Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ISSN 1348-5474

Dec. 2012╙Ꮞ╙ภ

Journal of Japanese Society for ExtremophilesᏎ㗡⸒㩷 ………………………………………………………… 46⎇ⓥᦨ೨✢㩷 …………………………………………………… 47
ORIGINAL PAPER㩷 …………………………………………

49

MINI REVIEW㩷 ………………………………………………

64ቇળ⸥੐㩷 ……………………………………………………… 105
ቇળ⹹ᛩⓂⷙቯ䊶ၫ╩ⷐ㗔㩷 …………………………………

108ળೣ䊶⚦ೣ㩷 ……………………………………………………

110ㆇ༡૕೙㩷 ……………………………………………………… 112

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Ꮞ㗡⸒⹏ଔߦߟ޿ߡ ᰴߦ⹏ଔߣ޿߁㕙߆ࠄߺߚࠨࠗࠛࡦࠬߣ࠹ࠢࡁࡠ
ࠫ࡯ߦߟ޿ߡ‫⎇ޕ‬ⓥ⠪ߩਛߦߪ‫ޔ‬ᓔ‫ߡߒߦޘ‬቟ᤃߦࠨࠗࠛࡦࠬߣ࠹ࠢࡁࡠࠫࠍ❬ߍߡ⸒߁ੱ߇޿߹ߔ‫ޕ‬ࠨࠗࠛࡦࠬߢߪ⁛ࠅࠃ߇ࠅߢ߽⦟޿ߩߢߔ߇‫ࠢ࠹ޔ‬ࡁࡠࠫ࡯ߢߪߘ߁ߪ޿߈߹ߖࠎ‫ߪߣ࡯ࠫࡠࡁࠢ࠹ޕ‬޽ߊ߹ߢઁ⠪ߣߩᲧセ⹏ଔߦ߅޿ߡ‫⥄ޔ‬ಽߩ⷗ߟߌ
ߚ‫ޔ‬㐿⊒ߒߚ‫ޔ‬ᚻᴺ߿‛ߦ૗ࠄ߆ߩࠕ࠼ࡃࡦ࠹࡯ࠫ
