㠽ข⋵↥ᬺᛛⴚ࠮ࡦ࠲࡯‫⎇ޓ‬ⓥႎ๔ͳ  ਇቢో⩶Trichoderma harzianumߩၭ㙃ࠈᶧࠍ↪޿ߚ㕖᥏ൻࠠ࠴ࡦ߆ࠄߩN-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩല₸⊛↢↥Effective Production of N-Acetyl-D-glucosamine from Decrystalized Chitin by Culture Filtrates of Trichoderma harzianumศ↰᤯৻Shinichi Yoshidaࡅࠕ࡞ࡠࡦ㉄วᚑଦㅴ⢻߿᛫Ἳ∝૞↪ߥߤ߇ႎ๔ߐࠇ‫ޔ‬ᯏ⢻ᕈ↞๧ᢱߣߒߡᵈ⋡ࠍ㓸߼ߡ޿ࠆ N-ࠕ࠮࠴࡞-D-ࠣ
࡞ࠦࠨࡒࡦߩᄢ㊂↢↥ᣇᴺߩ㐿⊒ࠍ⋡⊛ߣߒߡ‫ޔ‬ਇቢో⩶ Trichoderma harzianum ߩ↥↢ߔࠆࠠ࠴ࡦಽ⸃㉂⚛ࠍ↪
޿ߡࠞ࠾Ზ↱᧪ߩࠠ࠴ࡦࠍಽ⸃ߔࠆᣇᴺߦߟ޿ߡᬌ⸛ࠍട߃ߚ‫ޔߕ߹ޕ‬⒳‫ߩޘ‬὇⚛Ḯ‫⚛⓸ޔ‬Ḯࠍ↪޿ߡ T. harzianumߦࠃࠅ↥↢ߐࠇࠆࠠ࠴ࡦಽ⸃㉂⚛ᵴᕈߩ⚻ᤨᄌൻߦߟ޿ߡᬌ⸛ࠍട߃ߚߣߎࠈ‫ޔ‬὇⚛Ḯߣߒߡ☳ᧃ⁁ࠠ࠴ࡦ‫⚛⓸ޔ‬Ḯߣߒߡ⎫㉄ࠕࡦࡕ࠾࠙ࡓࠍ↪޿ߚ႐วߦࠠ࠴࠽࡯࠯ߣ N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ߩ㜞޿ᵴᕈ߇ⷰኤߐࠇߚ‫ޕ‬ߒ߆ߒߥ߇ࠄ‫ޔ‬ၭ㙃ᶧਛߦ N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩሽ࿷ߪ⹺߼ࠄࠇߕ‫ߪߣߎߩߎޔ‬㉂⚛ಽ⸃ߦࠃࠅ↢ᚑߒߚ
N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߪ T. harzianum ߦࠃࠅઍ⻢ߐࠇߚߚ߼ߣ⠨߃ࠄࠇߚ‫⩶ޔߢߎߘޕ‬૕ࠍ㒰෰ߒߚၭ㙃ࠈᶧࠍ࿁෼ߒࠠ࠴ࡦߣߩ෻ᔕࠍ⹜ߺ‫ޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩ↢ᚑ㊂ߦߟ޿ߡᬌ⸛ࠍട߃ߚߣߎࠈ‫ߩ⁁ᧃ☳ޔ‬ࠠ࠴ࡦߢߪߘߩ↢ᚑߪ߶ߣࠎߤⷰኤߐࠇߥ߆ߞߚ߇‫ޔ‬޽ࠄ߆ߓ߼ࠕ࡞ࠞ࡝ಣℂߦࠃࠅ⚿᥏᭴ㅧࠍ⎕უߒߡ߅޿ߚ㕖᥏ൻࠠ࠴ࡦߪቢోߦಽ⸃ߐࠇ‫ޔ‬48 ᤨ㑆એౝߢ߶߷ 100%ߩ෼₸ߢ⋡⊛‛߳ߣᄌ឵ߐࠇࠆߎߣ߇ಽ߆ߞߚ‫ޕ‬

N-Acetyl-D-glucosamine is one of amino sugars, and is recently reported to stimulate the production of hyaluronan in animaltissues and to possess a unique range of anti-inflammatory activities. In order to develop large-scale production method ofN-acetyl-D-glucosamine applicable to food industries, hydrolysis of chitin from crab shells by chitinolytic enzymes ofdeuteromycete Trichoderma harzianum was investigated. Incubation of T. harzianum with chitin as carbon source andammonium sulfate as nitrogen source leads to the production of high activity of chitinolytic enzymes, chitinase andN-acetylhexosaminidase.

However, no N-acetyl-D-glucosamine was detected in the culture medium of T. harzianum,indicating that the N-acetyl-D-glucosamine produced in the medium was metabolized by T. harzianum. Therefore, the culture
filtrates containing chitinolytic enzymes were collected to remove mycelia of T. harzianum, and the incubation with powderedchitin or decrystallized chitin was attempted. Powdered chitin was not hydrolyzed by the culture filtrates of T. harzianum, but,decrystallized chitin was completely hydrolyzed to give ca. 100% yield of N-acetyl-D-glucosamine within 48 hours.

 ߪߓ߼ߦ૞↪ߦ㑐ߔࠆ⎇ⓥ߇ᐢ▸ߦⴕࠊࠇ‫㉄ࡦࡠ࡞ࠕࡅޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߪ‫࠾ࠞޔ‬Ზ߿ࠛࡆߩวᚑଦㅴലᨐ 2)߿᛫Ἳ∝૞↪ 3)ߥߤ߇ႎ๔ߐࠇ‫ޔ‬Ზ‫ޔ‬᣸⯻㘃ߦ฽߹ࠇࠆࠠ࠴ࡦ߿േ‛⚦⢩ਛߦ฽߹ஜᐽ㘩ຠ‫ޔ‬ൻ♆ຠේᢱ‫ޔ‬ᯏ⢻ᕈ↞๧ᢱߣߒߡᵈ⋡ࠇࠆࡅࠕ࡞ࡠࡦ㉄߿ࠦࡦ࠼ࡠࠗ࠴ࡦ⎫㉄ߥߤߩࠍ㓸߼ߡ޿ࠆ‫ޕ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߪ‫ޔ‬ࠣ࡞ࠦࠨࡒࡁࠣ࡝ࠞࡦࠍ᭴ᚑߒߡ޿ࠆࠕࡒࡁၮ

