑࡢ

࠺ࡕ 20 μlࢆ HPLCศᯒ࡟౪ࡋࡓࠋ

71

4-2-5 NS-EtOAc-Sῧຍᇵᆅ࡟࠾ࡅࡿࢼࣛ⳦ࡢᇵ㣴࡜㸪ᇵᆅᡂศࡢ HPLCศᯒ

Ț⥺⁛⳦῭ࡳࢩ࣮ࣕࣞ࡟ 㸪࣮࢜ࢺࢡ࣮ࣞࣈ࡟࡚⁛⳦㸦121Υ㸪15min㸧ࡋࡓ PDAᇵᆅ

ࢆὶࡋࡇࡳ㸪෭༷ࡋ࡚ᅛࡵࡓࠋࡇࡇ࡟㸪஦๓࡟ูࡢ PGAᇵᆅୖ࡟ᇵ㣴ࡋ෭ⶶᗜ࡛ಖ

⟶ࡋ࡚࠸ࡓࢼࣛ⳦ࢆ᳜⳦ࡋࡓࠋࡇࢀࢆ 28Υ࡛ 10 ᪥ᇵ㣴ࡋ㸪๓ᇵ㣴࡜ࡋࡓࠋ

50 ml ᐜ୕ゅࣇࣛࢫࢥ࡟ࣅࢱ࣑ࣥ࡜࣑ࢿࣛࣝࢆྵࡴᇵᆅ¹⁾ࢆ10 ml ㄪ〇ࡋ㸪⁛⳦ᚋ㸪࠶

ࡽ࠿ࡌࡵ๓ᇵ㣴ࡋ࡚࠾࠸ࡓ R. quercivora ࢆ๓ᇵ㣴ᇵᆅࡈ࡜ No. 4 ࡢࢥࣝࢡ࣮࣮࡛࣎ࣛ

ᡴࡕᢤ࠸࡚᥋✀ࡋࡓࠋࡑࡇ࡬ 㸪200 mg / ml 50 % ࣓ࢱࣀ࣮ࣝ⁐ᾮ࡟ㄪ〇ࡋࡓ

NS-EtOAc-Sࢆ20 μlῧຍࡋࡓࠋࣇࣛࢫࢥ࠶ࡓࡾࡢ NS-EtOAc-S⃰ᗘࡣ 4 mg / 10 ml ᇵ ᆅ ࡛࠶ࡗࡓࠋ28 Υ࡟ಖࡗࡓᇵ㣴ᗜ࡛ᇵ 㣴ࡋ㸪ᇵ㣴6 ᪥ᚋ࡟ᇵᆅ 1000 μlࢆࢧࣥࣉࣜ

ࣥࢢࡋࡓࠋࢧࣥࣉࣜࣥࢢࡋࡓᇵᆅ࡟ࡣከ㔞ࡢ⣒≧ࡢ⳦యࡀྵࡲࢀ࡚࠸ࡓࡢ࡛ 12,500

rpm, 5 min ࡢ᮲௳࡛㐲ᚰศ㞳ࡋ㸪ୖΎ500μlࢆᚓࡓࠋࡇࡇ࡟㓑㓟࢚ࢳࣝ 500μlࢆຍ࠼ΰ

ྜࡋ㸪㓑㓟࢚ࢳࣝᒙࢆ 300μlᅇ཰ࡋࡓࠋࡇࡢ㓑㓟࢚ࢳࣝ⁐ᾮ࡟❅⣲࢞ࢫࢆ྿ࡁ௜ࡅࡿ

ࡇ࡜࡛᏶඲࡟㓑㓟࢚ࢳࣝࢆ㝖ཤࡋ 㸪࣓ࢱࣀ࣮ࣝ 50μl࡟࡚⁐ゎࡋࡓࠋࡇࡢ࣓ࢱࣀ࣮ࣝ⁐

ᾮࡢ࠺ࡕ 20μlࢆ HPLCศᯒ࡟౪ࡋࡓࠋHPLCศᯒࡢ᮲௳ࡣ㸪3-2-1 ࡜ྠᵝ࡛࠶ࡗࡓࠋ

ࡲࡓ㸪ᇵ㣴๓ᚋ࡟⳦యࡢ୍㒊ࢆᅇ཰ࡋ㸪ගᏛ㢧ᚤ㙾(™100 or 200)࡟࡚ほᐹࡋࡓࠋ

72

4-3 ⤖ᯝ࡜⪃ᐹ

4-3-1 ᕷ㈍ࢱࣥࢼ࣮ࢮࡢάᛶ ᐃ

㉎ධࡋࡓ Aspergillus oryzae⏤᮶ࡢࢱࣥࢼ࣮ࢮ࡟ࡘ࠸࡚ࠊྠᑒࡉࢀ࡚࠸ࡓ㈨ᩱ࡟ࡣࡑࡢ

άᛶࡀ㺀30 units / mg௨ୖ㺁࡛࠶ࡿ࡜グ㍕ࡉࢀ࡚࠸ࡓࠋࡑࡇ࡛ࠊᐇ㝿࡟ࡇࡢࢱࣥࢼ࣮ࢮ ࡢάᛶࢆ▱ࡿࡓࡵ࡟ࠊ㓝⣲άᛶࡢ ᐃࢆ⾜ࡗࡓࠋ㉎ධࡋࡓࢱࣥࢼ࣮ࢮ࡟ྠᑒࡉࢀ࡚࠸