߇޽ࠆ႐วߦߩߺ⸒߃ࠆ⸒⪲ߢߔ‫ߩ࡯ࠫࡠࡁࠢ࠹ޕ‬ቴⷰ⊛⹏ଔᚻᲑߣߒߡߩ․⸵ߦ߅޿ߡߪ‫ޔ‬ᣂⷙᕈߣㅴᱠᕈ߇ࠕࡦ࠼᧦ઙߢߥߌࠇ߫ߥࠄߥ޿ߎߣ߆ࠄ߽
ಽ߆ࠅ߹ߔ‫ޕ‬ ᚒ߇࿖ߪ 1000 ళ౞ߦ߽㆐ߒࠃ߁ߣߔࠆ⿥୫㊄ᄢ࿖ߢߔ‫߿߹޿ޕ‬࿖᳃ో૕ߢߤ߁ߒߚࠄή㚝㆜޿ࠍᱛ߼ࠄࠇࠆ߆⌀೶ߦ⠨߃ࠆߴ߈ߢߔ‫⎇ߩߜߎߜ޽ޕ‬ⓥ
ᯏ㑐ߢหߓࠃ߁ߥ⎇ⓥ߇ⴕࠊࠇ‫ౕ߆ߥ߆ߥޔ‬૕⊛ᚑᨐߦ⚿߮ߟ߆ߥ޿⃻ታߦෂᗋࠍⷡ߃߹ߔ‫ߩߎߗߥޕ‬ࠃ߁ߥ⁁ᴫ߇↢߹ࠇߡ޿ࠆ߆ߣ޿߁ℂ↱ߩ৻ߟ߇⎇ⓥᚑᨐߩ⹏ଔߦ޽ࠆߣ⠨߃߹ߔ‫߁߽ޕ‬ዋߒ‫ޔ‬ᚑᨐࠍቴⷰ⊛ߦ‫౏ޔ‬ᐔߦᱜ⏕ߦ⹏ଔߒߡ‫ޔ‬ല₸⊛ߥ⎇ⓥ⾗
㊄ߩ૶޿ᣇࠍ⠨߃ࠆߴ߈ߢߔ‫ߩߘޕ‬ὑߦ৻⇟㊀ⷐߥߎߣߪߘࠇߛߌߩ⹏ଔ⢻ജࠍ᦭ߔࠆੱߩሽ࿷ߛߣᕁ
޿߹ߔ‫ᧄޔࠅࠃ߁޿ߣޕ‬ᒰߩߣߎࠈߪ‫⹏ޔ‬ଔߩዪ㕙ߢ‫ޔ‬ᕁߞߚߎߣࠍߪߞ߈ࠅ⸒߁ੱߩሽ࿷ߛߣᕁ޿߹
ߔ‫⎇ޕ‬ⓥ⠪ߩ㓸߹ࠅߪනߥࠆᖱႎ੤឵ߩ႐ߢߪߥߊ‫ޔ‬߅੕޿ߩ⎇ⓥ࠹࡯ࡑߩᗧ⟵‫⎇ޔ‬ⓥᚑᨐߦߟ޿ߡ₸⋥ߦ⹏ଔ‫ޔ‬ᛕ್ߒ߁ࠆ႐ߢߥߊߡߪㅴᱠߪή޿ߣᕁ޿
߹ߔ‫ޕ‬એ೨ߪቇળߥߤߢ‫ޔ‬ડᬺੱ߇ᄢቇ߆ࠄߩ⎇ⓥ⊒⴫ߦߟ޿ߡ߆ߥࠅㄆㄍߥ⾰໧ࠍߔࠆ႐㕙߽޽ߞߚࠃ߁ߦᕁ߁ߩߢߔ߇‫ᦨޔ‬ㄭߪ޽߹ࠅ⷗ฃߌ߹ߖࠎ‫ޕ‬ᄢቇߪᰴ਎ઍߩੱ᧚㙃ᚑߣ޿߁ࡒ࠶࡚ࠪࡦ߇޽ࠅ߹
ߔߩߢ೎ߦ⠨߃ࠆߣߒߡ‫ޔ‬࿖᳃ߩ⒢㊄ߢ⎇ⓥࠍⴕߞ
ߡ޿ࠆ⎇ⓥ⠪ߪ⥄ಽߩ⎇ⓥ࠹࡯ࡑߩᗧ⟵‫⎇ޔ‬ⓥᚑᨐ
߇ߤߩࠃ߁ߥᒻߢ↥ᬺߦᵴ߆ߖࠆ߆ࠍᏱߦᔨ㗡ߦ߅߈ߥ߇ࠄ઀੐ࠍߔࠆߴ߈ߢߔ‫ޕ‬ή⺰‫⥄⑳ޔ‬り߽⾌↪
ኻലᨐߩⷰὐ߆ࠄ߿ࠆߦ୯ߔࠆ⎇ⓥߢ޽ࠆ߆⥄ᚓߒߥ߇ࠄㅴ߼ߡ޿ࠆߟ߽ࠅߢߔ‫ޕ‬ ⑳ߪ㐳ᐕ᳃㑆ડᬺߢ⚻༡ߩ⃻ታࠍ⋡ߩᒰߚࠅߦߒ