D-ࠣ࡞ࠦࠨࡒࡦࠍ N-ࠕ࠮࠴࡞ൻߔࠆߎߣߦࠃࠅ

ࠍ᦭ߔࠆන♧㘃ߩ৻⒳ߢ޽ࠅ‫⥄ޔ‬ὼ⇇ߦᐢߊಽᏓൻቇ⊛ߥᣇᴺߢኈᤃߦวᚑߔࠆߎߣ߇ߢ߈ࠆ߇ 4)‫ޔ‬ߔࠆ 1)‫ޕ‬ㄭᐕ‫ޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩ↢ℂஜᐽ㘩ຠ߿ᯏ⢻ᕈ↞๧ᢱߣߒߡ೑↪ߔࠆߦߪ㘩̶1̶ຠⴡ↢ᴺߦḰ᜚ߒߚ‫ޔ‬ᄙ♧㘃ߢ޽ࠆࠠ࠴ࡦߩട᳓ᵞᵺᶧ߇ਛᕈߦߥࠆ߹ߢᔀᐩ⊛ߦᵞᵺߒߚ‫⵾⺞ޕ‬ಽ⸃ࠍⴕ޿ಽ⸃‛߆ࠄಽ㔌࡮♖⵾ߔࠆᣇᴺࠍᬌ⸛ߒߚࠥ࡞⁁ߩ㕖᥏⾰ࠠ࠴ࡦߩ৻ㇱࠍߪ߆ࠅขࠅ‫ޔ‬ߔࠆᔅⷐ߇޽ࠆ 5)‫ޕ‬110͠ߩᕡེ᷷ਛߢੇ῎ࠍⴕ޿ߘߩ฽᳓₸ࠍ᷹ቯࠞ࠾Ზ߆ࠄᓧࠄࠇࠆࠠ࠴ࡦߩട᳓ಽ⸃ߦߪ㉄‫ޕ‬⺞⵾ߒߚ⹜ᢱߩ㨄✢࿁᛬ߪ‫ޔ‬ߒߚ㧔฽᳓₸ 92.6㧑㧕ࠍ↪޿ࠆᣇᴺߣട᳓ಽ⸃㉂⚛ࠍ↪޿ࠆᣇᴺ߇᜼ℂቇ㔚ᯏ㧔ᩣ㧕⵾㨄✢࿁᛬ⵝ⟎ RINT 2500 ࠍ↪޿ߍࠄࠇࠆ‫ࠆ޿↪ࠍ㉄ޕ‬ᣇᴺߢߪ‫࡞࠴࠮ࠕޔ‬ၮ߽หߡ᷹ቯߒߚ‫ ↪૶ޕ‬X ✢ߪ CuKǩ✢ࠍ↪޿‫▤ޔ‬㔚࿶ᤨߦ⣕㔌ߒ D-ࠣ࡞ࠦࠨࡒࡦ߇ᄢ㊂ߦ↢ᚑߔࠆߚ߅ࠃ߮▤㔚ᵹߪ 50 kV‫ޔ‬300 mA ߣߒߚ‫ޔ߅ߥޕ‬ታ߼‫࡞࠴࠮ࠕޔ‬ၮ߇⣕㔌ߒߥ޿෻ᔕ᷷ᐲ߿ᤨ㑆ߩ෩㛎ߦ߽ߜ޿ߚ☳ᧃࠠ࠴ࡦߩ฽᳓₸ߪ㧢㧑ߢ޽ࠆ߇‫ޔ‬߹ߚ‫ޔ‬ኒߥ⺞ᢛ߿ಽ㔌࡮♖⵾ᠲ૞߇ᔅⷐߢ޽ࠆ 6)‫ޕ‬X ✢᷹ቯߩ㓙‫ޔ‬㕖᥏ൻࠠ࠴ࡦߩ᷹ቯ᧦ઙߦㄭߠߌ

Ꮏ႐⃻႐ߢߪ㉄ߦࠃࠆ⸳஻ߩഠൻ߿㉄ᑄᶧಣℂ

ࠆߚ߼ X ✢↪⹜ᢱߪ᳓ࠍ฽߹ߖߡ 75.7%ߣߒߚ‫ޕ‬ߦⷐߔࠆࠦࠬ࠻߇໧㗴ߣߥࠆ‫৻ޕ‬ᣇ‫ࡦ࠴ࠠޔ‬ട᳓ಽ⸃㉂⚛ࠍ↪޿ߚᣇᴺߩᬌ⸛߽ⴕࠊࠇߡ߅ࠅ‫ࠠޔ‬ 6TKEJQFGTOCJCT\KCPWO ߩၭ㙃࠴ࡦಽ⸃ᓸ↢‛ߩᬌ⚝߿ಽ⸃㉂⚛ߩ↢↥᧦ઙߦ ೨ၭ㙃ߟ޿ߡ⎇ⓥߐࠇߡ޿ࠆ‫ޕ‬ේᢱߣߥࠆࠠ࠴ࡦߩ㜞ᐲ

D-ࠣ࡞ࠦ࡯ࠬ㧔㧞㧑㧕‫ࡦ࠻ࡊࡍ࡝ࡐޔ‬㧔0.5㧑㧕‫ޔ‬ߥ⚿᥏ᕈߩ໧㗴߆ࠄࠗࠞ↱᧪ߩǪ-ࠠ࠴ࡦࠍ಴⊒‫ޔ‬MgSO㧠࡮㉂Უࠛࠠࠬ㧔0.2㧑㧕
‫ޔ‬KH2PO4㧔0.㧝㧑㧕

ේᢱߣߔࠆᣇᴺ߇ឭ᩺ߐࠇߡ޿ࠆ߇ 7㧕‫ޔ‬Ꮏᬺ⊛ߦ

7H2O㧔0.05 㧑㧕߅ࠃ߮ኙᄤ㧔㧞㧑㧕ࠍ฽߻ᐔ㕙ណ▚ߩว߁㊂ߩේᢱ⏕଻ࠍ⠨߃ࠆߣ‫ࡆࠛ߿࠾ࠞޔ‬ၭ࿾ߦ‫⩶ޔ‬૕ࠍ㧝⊕㊄⡊ಽធ⒳ߒ‫ޔ‬30͠ߢ㧠ᣣ㑆↱᧪ߩǩ-ࠠ࠴ࡦࠍ૶߁ߩ߇ℂᗐߢ޽ࠆ߇‫⚳ᦨޔ‬ၭ㙃ߒߚ‫ޕ‬෼₸ߪଐὼߣߒߡ㜞ߊߥ޿ 8)‫ޕ‬ᧄ⎇ⓥߢߪ‫ߩࠤ࠲ࠗࠪޔ‬ኂ⩶ߢ޽ࠆਇቢో⩶Trichoderma harzianum ߦ⌕⋡ߒ‫ࡦ࠴ࠠޔ‬ಽ⸃㉂⚛

⓸⚛Ḯߣߒߡࡐ࡝ࡍࡊ࠻ࡦࠍ฽߻ၭ࿾ߢ
ߩၭ㙃 ߩ↥↢᧦ઙߦߟ޿ߡᬌ⸛ࠍട߃ߚ‫ࡦ࠴ࠠޔߚ߹ޕ‬὇⚛Ḯߣߒߡࠠ࠴ࡦ㧔㕖᥏ൻ‫ᧃ☳ޔ‬㧕޽ࠆ޿ߪߩಽ⸃ല₸ࠍ਄ߍࠆߚ߼ߦ‫ߚ޿↪ࠍ࡝ࠞ࡞ࠕޔ‬೨㧰-ࠣ࡞ࠦ࡯ࠬࠍ⛘ੇ㊀㊂ߣߒߡ 0.50 g‫ࡊࡍ࡝ࡐޔ‬ಣℂߦࠃࠅᓧࠄࠇߚ㕖᥏ൻࠠ࠴ࡦߣࠠ࠴ࡦಽ⸃

࠻ࡦ㧔0.20 g㧕‫ޔ‬㉂Უࠛࠠࠬ㧔0.01 g㧕
‫ޔ‬K2HPO4㧔0.07㉂⚛ߣߩ෻ᔕߦߟ޿ߡᬌ⸛ࠍട߃ߚ‫ޕ‬‫ޔ‬MgSO㧠࡮7H2O㧔0.05 g㧕ࠍg㧕‫ޔ‬KH2PO4㧔0.03 g㧕
฽߻ో㊂ 100 mL ߩၭ࿾ߦ‫ޔ‬೨ၭ㙃ߒߡ߅޿ߚ⩶ ታ㛎ⵝ⟎෸߮ታ㛎ᣇᴺ૕ࠍࠦ࡞ࠢࡏ࡯࡜࡯㧔Ǿ㧝cm㧕ߢᛂߜᛮ޿ߚ߽ߩ ૶↪⩶ᩣࠍኙᄤߏߣ㧝 ធ⒳ߒ‫ޔ‬120 rpm, 30͠ߢ࿁ォߒߥ