ࡓ㈨ᩱ࡟ᇶ࡙࠸࡚ᐇ㦂ࢆ⾜ࡗࡓ࡜ࡇࢁࠊ್ࡀ኱ࡁࡃࡪࢀࡓ(Fig. 4-1)ࠋࡑࡢࡓࡵࠊ㓝⣲

άᛶࡢィ⟬࡟ࡣ⮳ࡽ࡞࠿ࡗࡓࠋࡇࡢཎᅉ࡜ࡋ࡚ࠊ⣙ 1 ᖺ࡟ࢃࡓࡗ࡚෭ⶶᗜෆ࡛ࢱࣥࢼ

࣮ࢮࢆಖ⟶ࡋ࡚࠸ࡓࡓࡵ࡟ ࢱࣥࢼ࣮ࢮ⮬㌟ࡢάᛶࡀⴠࡕ࡚࠸ࡿྍ⬟ᛶ࡜ࠊᚤ㔞ࢆᢅ࠺

ࡓࡵ࡟ㄗᕪࡀ࡛࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ

ࡲࡓࠊ㈨ᩱ࡟グࡉࢀ࡚࠸ࡓࢱࣥࢼ࣮ࢮάᛶࡢ༢఩ࡢᐃ⩏࡟ࡣࠊ㺀pH 5.5ࠊ30Υ࡛ 1 ศ㛫

࡟ 1ȣmolࡢࢱࣥࢽࣥ㓟ࡢ࢚ࢫࢸࣝ⤖ྜࢆຍỈศゎࡍࡿ㓝⣲㔞ࢆ 1 unit࡜ࡍ ࡿ㺁࡜࠶ࡿ

ࡀࠊࢱࣥࢽࣥ㓟ࡣ」ᩘࡢ࢞ࣟࢱࣥࢽࣥ࠿ࡽ࡞ࡿΰྜ≀࡛࠶ࡿࡓࡵࠊࢱࣥࢽࣥ㓟 1ȣmol

࡜࠸࠺㔞ࢆ࡝ࡢࡼ࠺࡟ ᐃࡍࡿࡢ࠿࡜࠸࠺Ⅼ࡟␲ၥࡀ࠶ࡗࡓࠋࡑࡇ࡛ࠊpH 5.5ࠊ30Υ

࡛ 1ศ㛫࡟ຍỈศゎ࡟ࡼࡗ࡚ 1ȣmolࡢ࢞ࣜࢵࢡ㓟ࢆ⏕ᡂࡍࡿ㓝⣲㔞ࢆ 1 unit࡜ࡋ࡚ࠊ HPLCศᯒࡢࣆ࣮ࢡ㠃✚࠿ࡽ࢞ࣜࢵࢡ㓟ࡢ⏕ᡂ㔞ࢆィ⟬ࡋࠊάᛶࡢホ౯࡜ࡋ࡚⏝࠸ࡿ

஦ࡀ࡛ࡁࡿ࠿ࠊ᳨ウࡋࡓࠋ

ࢱࣥࢼ࣮ࢮ࡟ࡼࡿฎ⌮๓࡜ࠊ60ศࡢฎ⌮ᚋࠊࡑࢀࡒࢀࡢ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛࢆẚ㍑

ࡍࡿ࡜(Fig. 4-2)ࠊฎ⌮ᚋࡢ HPLCࢡ࣐ࣟࢺࢢ࣒࡛ࣛࡣ R.T. 26 min.௜㏆࡟ぢࡽࢀࡿ࢞ࣟ

ࢱࣥࢽࣥ┦ᙜࡢࣆ࣮ࢡࡀ㢧ⴭ࡟ῶᑡࡋࠊ㏫࡟࢞ࣜࢵࢡ㓟┦ᙜࡢࣆ࣮ࢡࡀቑຍࡋ࡚࠸ࡿ

ࡇ࡜ࡀࢃ࠿ࡗࡓࠋࡇࡢ஦࠿ࡽࠊHPLCศᯒ࡟ࡼࡗ࡚ࢱࣥࢼ࣮ࢮάᛶࢆホ౯ࡍࡿࡇ࡜ࡀ

73

ྍ⬟࡛࠶ࡿ࡜⪃࠼ࡓࠋ

ࡑࡇ࡛ࠊࢱࣥࢼ࣮ࢮ࡟ࡼࡿฎ⌮ᚋ 5ศࠊ10 ศ͐࡜⤒᫬ኚ໬ࢆ㏣ࡗࡓ࡜ࡇࢁࠊ࢞ࣜࢵࢡ 㓟ࡢ⏕ᡂ㔞ࡣ᫬㛫ࡢ⤒㐣࡟కࡗ࡚ቑຍࡋࡓ(Fig. 4-3)ࠋ

ࡲࡓࠊ࢞ࣜࢵࢡ㓟㔞࡟ࡘ࠸࡚ࡣ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛࡢࣆ࣮ࢡ㠃✚࡜⃰ᗘࡢ㛵ಀ࠿ࡽ