ߡ᧪߹ߒߚߒ‫߽ࠄ⥄ޔ‬ᣂ੐ᬺഃᚑߩ㔍ߒߐࠍ૕㛎ߒߡ߈߹ߒߚ‫ޕ‬ડᬺ⚻༡ߦ߅ߌࠆ⹏ଔߩታ଀ߢ⸒߃߫‫ޔ‬ߘߩ෩ߒߐࠍታᗵߒߚߩߪ‫⸳ޔ‬஻ᛩ⾗᩺ઙߦߟ޿ߡ
ߩ⚻༡㒯ߩนุ⹏ଔߣ⚵❱ࡔࡦࡃ࡯ߩᬺ❣⹏ଔߢߔ‫ޕ‬޿ߕࠇ߽ડᬺሽ┙ࠍᏀฝߒ߆ߨߥ޿㊀ⷐߥ⹏ଔߢߒ
ߚ‫⹏ޕ‬ଔ߇↞޿ߣ‫ޔ‬ડᬺߪୟ↥ߩෂᯏࠍㄫ߃߹ߔߒ‫ޔ‬࿖ኅߢߐ߃߽‫⩨ޔ‬ᄢߥ୫㊄ߢ࿖᳃ߩ዁᧪ࠍෂ߁޿߽
ߩߦߒ߹ߔ‫ޕ‬ᱜ⏕ߢ౏ᐔߥ‛Ꮕߒߩሽ࿷ߣ‫ߎ޿૶ޔ‬ߥߖࠆੱ᧚߇㊀ⷐߢߔ‫ޕ‬

⽷࿅ᴺੱࡃࠗࠝࠗࡦ࠳ࠬ࠻࡝࡯දળ
ዊᨋ⦟ೣ
1947 ᐕ↢߹ࠇߩ࿅႙਎ઍߢߔ‫ޕ‬2008 ᐕ 6 ᦬߹ߢ࠳ࠗ
࠮࡞ൻቇᎿᬺ㧔ᩣ㧕ߢක⮎㑐ㅪ੐ᬺߩ⎇ⓥ࠹࡯ࡑડ↹߆ࠄၮ␆⎇ⓥ‫ޔ‬Ꮏᬺൻ⎇ⓥ߹ߢࠍᜂᒰߒߡ߅ࠅ߹
ߒߚ‫ޕ‬หᐕ 10 ᦬એ᧪‫ޔ‬NEDO ࡊࡠࠫࠚࠢ࠻‫ޟ‬㧔వዉᛛⴚ㐿⊒㧕㧛㉂⚛♧ൻ࡮⊒㉂ߦ⾗ߔࠆၮ⋚⎇ⓥ‫ߩޠ‬⎇ⓥߦᓥ੐ߒߡ᧪߹ߒߚ‫ߥ߁ࠃߩߎޕ‬ቇળߩ⊝ߐࠎᣇߦߪ޽߹ࠅ⋧ᔕߒߊߥ޿⹤㗴߆߽ߒࠇ߹ߖࠎ߇⎇ⓥ㐿⊒ߦ߅ߌࠆ‫⹏ޟ‬ଔ‫ߡ޿ߟߦޠ‬ዋߒㅀߴߡߺߚ޿
ߣᕁ޿߹ߔ‫ޕ‬ ᚒ‫⋡ߩ࠻ࠢࠚࠫࡠࡊߩޘ‬ᮡߪᚒ߇࿖⁛⥄ߩ㜞ᯏ⢻ࡃࠗࠝࡑࠬ♧ൻ㉂⚛㧔࠮࡞࡜࡯࠯㧕ߩഃᚑ‫ޔ‬㜞ല₸♧ൻᴺ෸߮㜞ല₸⊒㉂ᴺߩ㐿⊒ࠍ⋡ᜰߘ߁ߣߔࠆ߽
ߩߢߔ‫ᦨޕ‬ೋߦขࠅ⚵ࠎߛ࠹࡯ࡑߪ‫ޔ‬ฦ⒳࠮࡞࡜࡯