ଏ⹜⩶ᩣߣߒߡਇቢో⩶ Trichoderma harzianum㧔⩶⭇⎇ⓥᚲ TMICC60622㧕ࠍ↪޿ߚ‫ޕ‬߇ࠄၭ㙃ࠍⴕߞߚ‫ޕ‬ၭ㙃ᶧ߆ࠄ⚻ᤨ⊛ߦ৻ㇱࠍណขߒ‫(࡯࠲࡞ࠖࡈࡦ࡜ࡉࡦࡔޔ‬0.45 Ǵm㧕ࠍ↪޿ߡࠈㆊߒߚ߽ߩࠍၭ㙃ࠈᶧߣߒ‫ޔ‬㉂⚛ᵴᕈ᷹ቯߦ↪

 㕖᥏ൻࠠ࠴ࡦߩ⺞⵾⛘ੇ㊀㊂ߢ 5.0 g ߩࠠ࠴ࡦ TC-L㧔ਃᩕᎿᬺ㧔ᩣ㧕

޿ߚ‫ޕ‬


⵾㧕ࠍ 125 g ߩ 40%NaOH ᳓ṁᶧߦ Ộߐߖ‫ޔ‬ቶ⓸⚛Ḯߣߒߡ⎫㉄ࠕࡦࡕ࠾࠙ࡓࠍ฽߻ၭ᷷ߢᤨ‫ޘ‬ᡬᜈߒߥ߇ࠄ 12 ᤨ㑆᡼⟎ߒߚᓟ‫ޔ‬
⎈᳖ࠍ

࿾ߢߩၭ㙃

375 g ട߃‫ޔ‬ቢోߦṁ⸃ߒቶ᷷ߦᚯࠆ߹ߢࠁߞߊࠅ ࠠ࠴ࡦ㧔㕖᥏ൻ‫ޔᧃ☳ޔ‬ᓸ☳ᧃ 14 Ǵm 㧕ࠍ⛘ߣᠣᜈߒߚ‫ ߡ޿↪ࠍ㉄⎫ޔߡ޿⛯ޕ‬pH ࠍ 8.4 ߦ⺞g㧕‫ޔ‬KH2PO4
ੇ㊀㊂ߣߒߡ 0.50 g ‫(ޔ‬NH4)2SO㧔0.424ᢛߒߚ‫↢ޕ‬ᚑߒߚࠥ࡞ࠍࠈ೎ᓟ‫⇐⫳ޔ‬᳓ࠍ↪޿ߡ

㧔0.20 g㧕‫ޔ‬NaH2PO4㧔0.69 g㧕
‫ޔ‬MgSO㧠࡮7H2O㧔0.03
̶2̶㠽ข⋵↥ᬺᛛⴚ࠮ࡦ࠲࡯‫⎇ޓ‬ⓥႎ๔ͳ  g㧕‫ޔ‬Tween 80㧔0.02 g㧕
‫ޔ‬FeSO4࡮7H2O㧔0.50 mg㧕‫ޔ‬ߚ‫ޔ߅ߥޕ‬㉂⚛ᵴᕈߩ㧝න૏ߪ㧝ಽ㑆޽ߚࠅ㧝‫ޔ‬ZnSO4࡮7H2O㧔0.14 mg㧕
‫ޔ‬
MnSO4࡮5H2O㧔0.25 mg㧕mol ߩ p-࠾࠻ࡠࡈࠚࡁ࡯࡞ࠍ↢ᚑߔࠆߩߦᔅⷐ

CaCl2࡮2H2O㧔0.12 mg㧕ࠍ฽߻ో㊂ 100 mL ߩၭߥ㉂⚛㊂ߣቯ⟵ߒߚ‫ޕ‬

࿾ߦ 2.3.2 ߣห᭽ߦᬀ⩶ߒ‫ޔ‬ၭ㙃ࠍⴕߞߚ‫ޕ‬ ࠠ࠴ࡦߩಽ⸃෻ᔕ ㉂⚛ᵴᕈߩ᷹ቯ ⛘ੇ㊂ߣߒߡ 20 mg ߩࠠ࠴ࡦ㧔㕖᥏ൻࠠ࠴ࡦ޽ ࠠ࠴࠽࡯࠯ᵴᕈࠆ޿ߪ☳ᧃ⁁ࠠ࠴ࡦ㧕‫☻ޔ‬㉂⚛ᶧ㧔ࠠ࠴࠽࡯࠯ᵴᕈࠠ࠴࠽࡯࠯ᵴᕈߪ‫ޔ‬ᵴᕈ᷹ቯߩၮ⾰ߣߒߡࠣ࡝ߣߒߡ 0.5 U ಽ‫ޔ‬Ǫ-N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯ࠦ࡯࡞ࠠ࠴ࡦࠍ↪޿ࠆ Imoto ࠄߩᣇᴺߦࠃࠅ᷹ቯ

࠯ᵴᕈߣߒߡ 1.5 U ಽ㧕ࠍ฽߻ో㊂㧡mL ߩ 80 mM

9㧕ߒߚ ‫ޕ‬0.1%ࠣ࡝ࠦ࡯࡞ࠠ࠴ࡦߩ 0.1 M ㈶㉄࠽࠻㈶㉄࠽࠻࡝࠙ࡓṁᶧ㧔pH6.0㧕ࠍ 37͠ߢᡬᜈߒߚ‫ޕ‬࡝࠙ࡓ✭ⴣᶧ㧔pH 6.0㧕1.0 mL ߦ☻㉂⚛ᶧ 0.2 mL෻ᔕ㐿ᆎᓟ‫ޔ‬24 ᤨ㑆ߩᤨὐߢห㊂ߩ㉂⚛ᶧࠍߐࠄࠍട߃‫ޔ‬37͠ߢ 10 ಽ㑆෻ᔕߐߖߚ‫ޕ‬෻ᔕᓟ‫ࠄ޽ޔ‬ߦㅊടߒߚ‫ߦ⊛ᤨ⚻ޕ‬෻ᔕᶧߩ৻ㇱࠍขࠅ‫ޔ‬HPLC

߆ߓ߼⺞⵾ߒߡ߅޿ߚ Schales ߩ⹜⮎㧔0.05%ࡈࠚ

ࠍ↪޿ߡ‫ ࡓ࡜ࠞޔ‬Shodex NH2P-50-4E‫ޔ‬ṁ಴ᶧࠕ࡝ࠪࠕࡦൻࠞ࡝࠙ࡓ 0.5 M ὇㉄࠽࠻࡝࠙ࡓ᳓ṁ‫ޔ‬ᵹㅦ㧝mL/ಽ‫ޔ‬࠮࠻࠾࠻࡝࡞/H2O㧩70/30㧔v/v㧕