᳨㔞⥺ࢆసᡂࡋ(Fig. 4-4)ࠊࡇࢀࢆඖ࡟࢞ࣜࢵࢡ㓟㔞ࢆィ⟬ࡋࡓࠋ

Fig. 4-3ࡢ㏆᫬᭤⥺ࡢᘧࡼࡾ 1ศ㛫࡟⏕ᡂࡋࡓ࢞ࣜࢵࢡ㓟㔞ࡣ 6.1™10^-4 μmol࡜ィ⟬࡛

ࡁࡓࠋࡇࡢ㔞ࡣ HPLC ศᯒ࡟౪ࡋࡓ 20ȣlࡢ࣓ࢱࣀ࣮ࣝ⁐ᾮ୰ࡢ㔞࡛࠶ࡾࠊ཯ᛂᾮࡣ

඲య㔞࡟᥮⟬ࡋࡓ࡜ࡇࢁࠊ཯ᛂᾮ඲య࡛ࡢ 1ศ㛫ࡢ࢞ࣜࢵࢡ㓟⏕ᡂ㔞ࡣ

࡛࠶ࡗࡓࠋࡇࡢ࢞ࣜࢵࢡ㓟ࢆ⏕ᡂࡍࡿࡢ࡟ᚲせࡔࡗࡓࢱࣥࢼ࣮ࢮࡣࠊ0.3 mg / 10 ml ™

1000 ȣl㸻0.03 mg ࡔࡗࡓࡢ࡛ࠊࡇࢀࢆ 1mgࡢࢱࣥࢼ࣮ࢮࡢሙྜ࡟᥮⟬ࡍࡿ࡜࢞ࣜࢵ

ࢡ㓟⏕ᡂ㔞ࡣ 762.5™10^-4 μmol / 0.03 mg = 2.5 μmol࡛࠶ࡗࡓࠋࢱࣥࢼ࣮ࢮࡢᐃ⩏࡜ࡋ

࡚ࠊpH 5.5ࠊ30Υ࡛ 1ศ㛫࡟ຍỈศゎ࡟ࡼࡗ࡚ 1ȣmol ࡢ࢞ࣜࢵࢡ㓟ࢆ⏕ᡂࡍࡿ㓝⣲㔞

ࢆ 1 unit࡜ࡋࡓሙྜࠊ௒ᅇࡢࢱࣥࢼ࣮ࢮάᛶࡣ 2.5unit ࡛࠶ࡿ࡜ィ⟬ࡉࢀࡓࠋࡋ࠿ࡋ࡞

ࡀࡽࡇࡢ್ࡣࠊ㉎ධ᫬࡟ྠᑒࡉࢀ࡚࠸ࡓ㈨ᩱ࡜ࡣ኱ࡁࡃ␗࡞ࡿ್࡛࠶ࡗࡓࠋ

ࡇࡢࡼ࠺࡟ࠊ௒ᅇࡢࢱࣥࢼ࣮ࢮάᛶ ᐃ࡟࠾࠸࡚ࡣ ᐃ࡟⏝࠸ࡓ⦆⾪ᾮ࡟ၥ㢟Ⅼࡀ࠶

ࡗࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ᐃ࡟⏝࠸ࡓ⦆⾪ᾮࡣ㈨ᩱ࡟ࠕ0.05 M ࢡ࢚ࣥ㓟⦆⾪ᾮࠖ࡜

グ㍕ࡉࢀ࡚࠸ࡓࡀࠊヲ⣽࡞ㄪ〇᪉ἲࡣグ㍕ࡉࢀ࡚࠾ࡽࡎࠊ௚ࡢ㈨ᩱࢆཧ⪃࡟ 0.05 Mࢡ

࢚ࣥ㓟 2Ỉ⣲࣒࢝ࣜ࢘⁐ᾮ࡜ࠊ0.05 M NaOH⁐ᾮࢆΰྜࡋ࡚ㄪ〇ࡋࡓࡀࠊࢼࢺ࣒ࣜ࢘

mol 10

5 . 200 762

5000 100

200 20 mol 50 10

1 .

6 u ⁴ȣ u u u u ⁴ȣ

74

࡞࡝ࡢ㔠ᒓࡣ㓝⣲άᛶ࡟ఱࡽ࠿ࡢᙳ㡪ࢆ୚࠼ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࠊࡑࡢࡓࡵ࡟グ㍕ࡢ

್ࡼࡾࡶపࡃ࡞ࡗࡓࡢ࡛ࡣ࡞࠸࠿࡜⪃࠼ࡽࢀࡓࠋ

ࡋ࠿ࡋ࡞ࡀࡽࠊFig. 4-2 ࡼࡾࠊHPLCศ ᯒ࡛ᚓࡽࢀࡿࢡ࣐ࣟࢺࢢ࣒ࣛ࠿ࡽᕷ㈍ࢱࣥࢼ࣮

ࢮ࡟ࡼࡗ࡚ࢱࣥࢽࣥ㓟୰ࡢ ࢞ ࣟ ࢱ ࣥࢽࣥ 㢮 ࡣ ඘ ศ ࡟ῶ ᑡ ࡋ ࠺ ࡿࡇ ࡜ ࡀ ࢃ ࠿ࡗࡓ ࡢ ࡛ ࠊ ࡇࡢᕷ㈍ࢱࣥࢼ࣮ࢮࢆ⏝࠸࡚࣑ࢬࢼࣛ࠿ࡽᚓࡽࢀࡓᢳฟ≀ࢆฎ⌮ࡍࡿࡇ࡜࡜ࡋࡓࠋ

75

㻜 㻜㻚㻝 㻜㻚㻞 㻜㻚㻟 㻜㻚㻠 㻜㻚㻡 㻜㻚㻢

㻜 㻡 㻝㻜 㻝㻡 㻞㻜 㻞㻡 㻟㻜 㻟㻡

㼙㼕㼚㻚 㻭㼎㼟

Fig. 4-1 Time course of Abs of tannic acid treated by tannase

76

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜

Fig. 5 HPLC chromatograms of tannic acid before treatment by tannase (down side), and after