࠯ߩᵴᕈ⹏ଔᴺߩᮡḰൻߢߒߚ‫ߪࠇߎޕ‬ᢥ₂਄ߩ࠺࡯࠲ߛߌߢߪᵴᕈ᷹ቯ㧔⹏ଔ㧕ᣇᴺ߇߫ࠄ߫ࠄߢ‫ޔ‬ᧄᒰߦߤߩ㉂⚛߇ఝ⑲߆⺒ߺขࠇߥ޿ߎߣߦ㐓ߺ‫ޔ‬ฦ⒳㉂⚛ᵴᕈ᷹ቯߦߟ޿ߡᮡḰൻᴺࠍቯ߼‫ࠄࠇߎޔ‬ߦࠃࠅฦ⒳Ꮢ⽼㉂⚛߿⥄ࠄߩ㐿⊒㉂⚛ߩ⢻ജࠍหߓ‛Ꮕߒߢ⹏ଔߒࠃ߁ߣߔࠆ߽ߩߢߔ‫ߩߎޕ‬ਛߢ FPU
ᵴᕈ‫ޔ‬CMC ಽ⸃ᵴᕈ߿ࡃࠗࠝࡑࠬ♧ൻ⢻╬ߩᵴᕈ᷹
ቯᴺࠍᮡḰൻߒ߹ߒߚ‫⎇ߩߎޔߚ߹ޕ‬ⓥࠍㅴ߼ࠆ਄
ߢ‫ޔ‬ၮ⾰ߩᮡḰൻ߇㊀ⷐߣ⠨߃‫ ♽ᧄ⨲ޔ‬2 ⒳㧔Ⓑࠊࠄ‫ࠬࠨࡦࠕ࡝ࠛޔ‬㧕‫ ♽ᧄᧁޔ‬2 ⒳㧔࡙࡯ࠞ࡝‫᧖ޔ‬㧕ࠍขࠅ਄ߍ‫⧦ޔ‬ᕈ࠰࡯࠳࡮Ꮧ⎫㉄࡮᳓ᾲಣℂᴺߦࠃࠅ૞ᚑߒߚઍ⴫⊛․ᕈߩ೨ಣℂ‛ࠍᮡḰ೨ಣℂᮡຠߣߒߡቯ߼߹ߒߚ‫ޔߡߒߘޕ‬ᮡḰᵴᕈ᷹ቯᴺߣᮡḰ೨ಣℂᮡຠࠍ↪޿‫ޔ‬ฦ⒳Ꮢ⽼࠮࡞࡜࡯࠯ߣᚒ‫߇ޘ‬ഃᚑߒߚᡷ⦟㉂⚛ߩᕈ⢻ࠍᲧセ⹏ଔߒ߹ߒߚ‫⚿ߩߎޕ‬ᨐ‫ޔ‬ᦨ⚳⊛ߦᶏᄖ㉂⚛ࡔ࡯ࠞ࡯⵾ຠߩ♧ൻᕈ⢻ࠍಒ㚧ߔࠆ㜞ᯏ⢻㉂⚛ߩഃᚑߦᚑഞߒ߹ߒߚ‫⥄ޕ‬ಽߩ┙ߜ૏⟎ࠍ಄㕒ߦ‫ޔ‬ᱜ⏕ߦ‫౏ޔ‬ᐔߦ⹏ଔߢ߈ࠆ߆ߤ߁߆ߪ
࠹ࠢࡁࡠࠫ࡯ߦ߅޿ߡᚑഞߢ߈ࠆ߆ุ߆ߩࠠ࡯ࡐࠗ
ࡦ࠻ߢߔ‫ޕ‬46

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)40 ઍ߇૗ੱ߆޿߹ߔ‫ޔߚ߹ޕ‬ᢙੱߩ⧯⠪㆐߇ࡃࠗࠝ
ಽ㊁ߢߩᣂ੐ᬺ⏕┙ߦะߌߡ㗎ᒛߞߡ޿ࠆᆫ߽⷗ߡ
޿߹ߔ‫ࠅࠊ⚳ࠄߚߞߥߊߥ޿߇ੱࠆߔࠫࡦ࡟ࡖ࠴ޕ‬ߢߔ‫ੱߥ߁ࠃߩߎޕ‬㆐ߦߤࠎߥᚻഥߌ߇ߢ߈ࠆ߆ࠍ
⠨߃ታⴕߔࠆ੐߽ᚒ‫ޘ‬਎ઍߩ㊀ⷐߥ⺖㗴ߛߣᕁ޿߹
ߔ‫ᦨޕ‬ᓟߪ⣕✢ߒ߹ߒߚ‫ޕ‬એ਄ ᚒࠄ࿅႙ߩ਎ઍ߇⎸ߥ⛔ᴦ⢻ജࠍᜬߜวࠊߖߡ޿