ᶧ㧕2.0 mL ࠍട߃‫ޔ‬15 ಽ㑆ᾚᴣߒߚ‫ޕ‬᡼಄ᓟ‫ޔ‬420ࠞ࡜ࡓ᷷ᐲ 40͠ߢ␜Ꮕዮ᛬ᬌ಴ߦࠃࠅ෻ᔕ↢ᚑnm ߩๆశᐲࠍ᷹ቯߒߚ‫☻ޕ‬㉂⚛ᶧߩઍࠊࠅߦห‛ࠍಽᨆߒߚ‫↢ޔ߅ߥޕ‬ᚑ‛ߩหቯߪᮡຠߣߩ଻㊂ߩ⫳⇐᳓ࠍട߃ห᭽ߩ෻ᔕ࡮᷹ቯࠍⴕ޿ኻᾖታ

ᜬᤨ㑆ߩᲧセߦࠃࠅⴕߞߚ‫ޕ‬㛎ߣߒߚ‫ޕ‬ኻᾖታ㛎ߩๆశᐲ୯߆ࠄ㉂⚛෻ᔕߒߚ߽ߩߩๆశᐲ୯ࠍᏅߒᒁ޿ߚ୯ࠍ▚಴ߒ‫ߩߎޔ‬ๆ ࠣ࡜ࡓࠬࠤ࡯࡞ߢߩࠠ࠴ࡦߩಽ⸃෻ᔕశᐲ୯ߣ‫ ߼ߓ߆ࠄ޽ޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒ

300 mL ኈ࠽ࠬဳࡈ࡜ࠬࠦߦ⛘ੇ㊂ߣߒߡ㧝 gࡦࠍ↪޿ߡⴕߞߚห᭽ߩ෻ᔕ߆ࠄ૞ᚑߒߡ߅޿ߚߩ㕖᥏ൻࠠ࠴ࡦ‫☻ޔ‬㉂⚛ᶧ㧔ࠠ࠴࠽࡯࠯ᵴᕈߣߒ

ᬌ㊂✢ߣ߆ࠄ㉂⚛ᵴᕈࠍ⸘▚ߒߚ‫ޔ߅ߥޕ‬㉂⚛ᵴߡ 13 U‫ޔ‬Ǫ-N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ᵴᕈᕈߩ㧝න૏ߪ㧝ಽ㑆޽ߚࠅ㧝mol ߩ N-ࠕ࠮࠴࡞ߣߒߡ 68 U ಽ㧕ࠍ౉ࠇ‫⇐⫳ߦࠄߐޔ‬᳓ࠍട߃ߡో

-D-ࠣ࡞ࠦࠨࡒࡦࠍ↢ᚑߔࠆߩߦᔅⷐߥ㉂⚛㊂ߣ㊂ 100 mL ߣߒߚ‫ޕ‬෻ᔕᶧߩ pH ߪ⺞ᢛߒߥ߆ߞߚ‫ޕ‬ቯ⟵ߒߚ‫ޕ‬37͠ߩ᳓ᶎਛߢᠣᜈࠍⴕ޿‫ߦ⊛ᤨ⚻ޔ‬෻ᔕᶧߩ৻ㇱࠍขࠅ‫ޔ‬HPLC ࠍ↪޿ߡ 2.5 ߣห᭽ߦ෻ᔕ↢ᚑ N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ᵴᕈ‛ࠍಽᨆߒߚ‫ޔ߅ߥޕ‬෻ᔕ㐿ᆎᓟ 24 ᤨ㑆⋡ߦඨಽǪ-N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ᵴᕈߪ‫ޔ‬p-࠾㊂ߩ㉂⚛ᶧࠍㅊടߒߚ‫ޕ‬෻ᔕ㐿ᆎ 48 ᤨ㑆ᓟ‫ޔ‬෻ᔕ࠻ࡠࡈࠚ࠾࡞㧞-ࠕ࠮࠻ࠕࡒ࠼-㧞-࠺ࠝࠠࠪ-Ǫ-D-

ᶧࠍ 15 ಽ㑆ᾚᴣߒ㉂⚛ࠍᄬᵴߐߖߚ‫ޕ‬ᰴ޿ߢ‫ޔ‬㆙ࠣ࡞ࠦࡇ࡜ࡁࠪ࠼㧔pNP-GlcAc㧕ࠍၮ⾰ߣߒߡ᷹ᔃಽ㔌ࠍⴕ޿㧔10,200㨓‫ޔ‬20 ಽ㧕‫ޔ‬਄Ẵࠍ㓸߼‫ޔ‬

ቯߒߚ 10)‫ޕ‬శ〝㐳㧝 cm ߩ⍹⧷࠮࡞ߦ 0.04 mL ߩᱷᷲߦ⫳⇐᳓ࠍട߃‫ޔ‬หߓ᧦ઙߢౣᐲ㆙ᔃಽ㔌ࠍ

5 mM pNP-GlcNAc ᳓ṁᶧ‫ޔ‬0.2 mL ߩ 1 M ㈶㉄࠽ⴕߞߚ‫ޕ‬਄Ẵᶧࠍวࠊߖߡ㒢ᄖࠈㆊࠍⴕ޿㧔ࠕ࠼࠻࡝࠙ࡓ㧔pH6.0㧕᳓ṁᶧ߅ࠃ߮☻㉂⚛ᶧࠍᦨ⚳ኈ‫ޔ‬ᓧࠄࡃࡦ࠹࠶ࠢ UK-10‫ޔ‬ಽሶ㊂ࠞ࠶࠻㧦10,000㧕

㊂ 2.0 mL ߦߥࠆࠃ߁ߦ౉ࠇ‫ߦ⊛ᤨ⚻ޔ‬෻ᔕ↢ᚑ‛ࠇߚࠈᶧࠍߐࠄߦಓ⚿ੇ῎ߒߚ‫ޕ‬ᓧࠄࠇߚ☳ᧃߦߢ޽ࠆ p-࠾࠻ࡠࡈࠚࡁ࡯࡞ߩᦨᄢๆ෼ᵄ㐳ߢ޽ࠆࡔ࠲ࡁ࡯࡞ࠍട߃‫ޔ‬ਇṁಽࠍ㒰෰ᓟ‫ޔ‬ᷫ࿶Ớ❗ߦ

337 nm ߩๆశᐲߩჇടࠍ⚻ᤨ⊛ߦ᷹ቯߒߚ‫ߩߎޕ‬ࠃࠅ↢ᚑߒߚᴉᲚࠍ಄ࡔ࠲ࡁ࡯࡞ߢᵞᵺߒߥ߇ࠄᵄ㐳ߦ߅ߌࠆಽሶๆశଥᢙ 3,500 M cm ࠍ↪޿ࠈ೎ߒߚ‫ޕ‬-1-1ߡ෻ᔕ↢ᚑ‛ߩࡕ࡞ᢙࠍ⸘▚ߒ‫ޔ‬㉂⚛ᵴᕈࠍ᳞߼
̶3̶ ⚿ᨐၭ࿾ߢၭ㙃ࠍⴕ޿‫ࡦ࠴ࠠޔ‬ಽ⸃㉂⚛ᵴᕈߩ⚻ᤨᄌ 6JCT\KCPWO ߦࠃࠆࠠ࠴ࡦಽ⸃㉂⚛ߩ↥↢ൻࠍ⺞ߴߚ⚿ᨐࠍ࿑㧞ߦ␜ߔ‫ޕ‬Ყセߩߚ߼ߦන৻ࠠ࠴ࡦࠍቢోߦಽ⸃ߒߡන♧ߢ޽ࠆ N-ࠕ࠮࠴὇⚛Ḯߣߒߡࠣ࡞ࠦ࡯ࠬࠍ↪޿ߚ⚿ᨐߦߟ޿ߡ߽