60 min. treatment by tannase (upper side).

gallotannins gallic acid

gallic acid

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜

Fig. 5 HPLC chromatograms of tannic acid before treatment by tannase (down side), and after

60 min. treatment by tannase (upper side).

gallotannins gallic acid

gallic acid

Fig. 4-2 HPLC chromatograms of tannic acid

before (a) and after (b) 60min treated by tannase

77

㻲㼕㼓㻚㻌㻠㻙㻟㻌㻯㼔㼍㼚㼓㼑㻌㼛㼒㻌㼓㼍㼘㼘㼕㼏㻌㼍㼏㼕㼐㻌㼍㼙㼛㼡㼚㼠㻌㼜㼞㼛㼐㼡㼏㼑㼐㻌㼒㼞㼛㼙 㻌㼠㼍㼚㼚㼕㼏㻌㼍㼏㼕㼐㻌㼔㼥㼐㼞㼛㼘㼥㼟㼕㼟㻌㼠㼞㼑㼍㼠㼙㼑㼚㼠㻌㼎㼥㻌㼠㼍㼚㼚㼍㼟㼑 㼥㻌㻩㻌㻜㻚㻝㻞㻝㻠㼤㻌㻙㻌㻜㻚㻢㻝㻡㻥

㻾

^㻞

㻌㻩㻌㻜㻚㻥㻤㻣㻝

㻜㻚㻜 㻝㻚㻜 㻞㻚㻜 㻟㻚㻜 㻠㻚㻜 㻡㻚㻜 㻢㻚㻜 㻣㻚㻜

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

㼠㼕㼙㼑㻌㻔㼙㼕㼚㻚㻕

㼍㼙 㼛 㼡 㼚 㼠㻌㼛 㼒㻌㼓 㼍㼘 㼘㼕㼏 㻌㼍 㼏 㼕㼐㻌㻔 μ 㼓㻕

78

㻲㼕㼓㻚㻌㻠㻙㻠㻌㻭㼚㼍㼘㼥㼠㼕㼏㼍㼘㻌㼏㼡㼞㼢㼑㻌㼛㼒㻌㼓㼍㼘㼘㼕㼏㻌㼍㼏㼕㼐㻌㼍㼚㼐㻌㼜㼑㼍㼗㻌㼍㼞㼑㼍㻌㼛㼚 㻴㻼㻸㻯㻌㼏㼔㼞㼛㼙㼍㼠㼛㼓㼞㼍㼙

㼥㻌㻩㻌㻟㻱㻗㻜㻢㼤 㻾^㻞㻌㻩㻌㻜㻚㻥㻥㻝㻟

㻜㻚㻜㻱㻗㻜㻜 㻞㻚㻜㻱㻗㻜㻢 㻠㻚㻜㻱㻗㻜㻢 㻢㻚㻜㻱㻗㻜㻢 㻤㻚㻜㻱㻗㻜㻢 㻝㻚㻜㻱㻗㻜㻣 㻝㻚㻞㻱㻗㻜㻣 㻝㻚㻠㻱㻗㻜㻣 㻝㻚㻢㻱㻗㻜㻣

㻜 㻝 㻞 㻟 㻠 㻡 㻢

㼏㼛㼚㼏㼑㼚㼠㼞㼍㼠㼕㼛㼚㻌㼛㼒㻌㼓㼍㼘㼘㼕㼏㻌㼍㼏㼕㼐㻔μ 㼓㻕 㼍㼞㼑㼍

79

4-3-2 ೺඲࣑ࢬࢼࣛ㎶ᮦᢳฟ≀ 㓑㓟࢚ࢳࣝྍ⁐㒊㸦NS-EtOAc-S㸧ࡢ ㄪ〇

580.0gࡢ೺඲࣑ࢬࢼࣛ㎶ᮦ࠿ࡽᚓࡽࢀࡓ NS-eࡢᾮᾮศ㓄࡟ࡼࡗ࡚ࠊ࢚࣮ࢸࣝྍ⁐㒊

0.9gࠊ㓑㓟࢚ࢳࣝྍ⁐㒊 2.59gࠊn-ࣈࢱࣀ࣮ࣝྍ⁐㒊 3.17gࠊn-ࣈࢱࣀ࣮ࣝ୙⁐㒊 2.64g

ࢆᚓࡓࠋࡑࢀࡽ࡟ࡘ࠸࡚ HPLCศᯒࢆ⾜ࡗࡓ࡜ࡇࢁ(Fig. 4-5)ࠊNS-e࡛ぢࡽࢀࡿࠊ➨୍

❶ࡢ 1-3-4࡛♧ࡋࡓࡼ࠺࡟㓟ຍỈศゎ࡛㢧ⴭ࡟ῶᑡࡋࡓࠊຍỈศゎᛶࢱࣥࢽࣥ࡜ᛮࢃ

ࢀࡿࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 30 ศ௜㏆ࡢࣈ࣮ࣟࢻ࡞ࣆ࣮ࢡ⩌ࡀࠊ࢚࣮ࢸࣝྍ⁐㒊࡛ࡣ࠿

࡞ࡾศ㞳⬟ࡼࡃศ࠿ࢀ᳨࡚ฟࡉࢀࡓࠋࡋ࠿ࡋࠊࡇࡢ࠺ࡕࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 31ศ௜