ߥ޿ߎߣࠍ㔺๒ߒߚߎߎᢙᐕߢߒߚ‫ޕ‬ᱷ㑆㉿ᳯሶߐࠎ߇‫ޔ‬࿅႙ߩ਎ઍࠃ‫ࡃ࠰ޔ‬ᛂߜߦ⥝ߓߡ޿ࠆ႐ว߆
ߣ᜸⊒ߒߡ޿߹ߔ‫ޕ‬ᚒ‫ߩޘ‬਎ઍߪዋߥߊߣ߽ 70 ᱦ૏߹ߢߪ⧯⠪㆐ߩ㇎㝷ߦߥࠄߥ޿ࠃ߁ߦචಽ⇐ᗧߒߟߟ‫ޔ‬ᣣᧄߩ↥ᬺࠍ┙ߡ⋥ߔὑߦ⥄ಽ㆐ߦ૗߇ߢ߈ࠆ
߆ࠍ⠨߃‫ⴕޔ‬േߔߴ߈ߣ⠨߃߹ߔ‫ߩ⑳ޕ‬෹ੱߦਛዊ
ડᬺ߿ࡌࡦ࠴ࡖ࡯ߢ੐ᬺ⏕┙ߩὑߦᅗ㑵ߒߡ޿ࠆ⎇ⓥᦨ೨✢

⁛┙ⴕ᡽ᴺੱㄘᬺ↢‛⾗Ḯ⎇ⓥᚲ ੇ῎⠴ᕈ⎇ⓥࠣ࡞࡯ࡊ 㤛Ꮉ↰㓉ᵗࠆߎߣߦߥࠅ߹ߒߚ‫ޕ‬ਥⷐࡔࡦࡃ࡯ߣߒߡᅏ↰ඳ჻‫ޔ‬ᷰㆺආᒾ᳁‫ޔ߇⑳ߡߒߘޔ‬ฦኾ㐷ಽ㊁ߦಽ߆ࠇߡ⎇ⓥࠍࠬ࠲࡯࠻ߐߖ߹ߒߚ‫ޕ‬ ᦨೋߦ⌕ᚻߒߚ⎇ⓥߪ‫⫾ߦ⊛⇣․ᤨ῎ੇޔ‬Ⓧߔࠆਥⷐઍ⻢↥‛ߩหቯߢߒߚ‫ޕ‬ᷰㆺ᳁ߣ⑳ߪදജߒߡ‫ޔ‬♧‫ޔ‬⢽⾰‫⫾ߩ㉄ࡁࡒࠕޔ‬Ⓧᄌേࠍ⺞ߴߡ޿߈߹ߒߚ‫ޕ‬ߘߩ⚿ᨐ‫ޔ‬㧔ඨ੍߫ᗐㅢࠅߢߒߚ߇㧕↢૕ᚑಽ଻⼔ᯏ⢻ࠍᜬߟߎߣߢ⍮ࠄࠇߡ޿ࠆ㕖ㆶరᕈੑ♧ߩ࠻࡟ࡂࡠ࡯ࠬ߇ᄢ㊂ߦ⫾Ⓧߒߡ޿ࠆ੐ࠍ⷗޿ߛߒ߹ߒߚ‫ޕ‬ᷰㆺ᳁ߪ᣸⯻↢ℂቇߩࠛࠠࠬࡄ࡯࠻ߢ޽ߞߚߚ߼‫ޔ‬૕㐳㧣mm ᒙߩዊߐߥᐜ⯻߆ࠄ⥄࿷ߦ⚵❱ࠍขࠅ಴ ‫ߦ࡜ࠞ࡜ࠞޟ‬ᐓ߆ࠄ߮ߡ߽ᱫߥߥ޿↢‛߇޿ࠆ‫ޕ‬‫ޠ‬ࡀࡓ࡝࡙ࠬ࡝ࠞߩ⛮ઍ㘺⢒ᴺࠍ⏕┙ߐߖߚ߫߆ࠅߩ
ᅏ↰㓉ඳ჻߆ࠄߎߩ⹤ࠍᆎ߼ߡ⡞޿ߚߩߪ‫ޔ‬2000 ᐕ
ߩ⑺ߢߒߚ‫ߩߛߚޕ‬⍹☸ߩࠃ߁ߦߺ߃ࠆੇ῎ߒߚᐜ⯻߇‫ޔ‬᳓ߦᶐߒߚߛߌߢ⷗ࠆ⷗ࠆ߁ߜߦๆ᳓ߒᕷࠍ
็߈㄰ߒߡ޿ߊ᭽ߦ‫߁޿ߣߞ޽ޔ‬㑆ߦ㝯ੌߐࠇߚߩ
ߪ޿߁߹ߢ߽޽ࠅ߹ߖࠎ‫ߩߘޕ‬⍍㑆‫⎇ޔ‬ⓥੱ↢ߩోߡ⾲ߌߡ߽୯ߔࠆ⎇ⓥኻ⽎ߢ޽ࠆߎߣࠍታᗵߒ߹ߒ