࡞-D-ࠣ࡞ࠦࠨࡒࡦࠍᓧࠆߚ߼ߦߪ‫ࡦ࠴ࠠޔ‬㎮ࠍ࡜ห᭽ߦ␜ߒߚ‫⥝ޕ‬๧޽ࠆߎߣߦ‫ޔ‬㕖᥏ൻࠠ࠴ࡦࠍࡦ࠳ࡓߦട᳓ಽ⸃ߔࠆࠛࡦ࠼ဳߩࠠ࠴࠽࡯࠯ߣ޽

ട߃ߚ႐วߦǪ-N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ࠆ⒟ᐲಽ⸃ߐࠇߚࠠ࠴ࡦಽሶ㎮ࠍᧃ┵߆ࠄട᳓ಽ

ᵴᕈߛߌ߇ⷰኤߐࠇߚ‫ޕ‬⸃ߒߡ޿ߊࠛࠠ࠰ဳߩǪ-N-ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ200 A࠾࠳࡯࠯ߩ૞↪߇ᔅⷐߣߥࠆ‫ޔߢߎߘޕ‬ਔ㉂⚛߇150㜞ᵴᕈߢ↥↢ߐࠇࠆ᧦ઙࠍᬌ⸛ߔࠆߎߣߣߒߚ‫ޕ‬100
Activity (U/flask)

T. harzianum ߩࠠ࠴ࡦಽ⸃㉂⚛ߪߔߢߦಽ㔌࡮♖⵾ߐࠇ⻉ᕈ⾰ߦߟ޿ߡႎ๔߇ߥߐࠇߡ޿ࠆ߇ 11)‫ࠃޔ‬ࠅ㜞޿↥↢᧦ઙࠍ⷗޿ߛߔߚ߼ߦ‫ޔ‬ၭ㙃᧦ઙߩᬌ⸛ࠍⴕߞߚ‫ޕ‬

50
0B150
10050

Intensity (cps)

002

468

10

Incubation period (day)AB0 10 20 30 40 50 60 70 80 90
2T (º)࿑㧞 ⓸⚛Ḯߣߒߡࡐ࡝ࡍࡊ࠻ࡦ‫ޔ‬὇⚛Ḯߣߒߡ &ࠣ࡞ࠦ࡯ࠬ ٠ ☳ᧃ⁁ࠠ࠴ࡦ ‫ߪߚ߹ ع‬㕖᥏ൻࠠ࠴ࡦ ٌ ࠍട߃ߚၭ࿾ߢ 6JCT\KCPWOߦࠃࠅ↥↢ߐࠇߚࠠ࠴࠽࡯࠯㧔#㧕߅ࠃ߮Ǫ0ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯㧔$㧕ᵴᕈߩ⚻ᤨᄌൻ࿑㧝☳ᧃ⁁ࠠ࠴ࡦ㧔#㧕߅ࠃ߮㕖᥏ൻࠠ࠴ࡦ㧔$㧕ߩ ߎߩၭ㙃᧦ઙߢߪ‫ⷰ߇↢↥ߩ࠯࡯࠽࠴ࠠޔ‬ኤߐ: ✢࿁᛬ᦛ✢ࠇߥ߆ߞߚߚ߼ߦ‫ޔ‬ᰴߦ὇⚛Ḯߣߒߡࠠ࠴ࡦࠍട ᣢႎߢߪ‫ޔ‬὇⚛Ḯߣߒߡ☳ᧃ⁁ߩࠠ࠴ࡦࠍᷝട߃ߚᤨߩ⓸⚛Ḯߩᓇ㗀ߦߟ޿ߡ⺞ߴࠆߎߣߦߒߚ‫ޕ‬ߒߡ޿ࠆ႐ว߇ᄙ޿߇‫ߪࡦ࠴ࠠޔ‬ᒝ࿕ߥ⚿᥏᭴ㅧ⓸⚛Ḯߣߒߡߪ‫ޕߚ޿↪ࠍࡓ࠙࠾ࡕࡦࠕ㉄⎫ޔ‬὇ࠍ᦭ߔࠆߚ߼ߦ‫ߩࡦ࠴ࠠޔ‬ᕈ⁁߇㉂⚛ߩ↢↥ᕈߦ⚛Ḯߣߒߡട߃ࠆࠠ࠴ࡦߩᕈ⁁ߩ㆑޿ߦߟ޿ߡ߽ᓇ㗀ࠍਈ߃ࠆߎߣ߇੍ᗐߐࠇࠆ‫࡞ࠕޔ߼ߚߩߘޕ‬⍮⷗ࠍᓧࠆߚ߼ߦ‫ޔࡦ࠴ࠠߩ⁁ᧃ☳ޔ‬㕖᥏ൻಣℂ

ࠞ࡝ߢ⤘Ảಣℂࠍⴕߞߚ㕖᥏ൻࠠ࠴ࡦߦߟ޿ߡ߽ࠍߒߚࠠ࠴ࡦ‫ޔ‬ᓸ☳⎈ൻߒߚࠠ࠴ࡦ㧔14m㧕ࠍߘᬌ⸛ࠍട߃ߚ‫ޕ‬㉂⚛ߩ↢↥ᕈࠍᬌ⸛ߔࠆߦవ┙ߞࠇߙࠇട߃ߚ᧦ઙߢၭ㙃ࠍⴕߞߚ‫⚿ߩߘޕ‬ᨐ‫⓸ޔ‬ߡ‫ߢ࡝ࠞ࡞ࠕޔ‬ಣℂߐࠇߚࠠ࠴ࡦߩ X ✢࿁᛬ታ㛎⚛Ḯߣߒߡࡐ࡝ࡍࡊ࠻ࡦࠍട߃ߚ႐วߣ⇣ߥࠅ‫ޔ‬ࠍⴕ޿‫ߦࡦ࠴ࠠߩ⁁ᧃ☳ޔ‬Ყセߒߡ‫ޔ‬᣿⍎ߥࡇ࡯ਔ㉂⚛ߩᵴᕈ߇ⷰኤߐࠇ‫ߢࡦ࠴ࠠߩ⁁ᧃ☳ޔ‬ᵴᕈࠢߩߥ޿ߎߣࠍ⏕⹺ߒߡ߅޿ߚ㧔࿑㧝㧕‫ࠠޔߚ߹ޕ‬߇㜞޿⚿ᨐߣߥߞߚ‫ޔߒߛߚޕ‬㉂⚛↥↢ߦⷐߔࠆ࠴ࡦߪࠕ࡞ࠞ࡝⤘Ảಣℂᓟ‫ޔ‬ቯ㊂⊛ߦ࿁෼ߔࠆߎᣣᢙߪࡐ࡝ࡍࡊ࠻ࡦߩ႐วࠃࠅ߽㐳ߊߥߞߚ‫ޕ‬὇ߣ߇ߢ߈ߚ‫ޕ‬⚛Ḯߣߒߡ↪޿ࠆࠠ࠴ࡦߩᒻ⁁ߩ㆑޿ߦ᣿⍎ߥᏅߎߩ㕖᥏ൻࠠ࠴ࡦ߅ࠃ߮☳ᧃ⁁ࠠ࠴ࡦࠍන৻὇⚛Ḯߣߒ‫⚛⓸ߦࠄߐޔ‬Ḯߣߒߡࡍࡊ࠻ࡦࠍട߃ߚߪ⹺߼ࠄࠇߕ‫⚿ޔ‬᥏ൻᐲ߇ૐߊߡ߽㉂⚛↢↥ᕈߩᡷༀߪ⷗ࠄࠇߥ߆ߞߚ㧔࿑㧟㧕‫ޕ‬̶4̶㠽ข⋵↥ᬺᛛⴚ࠮ࡦ࠲࡯‫⎇ޓ‬ⓥႎ๔ͳ  㧠ߦ␜ߔ‫ޕ‬50෻ᔕᤨ㑆ߩ⚻ㆊߣߣ߽ߦ෼㊂ߪჇടߒ‫ޔ‬㕖᥏⾰A40ࠠ࠴ࡦࠍ಴⊒‛⾰ߣߒߚ႐วߪ‫ޔ‬෻ᔕ㐿ᆎᓟ 30ᤨ㑆⋡ߩ෼㊂߇߶߷ 㧑ߢ޽ࠅ‫ߡߴߔࡦ࠴ࠠޔ‬߇ Nࠕ࠮࠴࡞Dࠣ࡞ࠦࠨࡒࡦߦᄌ឵ߐࠇߡ޿