㏆࡟ぢࡽࢀࡿࣆ࣮ࢡࡣࡑࡢࣜࢸࣥࢩࣙࣥࢱ ࢖࣒࡜ UVࢫ࣌ࢡࢺࣝࡀ࢚ࣛࢢ㓟ࡢࡶࡢ࡜

୍⮴ࡋࡓࠋࡇࡢࡇ࡜࠿ࡽࠊ࢚࣮ࢸࣝྍ⁐㒊࡟ࡣ୺࡟ࠊ࢚ࣛࢢ㓟ࡀྵࡲࢀࡿ࡜⪃࠼ࡽࢀ

ࡓࠋ୍᪉ࠊࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 30 ศ௜㏆ࡢࣆ࣮ࢡ⩌ࡣࠊ㓑㓟࢚ࢳࣝྍ⁐㒊ࠊn-ࣈࢱࣀ

࣮ࣝྍ⁐㒊ࠊn-ࣈࢱࣀ࣮ࣝ୙⁐㒊࡟ศ㓄ࡉࢀ࡚࠸ࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊࣆ࣮ࢡࡢᙧ≧࠿

ࡽࠊ᭱ࡶຍỈศゎᛶࢱࣥࢽࣥ㢮ࢆ୺࡟ྵࡴࡢࡣ㓑㓟࢚ࢳࣝྍ⁐㒊࡛࠶ࡗࡓࠋࡇࡢࡇ࡜

࠿ࡽࠊẚ㍑ⓗ㔞ࡶከࡃࠊຍỈศゎᛶࢱࣥࢽࣥࢆከࡃྵࡴ㓑㓟࢚ࢳࣝྍ⁐㒊

㸦NS-EtOAc-S㸧ࢆ೺඲࣑ࢬࢼࣛ㎶ᮦ⏤᮶ࡢຍỈศゎᛶࢱࣥࢽࣥ㢮࡜ࡋ࡚ࠊ௒ᚋࡢ᳨ウ

࡛౑⏝ࡍࡿࡇ࡜࡜ࡋࡓࠋ

80

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

Fig. 4-5 HPLC chromatograms of 70% aqueous acetone extracts (a), ether soluble part (b), ethyl acetate soluble part (c), n-BuOH soluble part (d) and n-BuOH insoluble part (e)

㼏㻕

㼎㻕

㼍㻕 㼑㻕

㼐㻕

81

4-3-3 NS-EtOAc-Sࡢᕷ㈍ࢱࣥࢼ࣮ࢮฎ⌮࡟ࡼࡿ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛࡢኚ໬