ߚ‫⎇ߩߢ߹ࠇߘߚ޿↪ࠍࠦࠗࠞޕ‬ⓥࠍࠬ࠶ࡄ࡝ߣ߿߼‫῎ੇߩࠞ࡝࡙ࠬ࡝ࡓࡀޔ‬⠴ᕈᯏ᭴ߩ⎇ⓥߦᴚ㗡ߔ

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)޿߹ߔ‫⃻ޕ‬࿷ߢ߽ෆᲣ⚦⢩⊒⃻♽ߪ‫ޔ‬ᚒ‫ߢࡏ࡜ߩޘ‬ߩㆮવሶᯏ⢻⸃ᨆߩၮᐙᛛⴚߣߒߡ⛮ᛚߐࠇߡ޿߹ߔ‫ޕ‬ ᤨߪ⚻ߜ‫⃻ޔ‬࿷ߢߪ⎇ⓥቶߩࡔࡦࡃ࡯߽Ⴧ߃‫ޔ‬2007ᐕߦࡀࡓ࡝࡙ࠬ࡝ࠞߩㅒㆮવቇ⊛⸃ᨆ㧔RNAi ߿ㆮવሶዉ౉࡙ࠬ࡝ࠞߩ᭴▽㧕ࠍⴕߞߡ޿ࠆࡈ࡜ࡦࠬੱ
⎇ⓥຬߩ Richard Cornette ඳ჻߇ടࠊࠅ‫ޔ‬2011 ᐕߦߪ⚵❱ൻቇߩࠬࡍࠪࡖ࡝ࠬ࠻ߢ޽ࠆᔒ᧛ᐘሶඳ჻߇
ടࠊࠅ߹ߒߚ‫⎇ࠢ࠼ࠬࡐޔߚ߹ޕ‬ⓥຬ߽᭽‫ߥޘ‬ᣇ‫ޘ‬߇࿷☋ߒߡ߈߹ߒߚ߇‫⃻ޔ‬࿷ߪࡀࡓ࡝࡙ࠬ࡝ࠞߩࠥ
ࡁࡓ߿ࡔ࠲ࡏࡠࡒࠢࠬ⸃ᨆࠍⴕߞߡ޿ࠆࡠࠪࠕੱ⎇ⓥຬߩ Oleg Gusev ඳ჻‫ޔ‬ᣂⷙ࠻࡟ࡂࡠ࡯ࠬ࠻࡜ࡦࠬ
ࡐ࡯࠲࡯ߩหቯߣ࠻࡟ࡂࡠ࡯ࠬߩ in vivo ࠗࡔ࡯ࠫࡦࠣࠍ⎇ⓥߒߡ޿ࠆ JSPS-PD ߩ⩵↰⌀๋ඳ჻‫ߒߘޔ‬ߡ LEA ࠲ࡦࡄࠢ⾰ߩᯏ⢻⸃ᨆࠍㅴ߼ߡ޿ࠆ JSPS-PDߩ⇌ਛℂᕺඳ჻ߣ৻✜ߦ⎇ⓥࠍㅴ߼ߡ޿߹ߔ‫ޕ‬ ࿖㓙⦡⼾߆ߥࡔࡦࡃ࡯ߣ౒ߦ‫ߣࠞ࡝࡙ࠬ࡝ࡓࡀޔ‬޿߁ᄸᅱߥ᣸⯻ߩోߡࠍ⍮ࠆߚ߼ߦ‫⸃ࠬࠢࡒ࡯ࠝޔ‬ᨆ߆ࠄㆮવሶᯏ⢻⸃ᨆ‫ޔ‬ၭ㙃⚦⢩ߩ᮸┙‫ޔ‬శቇ㗼ᓸ