10ࠆߎߣ߇ಽ߆ߞߚ‫৻ޕ‬ᣇ‫ࠍࡦ࠴ࠠ⁁ᧃ☳ޔ‬಴⊒‛

0200

⾰ߣߒߚ႐วߪߘߩ↢ᚑ߇߶ߣࠎߤⷰኤߐࠇߥ߆

Bߞߚ‫ޕ‬

150
100

100

50

80

00

105
Incubation period (day)Yield (%)

Activity (U/flask)

20156040࿑㧟 ⓸⚛Ḯߣߒߡ⎫㉄ࠕࡦࡕ࠾࠙ࡓ‫ޔ‬὇⚛Ḯߣߒ

20ߡ☳ᧃ⁁ࠠ࠴ࡦ ً 㕖᥏ൻࠠ࠴ࡦ ‫ߚ߹ غ‬

00ߪ ᓸ ☳ ᧃ ൻ ࠠ ࠴ ࡦ  ٨ ࠍ ട ߃ ߚ ၭ ࿾ ߢ 6

1020304050

Reaction time (hrs)

JCT\KCPWOߦࠃࠅ↥↢ߐࠇߚࠠ࠴࠽࡯࠯㧔#㧕߅ࠃ߮࿑  6JCT\KCPWO ☻㉂⚛ᶧߣ㕖᥏ൻࠠ࠴ࡦ ٨Ǫ0ࠕ࠮࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯㧔$㧕ᵴᕈߩ⚻ᤨ߹ߚߪ☳ᧃ⁁ࠠ࠴ࡦ ً ߣߩ෻ᔕߦࠃࠆ 0ࠕ࠮࠴ᄌൻ࡞&ࠣ࡞ࠦࠨࡒࡦߩ↢ᚑ㧚⍫ශߩᤨὐߢ㉂⚛ࠍㅊടߒߚ㧚 ၭ㙃ࠈᶧߦࠃࠆࠠ࠴ࡦߩಽ⸃෻ᔕ  ෻ᔕߩ⚻ᤨᄌൻ ࠬࠤ࡯࡞ࠕ࠶ࡊታ㛎

3.1 ߢㅀߴߚ‫ޔ‬὇⚛Ḯߣߒߡ☳ᧃ⁁ࠠ࠴ࡦ‫⚛⓸ޔ‬ ⛘ੇ㊀㊂ߣߒߡ㧝㨓ࠣ࡜ࡓߩ㕖᥏ൻࠠ࠴ࡦࠍ಴Ḯߣߒߡ⎫㉄ࠕࡦࡕ࠾࠙ࡓࠍട߃ߡၭ㙃ࠍⴕߞߚ⊒ේᢱߣߒߡ෻ᔕࠍⴕ޿‫↢ޔ‬ᚑߒߚ N-ࠕ࠮࠴࡞႐ว‫ޔ‬ၭ㙃ࠈᶧਛߦߪࠠ࠴࠽࡯࠯߅ࠃ߮Ǫ-N-ࠕ࠮

-D-ࠣ࡞ࠦࠨࡒࡦ㊂ࠍ HPLC ߢಽᨆߒߚ⚿ᨐ‫ޔ‬࠴࡞ࡋࠠ࠰ࠨࡒ࠾࠳࡯࠯ߩᵴᕈߪⷰኤߐࠇࠆ߽ߩᤨ㑆ߩ෻ᔕߢ 㧑ߩ෼㊂ߢ޽ߞߚ㧔࠺࡯࠲⋭⇛㧕‫ޕ‬ߩ‫ޔ‬HPLC ߦࠃࠆၭ㙃ࠈᶧߩಽᨆߢߪ‫ޔ‬N-ࠕ࠮࠴ߎࠇߪ‫ޔ‬3.2.1 ߦ߅ߌࠆዊࠬࠤ࡯࡞ߢߩታ㛎ߩ႐ว

࡞-D-ࠣ࡞ࠦࠨࡒࡦߩሽ࿷ߪⷰኤߐࠇߥ߆ߞߚ㧔࠺ࠃࠅૐ޿୯ߢ޽ߞߚ‫ߩߎޔߚ߹ޕ‬෻ᔕᶧ߆ࠄ N-࡯࠲⋭⇛㧕‫↢ޔߪߣߎߩߎޕ‬ᚑߒߚ N-ࠕ࠮࠴࡞-D-

ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦࠍಽ㔌࡮♖⵾ߒߚߣߎࠈ‫ޔ‬ࠣ࡞ࠦࠨࡒࡦ߇⩶૕ߦࠃࠅઍ⻢ߐࠇߡ޿ࠆߣផቯ

ࠠ࠴ࡦߩ㊀㊂ߦኻߒߡ 58%ߢ࿁෼ߐࠇߚ‫ ޕ‬ߐࠇࠆ‫࡯࠲࡞ࠖࡈࡦ࡜ࡉࡦࡔޔߢߎߘޕ‬㧔0.45Ǵm㧕ࠍ↪޿ߡၭ㙃ᶧ߆ࠄ⩶૕ࠍ㒰෰ߒ‫ޔ‬ᓧࠄࠇߚ㧠㧚⠨ኤၭ㙃ࠈᶧࠍ☻㉂⚛ᶧߣߒ‫࠴ࠠߡ߼ߚࠄ޽ࠍࠇߎޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦࠍᓧࠆߚ߼ߩᦨ߽

ࡦߣ෻ᔕߐߖ‫ޔ‬N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩ↢ᚑ

◲නߥᣇᴺߪ㉄ട᳓ಽ⸃ߢ޽ࠆ߇‫ߩߎޔ‬ᣇᴺߢߪߦߟ޿ߡᬌ⸛ߔࠆߎߣߣߒߚ‫ౕޕ‬૕⊛ߦߪ‫ޔ‬㕖᥏⣕ࠕ࠮࠴࡞ൻ෻ᔕ߇ઃ㓐ߔࠆߚ߼ߦ‫࡞࠴࠮ࠕޔ‬ၮൻߒߚࠠ࠴ࡦߣ☳ᧃ⁁ࠠ࠴ࡦߘࠇߙࠇࠍၮ⾰ߣߒߩ⣕㔌ࠍᛥ߃ࠆߚ߼ߩ᧦ઙ⸳ቯ߇ᔅⷐߣߥࠆ‫ޕ‬ߡ☻㉂⚛ᶧߣߩ෻ᔕࠍⴕ޿‫ࡦ࡝ࡊࡦࠨߦ⊛ᤨ⚻ޔ‬40͠‫ޔ‬12 ᤨ㑆ߩႮ㉄ട᳓ಽ⸃෻ᔕ㧔200g ߩࠠ࠴ࡦࠣߒߡ N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦ㊂ࠍ᳞߼‫ࠠޔ‬ߦኻߒߡ 1kg ߩỚႮ㉄㧕ߢ෼㊂⚂ 60㧑ߩႎ๔߇޽