R. quercivora ࡀឤᰁࡋࡓ࣑ࢬࢼ࡛ࣛࡣࠊࡑࡢ㎶ᮦࡢኚⰍ㒊఩࡟࠾࠸࡚೺඲࡞࣑ࢬࢼࣛ

㎶ᮦ࡟ྵࡲࢀࡿຍỈศゎᛶࢱࣥࢽࣥ㢮ࡢຍỈศゎ≀࡛࠶ࡿࠊ࢞ࣜࢵࢡ㓟ࡸ࢚ࣛࢢ㓟ࡀ

᳨ฟࡉࢀࡓࠋࡲࡓࠊຍỈศゎᛶࢱࣥࢽࣥ㢮ࡢຍỈศゎ㓝⣲࡜ࡋ࡚ࡣࠊࢱࣥࢼ࣮ࢮࡀ▱

ࡽࢀ࡚࠸ࡿࠋࡑࡇ࡛ࠊᕷ㈍ࡢࢱࣥࢼ࣮ࢮࢆ㸦A. oryzae ⏤᮶㸧⏝࠸࡚ᐇ㝿ࡢᢳฟ≀ࢆ㓝

⣲ฎ⌮ࡋࠊHPLCࢡ࣐ࣟࢺࢢ࣒ࣛศᯒࢆ⾜ࡗ࡚ࠊᐇ㝿࡟⿕ᐖᮌࡢኚⰍࡋࡓ㎶ᮦ࡛ ぢࡽ

ࢀࡿࡼ࠺࡟ࠊ࢞ࣜࢵࢡ㓟࠾ࡼࡧ࢚ࣛࢢ㓟ࡢ⏕ᡂࡀ㉳ࡇࡿ࠿࡝࠺࠿ࢆ☜ㄆࡍࡿᐇ㦂ࢆ⾜

ࡗࡓࠋNS-EtOAc-Sࢆࢱࣥࢼ࣮ࢮ࡟ࡼࡗ࡚ฎ⌮ࡋࠊฎ⌮๓ᚋ࡟࠾ࡅࡿ HPLCࢡ࣐ࣟࢺ

ࢢ࣒ࣛࢆẚ㍑ࡋࡓࠋࡑࡢ⤖ ᯝ ࠊ ຍ Ỉศゎ ᛶ ࢱ ࣥ ࢽ ࣥ㢮 ࡟ ┦ ᙜ ࡍࡿ ࣆ ࣮ ࢡ 㢮ࡢῶ ᑡ ࡀ ࠊ

࠶ࡲࡾぢࡽࢀ࡞࠿ࡗࡓ(Fig. 4-6)ࠋ࢞ࣜࢵࢡ㓟ࡣ㢧ⴭ࡞ቑຍࡀぢࡽࢀࡓࡀࠊ࢚ࣛࢢ㓟࡟

┦ᙜࡍࡿࣆ࣮ࢡࡣ᳨ฟࡉࢀ࡞࠿ࡗࡓࠋࡲࡓࠊ➨୍❶ࡢ 1-3-4࡛♧ࡋࡓ㏻ࡾࠊNS-eࡣ㓟

ຍỈศゎ࡟ࡼࡗ࡚࢚ࣛࢢ㓟ࢆ⏕ࡌࡿࡇ࡜࠿ࡽࠊNS-e࡜ࡑࡢຍỈศゎᛶࢱࣥࢽࣥ㢮ࢆ୺

࡟ྵࢇ࡛࠸ࡿ࡜ᛮࢃ ࢀࡿ NS-EtOAc-S ࡟ࡣ࢚ࣛࢪࢱࣥࢽࣥ㢮ࡀྵࡲࢀࡿ࡜⪃࠼ࡽࢀࡿࠋ

௨ୖࡢ஦࠿ࡽࠊࡇࡢ⦆⾪ᾮ୰ࡢฎ⌮࡟࠾࠸࡚ࢱࣥࢼ࣮ࢮࡣ࢞ࣟࢱࣥࢽࣥࢆศゎࡍࡿࡶ

ࡢࡢࠊ࢚ࣛࢢࢱࣥࢽࣥࡢศゎ⬟ࢆ♧ࡉ࡞࠸࡜ゝ࠼ࡓࠋ ࡋ࠿ࡋ࡞ࡀࡽࠊ➨୍❶ 1-3-3࡛

♧ࡋࡓ㏻ࡾࠊ⿕ᐖᮌࡢኚⰍࡋࡓ㎶ᮦ࡟ࡣ࢚ࣛࢢ㓟ࡀከࡃྵࡲࢀ࡚࠸ࡓࡇ࡜࠿ࡽࠊ࡞ࢇ

ࡽ࠿ࡢ⌮⏤࡛NS-e࡟ྵࡲࢀࡿ࢚ࣛࢪࢱࣥࢽࣥ㢮ࡣຍỈศゎࡍࡿ࡜ᛮࢃࢀࡓࠋࡑࡇ࡛ࠊ

ឤᰁࡋࡓ R. quercivoraࡀ⏕ᡂࡍࡿ㓝⣲࡟ࡣ࢚ࣛ ࢪࢱࣥࢽࣥศゎ⬟ࡀ࠶ࡿࡢ࡛ࡣ࡞࠸࠿

࡜⪃࠼ࠊPGG ศゎࡢ⤒᫬ኚ໬ࢆ㏣ࡗࡓᐇ㦂࡜ ྠࡌࡼ࠺࡟ᾮయᇵᆅ࡟ EtOAc-Sࢆῧຍࡋࠊ

ࡇࡢᇵᆅ࡛ R. quercivoraࢆᇵ㣴ࡋ࡚ࠊᇵᆅᡂศࡢ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛࢆẚ㍑ࡍࡿࡇ

82

࡜࡟ࡋࡓࠋ

83

㼓㼍㼘㼘㼕㼏 㼍㼏㼕㼐

Retention time (min)

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

㼙㼕㼚㻚

Fig. 4-6 HPLC chromatograms of EtOAc-S (a) treated with commercial tannase (b) blank.

a)

b)

84

4-3-4 NS-EtOAc-Sࡢࢼࣛ⳦⢒㓝⣲ฎ⌮࡟ࡼࡿ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛࡢኚ໬

ᕷ㈍ࢱࣥࢼ࣮ࢮࡀ NS-EtOAc-S࡟ྵࡲ ࢀࡿ࢚ࣛࢪࢱࣥࢽࣥ㢮ࢆຍỈศゎࡋ࡞࠿ࡗࡓࡇ

࡜࠿ࡽࠊࢼࣛ⳦⢒㓝⣲࡟ࡼࡿຍỈศゎࢆヨࡳࡓࠋNS-EtOAc-Sࢆῧຍࡋࡓᇵᆅ࡛ࢼࣛ

⳦ࢆᇵ㣴ࡋࠊᇵ㣴ᚋ࡟ᇵᆅࡢ୍㒊ࢆᅇ཰ࡋࠊHPLCศᯒࡋࡓ(Fig. 4-7)ࠋࡑࡢ ⤖ᯝࠊࢼ

ࣛ⳦ࡢ᥋✀࡟ࡼࡗ࡚ࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 30ศ௜㏆࡟ぢࡽࢀࡿຍỈศゎᛶࢱࣥࢽࣥ࡜

ᛮࢃࢀࡿࣆ࣮ࢡࡀ㢧ⴭ࡟ῶᑡࡋࠊࡲࡓࠊR.T. 32 min.௜㏆࡟ẚ㍑ⓗࢩ࣮ࣕࣉ࡞ࣆ࣮ࢡࡀ

᳨ฟࡉࢀࡓࠋࡇࡢࣆ࣮ࢡࡣ࢚ࣛࢢ㓟࡜ྠࡌࣜࢸࣥࢩࣙࣥࢱ࢖࣒ ࡜ UVࢫ࣌ࢡࢺࣝࢆ♧

ࡋࡓࡇ࡜࠿ࡽࠊ࢚ࣛࢢ㓟ࡀ᳨ฟࡉࢀࡓ࡜⪃࠼ࡽࢀࡓࠋ௨ୖࡢࡇ࡜࠿ࡽࠊR. quercivora 㸦ࡢ⢒㓝⣲㸧࡟ࡣ࢚ࣛࢢࢱࣥࢽࣥࡢศゎ⬟ࡀ࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ㏻ᖖࡢࢱࣥࢼ࣮ࢮࡣ