࠴ࡦ㊀㊂ߦኻߔࠆ෼₸ࠍ⸘▚ߒߚ‫⚿ߩߘޕ‬ᨐࠍ࿑ࠆ߇‫ޔ‬ଐὼߣߒߡ 10%⒟ᐲߩੑ㊂૕߿ᢙ%⒟ᐲߩ
̶5̶ࠝ࡝ࠧࡑ࡯߇ᱷሽߒߡ޿ࠆ 6a)‫ޕ‬㉂⚛ࠍ↪޿ߚಽ⸃ᧄ⎇ⓥߢߪ‫ޔ‬㜞⚿᥏ᕈߩࠞ࠾Ზ↱᧪ߩǩ-ࠠ࠴ࡦ෻ᔕߣߒߡ‫ޔ‬Nocardia orientalis ߩ࿕ቯൻ㉂⚛ࠍ↪

ߩࠕ࡞ࠞ࡝⤘Ảಣℂࠍⴕ޿‫ࠍࡦ࠴ࠠߩߎޔ‬὇⚛Ḯ޿ߚࡃࠗࠝ࡝ࠕࠢ࠲࡯ਛߢ‫⵾⺞ߢ㉄߼ߓ߆ࠄ޽ޔ‬ߣߒߚᤨߩ T. harzianum ߦࠃࠆࠠ࠴ࡦಽ⸃㉂⚛ߣߒߡ߅޿ߚࠠ࠴ࡦࠝ࡝ࠧ♧ߩಽ⸃ࠍⴕ޿‫ޔ‬ㅪ⛯ߒ

N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩ↢ᚑߦߟ޿ߡᬌ⸛

ߡ130ᣣ㑆ߦࠊߚࠅ90㧑એ਄ߩ෼₸ߢN-ࠕ࠮࠴࡞ࠍട߃ߚ‫ᧄޕ‬ታ㛎ߩ⚿ᨐ߆ࠄ‫ޔ‬T. harzianum ߩࠠ࠴-D-ࠣ࡞ࠦࠨࡒࡦࠍᓧߡ޿ࠆ߇ 8a)‫ޔ‬㉂⚛ಽ⸃ߩၮ⾰ࡦಽ⸃㉂⚛ᵴᕈߪ‫⚛⓸ޔ‬Ḯߩᓇ㗀ࠍฃߌࠆߎߣ߇ߣߒߡ↪޿ࠆࠠ࠴ࡦࠝ࡝ࠧ♧ߩ⺞⵾Ბ㓏ߢߩ಴⊒᣿ࠄ߆ߣߥࠅ‫ޔ‬⎫㉄ࠕࡦࡕ࠾࠙ࡓ߇⦟ᅢߢ޽ߞߚ‫ޕ‬ේᢱߩࠠ࠴ࡦߦኻߔࠆ෼㊂߇ 40㧑ߢ޽ࠆߎߣࠍ߹ߚ‫ޔ‬὇⚛Ḯߣߒߡߪ㕖᥏ൻࠠ࠴ࡦࠍ↪޿ߚ႐ว⠨߃ࠆߣ‫ޔ‬಴⊒ේᢱߩࠠ࠴ࡦߦኻߔࠆᦨ⚳෼₸ߪߦᵴᕈ߇㜞ߊߥࠆߎߣ߇੍ᗐߐࠇߚ߇‫ߩࡦ࠴ࠠޔ‬㧟ഀ⒟ᐲߩ෼₸ߣផቯߐࠇࠆ‫ޕ‬⒳㘃㧔☳ᧃ‫ޔ‬㕖᥏ൻ‫ޔ‬ᓸ☳ᧃൻ㧕ߦࠃࠄߕ㉂⚛ᵴ㜞ಽሶ㊂ࠠ࠴ࡦࠍ಴⊒ේᢱߣߒߚ㉂⚛෻ᔕߦࠃᕈ߇ⷰኤߐࠇߚߚ߼‫⚿ߩࡦ࠴ࠠޔ‬᥏ᕈߪ․ߦ໧㗴ࠆಽ⸃߽⹜ߺࠄࠇߡ޿ࠆ‫ߩߎޕ‬႐ว‫⚿ߩࡦ࠴ࠠޔ‬ߣߥࠄߥ޿ߣ⠨߃ࠄࠇߚ‫ޕ‬ᒰೋߪ‫ޔ‬὇⚛Ḯߣߒߡ᥏ᕈ߇㜞޿ߣ㉂⚛ߣߩⷫ๺ᕈ߇ૐߊߥࠆߚ߼ߦ‫ޔ‬ട߃ߚࠠ࠴ࡦߩಽ⸃ߦࠃࠅ↢ᚑߒߚ N-ࠕ࠮࠴࡞ࠗࠞ↱᧪ߩǪ-ࠠ࠴ࡦߩ೑↪߇ᬌ⸛ߐࠇߚ‫ޕ‬Ꮢ⽼ߩ-D-ࠣ࡞ࠦࠨࡒࡦߩၭ㙃ࠈᶧਛߦ߅ߌࠆሽ࿷߇ᦼ

࠮࡞࡜࡯࠯߿࡝ࡄ࡯࠯ߥߤߩᄙ♧㘃ಽ⸃㉂⚛ߣǪ-ᓙߐࠇߚ߇‫੍ޔ‬ᗐߦ෻ߒߡߘߩሽ࿷߇ⷰኤߐࠇߥࠠ࠴ࡦߣߩ෻ᔕߢߪ‫ޔ‬Trichoderma viride ߩ࠮࡞࡜߆ߞߚߚ߼‫ޔ‬ၭ㙃ࠈᶧ߆ࠄߩ N-ࠕ࠮࠴࡞-D-ࠣ࡞

࡯࠯ߣߩ 37͠‫ޔ‬㧤ᣣ㑆ߩ෻ᔕߢ 76㧑ߩ෼㊂ߢ⋡⊛ࠦࠨࡒࡦߩಽ㔌࡮♖⵾ߪਇน⢻ߢ޽ߞߚ‫ࠄߘ߅ޕ‬7b㧕‫ޕ‬Ǫ-ࠠ࠴ࡦߩ Burkholderia cepaciaߊ‫⩶ޔ‬૕ߦࠃࠅ↢ᚑߒߚ N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨߩࠠ࠴ࡦಽ⸃㉂⚛ߦࠃࠆಽ⸃ߢߪ‫ޔ‬㧝ᣣߢ 85㧑એࡒࡦ߇ઍ⻢ߐࠇߚ߽ߩߣ⠨߃ࠄࠇߚ‫ޔ߼ߚߩߘޕ‬਄ߩ෼㊂ࠍႎ๔ߒߡ޿ࠆ ‫ޔࠄ߇ߥߒ߆ߒޕ‬ណ▚

৻ᣤ⩶૕ࠍ㒰෰ߒߚ㉂⚛ᶧࠍ࿁෼ߒ‫ߩߎޔ‬㉂⚛ᶧߩว߁ේᢱ⏕଻ࠍ⠨߃ࠆߣǪ-ࠠ࠴ࡦߩᎿᬺ⊛ߥߣࠠ࠴ࡦߣࠍౣᐲ෻ᔕߐߖࠆߎߣࠍᬌ⸛ߒߚߣߎ೑↪ߪ⃻ታ⊛ߢߪߥ޿ߣ⠨߃ࠄࠇࠆ‫ޕ‬ࠈ‫ޔ‬㜞෼₸ߢ N-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦࠍᓧࠆ‛ࠍᓧߡ޿ࠆ7c)৻ᣇ‫ࠄ߆ࡆࠛ߿࠾ࠞޔ‬᛽಴ߐࠇࠆǩ-ࠠ࠴ࡦߢߪ‫ޔ‬ߎߣ߇ߢ߈ߚ‫ߩߎޕ‬႐ว‫߽ࠅࠃࡦ࠴ࠠ⁁ᧃ☳ޔ‬㕖

Aeromonas hydrophila ߣߩ෻ᔕ߇ႎ๔ߐࠇߡ߅ࠅ‫ޔ‬᥏ൻࠠ࠴ࡦߩᣇ߇㉂⚛ߦࠃࠆಽ⸃ࠍฃߌ߿ߔߊ‫ޔ‬ࡈ࡟࡯ࠢ⁁ߩǩ-ࠠ࠴ࡦࠍේᢱߣߒߡ 10 ᣣ㑆ߢN-ࠕ࠮࠴࡞-D-ࠣ࡞ࠦࠨࡒࡦߩല₸⊛↢↥ᣇᴺߩ

‫ޔߚ߹ޕ‬೎ߩࠣ࡞ߚ߼ߩ೨ಣℂߣߒߡߪࠕ࡞ࠞ࡝⤘Ảಣℂߪ᦭ലߢ

77㧑ߩ෼㊂߇ႎ๔ߐࠇߡ޿ࠆ8b)

࡯ࡊߪ 85㧑࡝ࡦ㉄ߢ⤘Ảߐߖߚǩ-ࠠ࠴ࡦ߆ࠄ޽ࠆߎߣ߇ಽ߆ߞߚ‫ޕ‬

Aeromonas ዻ㧔⒳ߪᧂหቯ㧕ߩ☻㉂⚛ߦࠃࠅ㧡ᣣ߹ߚ‫ޔ‬෼₸ߩὐ߆ࠄ⷗ࠆߣ‫ᧄޔ‬ᴺߪࠕ࡞ࠞ࡝ಣ㑆ߢ 83㧑ߩ෼₸ࠍႎ๔ߒߡ޿ࠆ 8c)‫⤘㉄ࡦ࡝ޕ‬ẢࠠℂߩᲑ㓏ߢߩ෼㊂ૐਅ߇߶ߣࠎߤߥߊ㧔߶߷ቯ㊂

࠴ࡦߩ Trichoderma reesei ߅ࠃ߮ Cellulomonas sp.

⊛ߦ࿁෼ߐࠇࠆ㧕‫ޔ‬ߒ߆߽㉂⚛෻ᔕᤨߩ෼㊂߽㜞޿↱᧪ߩ㉂⚛ߦࠃࠆಽ⸃ߩႎ๔߽޽ࠆ߇‫ޔ‬෼㊂ߪߎߣ߆ࠄ㧔100㧑㧕‫⚿ޔ‬ᨐ⊛ߦ㜞޿ᦨ⚳෼₸߇⷗ㄟ

50㧑⒟ᐲߢ޽ࠆ 8d)‫ޕ‬߼ࠆ‫ࠆߔ↪૶ޔߚ߹ޕ‬ᓸ↢‛߿⮎ຠ߽․ᱶߥ߽ߩ෻ᔕല₸ࠍ਄ߍࠆߚ߼ߦǩ-෸߮Ǫ-ࠠ࠴ࡦߩᓸߢߥ޿ߎߣࠍ⠨߃ࠆߣታ↪ൻߦะߌߡߪ᦭ᦸߢ޽☳ ᧃ ൻ ࠍ ⴕ ޿ ‫ ޔ‬Burkholderia cepacia ‫ ޔ‬Bacillusࠆߣ⠨߃ࠄࠇࠆ‫ߚߒࡊ࠶ࠕ࡞࡯ࠤࠬޔߒߛߚޕ‬႐

licheniformis ↱᧪ߩࠠ࠴ࡦಽ⸃☻㉂⚛ᶧࠍ↪޿ߚวߩ෻ᔕᕈߩૐਅ‫ࡦ࠴ࠠߪߦࠄߐޔ‬ಽ⸃㉂⚛↢↥ߒ߆ߒߥ߇ࠄ‫ޔ‬⚿ᨐߦߟ޿ߡ߽ႎ๔ߐࠇߡ޿ࠆ ‫ޕ‬ߩߚ߼ߩ T. hartianum ߩၭ㙃ᤨ㑆߇㐳޿ߎߣ߇໧

ࠠ࠴ࡦߩᓸ☳ᧃൻߦߪᄙߊߩࠛࡀ࡞ࠡ࡯߇ᔅⷐߢ

㗴ߢ޽ࠆߚ߼ߦ‫੹ޔ‬ᓟߪߎߩὐߦߟ޿ߡᬌ⸛ࠍߔ

޽ࠅ‫߽࠻ࠬࠦޔ‬㜞ߊઃߊߎߣ߇໧㗴ߣߥࠆ‫ޕ‬ࠆᔅⷐ߇޽ࠆ‫ޕ‬8e)̶6̶㠽ข⋵↥ᬺᛛⴚ࠮ࡦ࠲࡯‫⎇ޓ‬ⓥႎ๔ͳ  A. Nakayama, E. Muraki, S. Aiba; Chem. Lett., 30,㧡㧚߅ࠊࠅߦp. 308-309 (2001).

㉂⚛↢↥ߩߚ߼ߩၭ㙃ᤨ㑆ߩ⍴❗ߦߟ޿ߡ‫ߐޔ‬8) a) K. Sakai, T. Uchiyama, Y. Matahira, F. Nanjo; J.

ࠄߦᬌ⸛ߔࠆᔅⷐ߇޽ࠆ‫ࡊ࠶ࠕ࡞࡯ࠤࠬޔߚ߹ޕ‬Ferment. Bioeng., 72, p. 168-172 (1991), b) H.

ߦ઻߁෼㊂ߩૐਅ߇⷗ࠄࠇࠆߚ߼ߦ‫ޔ‬෻ᔕല₸ࠍ

Sashiwa, S. Fijishima, N. Yamano, N. Kawasaki, A.

਄ߍࠆߚ߼ߩᎿᄦ߇ᔅⷐߢ޽ࠆߣ⠨߃ࠄࠇࠆ‫ߐޕ‬

Nakayama, E. Muraki, K. Hiraga, K. Oda, S. Aiba;ࠄߦ‫ߩࡦ࠴ࠠޔ‬ಽ⸃ߪⴕ߁߇ N-ࠕ࠮࠴࡞-D-ࠣ࡞

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ࠦࠨࡒࡦࠍઍ⻢ߒߥ޿ᓸ↢‛ࠍ↪޿ࠆߎߣ߇ߢ߈

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ࠇ߫‫ࠅࠃޔ‬ല₸⊛ߥ↢↥߇ᦼᓙߐࠇࠆ‫ޕ‬

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ᧄ⎇ⓥߩߚ߼ߦ‫ޔ‬T. harzianum㧔TMICC60622㧕

Kuttiyawong, M. Sukwattanasinitt; Carbohydr.

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̶7̶