ຍỈศゎᛶࢱࣥࢽࣥࡢ୰࡛ࡶ࢞ࣟࢱࣥࢽࣥࢆศゎࡍࡿ㓝⣲࡛࠶ࡿࡢ࡛ࠊR. quercivora ࡀ⏕ᡂࡍࡿࢱࣥࢼ࣮ࢮࡣ≉ᚩⓗ ࡞ᛶ㉁ࢆ᭷ࡋ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࠋ

ࡲࡓࠊࣜࢸࣥࢩࣙࣥࢱ࢖࣒ 10 ศ࡟ぢࡽࢀࡿࡣࡎࡢ࢞ࣜࢵࢡ㓟ࡢࣆ࣮ࢡࡣࠊ࡯ࡰ᳨ฟ ࡉࢀ࡞࠿ࡗࡓࠋࡇࢀࡣ➨஧❶࡛㏙࡭ࡓࡼ࠺࡟ࠊ࢞ࣜࢵࢡ㓟ࡣࢼࣛ⳦ࡢᇵ㣴ᙜึࡣࡑࡢ

㔞ࢆቑຍࡉࡏࡿࡶࡢࡢࠊࡑࡢᇵ㣴࡟కࡗ࡚ࢼࣛ⳦ࡀࣛࢵ࣮࢝ࢮࢆᨺฟࡍࡿࡓࡵ࡟㔜ྜ

ࡋࡓࡓࡵ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ

85

㻜 㻝㻜 㻞㻜 㻟㻜 㻠㻜 㻡㻜 㻢㻜

㼙㼕㼚㻚

a)

b)

ellagic acid

Fig. 4-7 HPLC chromatograms of EtOAc-S (a) treated with crude enzyme of R. quercivora. (b) blank.

Retention time (min)

86

4-3-5 NS-EtOAc-Sῧຍᇵᆅ࡟࠾ࡅࡿࠊࢼࣛ⳦ࡢ⳦య⾲㠃ࡢኚ໬

NS-EtOAc-Sῧຍᇵᆅ࡛ࢼࣛ⳦ࢆᇵ㣴ࡋࡓ࡜ࡇࢁࠊᇵ㣴๓ࡣⓑⰍࡔࡗࡓ⳦యࡀࠊ⫗║

࡛ぢ࡚ࡶ〓Ⰽ࡟࡞ࡗ࡚࠸ࡿࡇ࡜ࡀࢃ࠿ࡗࡓࠋࡑࡇ࡛ࠊ⳦యࡢ ୍㒊ࢆගᏛ㢧ᚤ㙾࡛ほᐹ

ࡋࡓ(Photo)ࠋࡑࡢ⤖ᯝࠊᇵ㣴๓࡟ࡣࢼࣛ⳦ࡢ⳦యࡣ㏱࡛᫂ࠊ⳦య⾲㠃ࡶพฝࡀ࡞ࡃ⁥

ࡽ࠿࡟ぢ࠼ࡿࡀࠊᇵ㣴ᚋ࡟ࡣࡑࡢ⾲㠃࡟〓Ⰽࡢ⢏Ꮚ≧ࡢ≀㉁ࡀከᩘ௜╔ࡋ࡚࠸ࡓࠋࡇ

ࢀࡀ NS-EtOAc-S࡛࠶ࡿ࠿࡝࠺࠿ࡣࠊ⌧᫬Ⅼ࡛ࡣࢃ࠿ࡽ࡞࠸ࠋࡋ࠿ࡋࠊ➨஧❶ࡢ 2-3

࡛㏙࡭ࡓࡼ࠺࡟ࠊࣔࢹࣝ໬ྜ≀࡜ࡋ࡚⏝࠸ࡓࢱࣥࢽࣥ㓟ࡢሙྜࠊ⳦యࢆྵࢇࡔࢼࣛ⳦

ᇵ㣴ᇵᆅ࡟ῧຍࡋࡓሙྜࡢ ᪉ ࡀ ࢱ ࣥࢽࣥ 㓟 ࡟ ヱ ᙜ ࡍࡿ ࣆ ࣮ ࢡ ࡣⴭ ࡋ ࡃ ῶ ᑡࡋ࡚ ࠸ ࡓ ࠋ ௨ୖࡢࡇ࡜࠿ࡽࠊࢼࣛ⳦ࡣ⳦యእ࡟ࢱࣥࢼ࣮ࢮᵝࡢ㓝⣲ࢆᨺฟࡍࡿࡀࠊࡑࡢ୍㒊ࡣ⳦

య⾲㠃࡟࡜࡝ࡲࡾࠊࡑࡇ࡛㔜Ⅼⓗ࡟ຍỈศゎᛶࢱࣥࢽࣥ㢮ࡢศゎࡀ㉳ࡇࡗ࡚࠸ࡿྍ⬟

ᛶࡀ࠶ࡿࠋ

87

Photo Fungus body of R. quercivora before (a, ™100) and after (b, ™200) incubation with NS-EtOAc-S

a)

b)

88

4-4 ࡲ࡜ࡵ

೺඲࡞࣑ࢬࢼࣛࡢᢳฟ≀ࢆྛ✀᭷ᶵ⁐፹࡟࡚ᾮᾮศ㓄ࡋࡓ࡜ࡇࢁࠊ୺࡟ྵࡲࢀࡿຍỈ ศゎᛶࢱࣥࢽࣥ㢮ࡣ㓑㓟࢚ࢳࣝ⏬ศ࡟ศ㓄ࡉࢀࡓࠋࡇࡢ㓑㓟࢚ࢳࣝྍ⁐㒊ࢆᕷ㈍ࢱࣥ

ࢼ࣮ࢮ࡛ฎ⌮ࡋࡓ࡜ࡇࢁࠊᑡ㔞ࡢ࢞ࣜࢵࢡ㓟ࢆ᳨ฟࡋࡓࡀࠊຍỈศゎᛶ ࢱࣥࢽࣥ㢮ࢆ

♧ࡍࣆ࣮ࢡࡣ࠶ࡲࡾῶᑡࡋ࡞࠿ࡗࡓࠋ୍᪉ࠊࢼࣛ⳦ࡢ⢒㓝⣲࡛ฎ⌮ࡋࡓሙྜࠊຍỈศ

ゎᛶࢱࣥࢽࣥ㢮ࡣ㢧ⴭ࡟ῶᑡࡋࠊ࢚ࣛࢢ㓟ࡢࣆ࣮ࢡࡀ᳨ฟࡉࢀࡓࠋ ࡲࡓࠊࡇࡢຍỈศ ゎ཯ᛂࡣࠊࢼࣛ⳦ࡢ⳦య⾲㠃㏆ࡃࢆ୰ᚰ࡟㉳ࡇࡗ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࠋ

ୖグࡢෆᐜࡣ 2008ᖺ࡟ Journal of wood science ࡟࡚Ⓨ⾲ࡋࡓ ²⁾ࠋ

89

4-5 ཧ⪃ᩥ⊩

1. ㉥ᯇ ໏㸪ୖᔱ ὒ㸪⸨஭ ฼㑻㸪ྜྷཎ ୍ᖺ. 1985. ࣜࢢࢽࣥศゎ⬟ࡢᙉຊ࡞ⓑ

Ⰽ⭉ᮙ⳦ࡢࢫࢡ࣮ࣜࢽࣥࢢ. ⣬ࣃᢏㄅ 39:65-74

2. Shashi Sharma, Lata Agarwal, Rajendra Kumar axena. 2008. Purification, immobilization and characterization of tannase from Penicillium cariable. Bioresource Technology.

99:2544-2551

2. K. Imai , T. Mitsunaga , H. Takemoto , T. Yamada, S. Ito, H. Ohashi (2008) Extractives of Quercus crispula sapwood infected by the pathogenic fungi Raffaelea

quercivora (I) : Comparison of sapwood extractives non-infected and infected. J.Wood. Sci.

55:126-132

90

➨ 5❶ ࢼࣛᯤࢀ⿕ᐖࢆཷࡅࡓ࣑ࢬࢼࣛ㎶ᮦ࠿ࡽࡢᡂศࡢ༢㞳ྠᐃ

5-1 ⥴ゝ

࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධ࡟ࡼࡗ࡚ࠊ࣑ࢬࢼࣛࡢ㎶ᮦ࡛ࡣࠊຍỈศゎᛶࢱࣥࢽࣥ㢮 ࡀࢼࣛ⳦ࡢㄏᑟࡍࡿ㓝⣲ᵝ≀㉁࡟ࡼࡗ࡚ຍỈศゎࢆཷࡅࠊࡑࡢ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿ࢞ࣜ

ࢵࢡ㓟࡜࢚ࣛࢢ㓟ࡀ⏕ᡂࡍࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊ➨୍❶ࡢ 1-3-3 ࡛♧ࡋ ࡓ HPLCࢡ࣐ࣟࢺࢢ࣒ࣛ࠿ࡽࠊ௒ᅇධᡭࡋࡓ࣑ࢬࢼࣛࡢ⿕ᐖᮦ࡛ࡣࠊຍỈศゎᛶࢱࣥ

ࢽࣥ㢮ࡀࢃࡎ࠿࡟ṧᏑࡋ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡗࡓࠋࡇࢀࡽࡢຍỈศゎᛶࢱࣥࢽࣥ㢮ࡣ ຍ Ỉศゎࢆཷࡅ࡚࠸ࡿ㏵୰ࡢ≧ែ࡛࠶ࡿ࠿ ࠊ࠶ࡿ࠸ࡣࢼࣛ⳦ࡢᨺฟࡍࡿ㓝⣲ᵝ≀㉁࡟ࡼ

ࡗ࡚㞴ศゎᛶ࡛࠶ࡿྍ⬟ᛶࡀ࠶ࡿࠋ

ࡲࡓࠊࡇࢀࡲ࡛࡟ Quercusᒓࡢᢳฟᡂศ࡟ࡘ࠸࡚ࡣࠊQ. cocciferaࡸ Q. suber࡞࡝࡛ຍ

Ỉศゎᛶࢱࣥࢽࣥ㢮ࡢ᥈⣴ࡀࡉࢀ࡚࠸ࡿࡀ ¹⁾ࠊ࣑ࢬࢼࣛࡢ㎶ᮦ࡟ᑐࡍࡿᢳฟᡂศࡢ༢ 㞳ྠᐃࡣ⾜ࢃࢀ࡚࠸࡞࠸ࠋ

ࡑࡇ࡛ࠊ࢝ࢩࣀࢼ࢞࢟ࢡ࢖࣒ࢩࡢ✸ධࢆཷࡅᯤṚࡋࡓ࣑ࢬࢼࣛࡢ㎶ᮦᢳฟ≀࡟ྵ᭷ࡋ

࡚࠸ࡿᡂศࡢ᥈⣴ࢆ⾜࠺ࡇ࡜࡜ࡋࡓࠋ

91

ドキュメント内 ナラ枯れ原因菌 Raffaelea quercivora 侵入に応答するミズナラの抽出成分に関する研究 (ページ 69-94)