㏜߿㔚ሶ㗼ᓸ㏜ࠍ↪޿ߚ⚦⢩࡮⚵❱ൻቇ⊛⸃ᨆ‫ޔ‬ᨐߡߪࡀࡓ࡝࡙ࠬ࡝ࠞߩੇ῎⠴ᕈᯏ⢻ࠍᔕ↪ߒߚ↢૕ᚑಽߩᏱ᷷ੇ῎଻ሽᴺߩ᭴▽߽ⴕߞߡ޿߹ߔ‫ࡓࡀޕ‬࡝࡙ࠬ࡝ࠞ⎇ⓥߪ‫ޔ‬਎⇇ࠍ⷗࿁ߒߡ߽ᚒ‫⎇ߩޘ‬ⓥቶ
ߢߒ߆ⴕߞߡ޿߹ߖࠎ‫ߩ࡯ࠖ࠹࡝࠽ࠫ࡝ࠝޕ‬ઍఘߣߒߡ‫⎇ߩߡోޔ‬ⓥ㐿⊒ࠍ⥄ࠄ߇ⴕࠊߥߌࠇ߫޿ߌ߹
ߖࠎ‫ޕ‬㗬ࠆ߽ߩ߇ઁߦ޿ߥ޿ߎߣߪᄢᄌߢߔ߇‫ߛޔ‬߆ࠄߎߘ‫⎇߽ߡߌ⾲߽ߡోߪ⑳ޔ‬ⓥߔࠆߦ୯ߔࠆߣ
ᕁߞߡ޿߹ߔ‫⁛ޕ‬ᴺߩ⎇ⓥᚲߣ⸒߁ߎߣ߽޽ࠅ‫ޔ‬ቇ↢ߐࠎ߇ᚲዻߔࠆߎߣߪṌᄙߦ޽ࠅ߹ߖࠎ߇‫ޔ‬ㄘᬺ↢‛⾗Ḯ⎇ⓥᚲߪ᧲ᄢᄢቇ㒮ߩㅪ៤⻠ᐳࠍᜬߞߡ޿
߹ߔߩߢ‫⎇ߩࠞ࡝࡙ࠬ࡝ࡓࡀޔ‬ⓥߦ⥝๧ࠍᜬߚࠇߚ
ቇ↢ળຬߩᣇ‫ޔ߇ޘ‬ᚒ‫⎇ߩޘ‬ⓥߦෳടߐߖࠆߎߣࠍ
ᒝߊᦸࠎߢ޿߹ߔ‫ޔߚ߹ޕ‬ታ‛ߩࡀࡓ࡝࡙ࠬ࡝ࠞࠍ
ߺߚ޿ߣᏗᦸߐࠇࠇ߫޿ߟߢ߽᱑ㄫߒ߹ߔߩߢ‫⎇ޔ‬ⓥቶࠍ⷗ቇߦ᧪ߡߛߐ޿‫ޕ‬ߔߎߣ߇ߢ߈߹ߒߚ‫ߩߘޕ‬ථ⿧ߒߚᛛⴚࠍ↪޿‫ޔ‬ᢿ㗡ߒߚᐜ⯻ߢ޽ߞߡ߽‫៰ޔ‬಴ߒߚ⢽⢌૕㧔ູ੃േ‛ߩ⢄⤳ߦ⋧ᒰߔࠆེቭ㧕ߢ޽ߞߡ߽‫῎ੇޔ‬ೝỗߢ࠻࡟ࡂࡠ࡯ࠬࠍ⫾Ⓧߒߡήઍ⻢ભ⌁㧔ࠕࡦࡅ࠼ࡠࡆࠝ
ࠪࠬ‫߽ߣ⌁ੇޔ‬๭߫ࠇࠆ㧕ߩ⁁ᘒߦߥࠆߎߣࠍ⊒⷗ߒ߹ߒߚ‫⚻␹ޔߪࠞ࡝࡙ࠬ࡝ࡓࡀޔࠄ߆ߣߎߩߎޕ‬♽߿ౝಽᴲ♽ߩ㑐ਈήߒߦ‫ޔ‬ฦ⚵❱࡮⚦⢩߇⥄⊒⊛ߦੇ῎ήઍ⻢ભ⌁ࠍ⺃ዉน⢻ߢ޽ࠆߎߣ߇␜ໂߐࠇ
߹ߒߚ‫ޕ‬
 ߘߩᓟ‫ޔ‬ᅏ↰ඳ჻ߣᷰㆺ᳁ߪ‫ߩࠞ࡝࡙ࠬ࡝ࡓࡀޔ‬᡼኿✢⠴ᕈߩ↢ℂቇ⊛⎇ⓥࠍㅴዷߐߖ‫῎ੇޔ‬ᐜ⯻ߪ
10k Gy ߽ߩ㜞޿᡼኿✢㧔ǫ✢߿ࡋ࡝࠙ࡓࠗࠝࡦࡆ࡯
ࡓ㧕ࠍᶎ߮ߡ߽‫ޔ‬ๆ᳓⯃↢ߒ‫ߩߘޔ‬ᓟߩ↢⢒߇น⢻ߢ޽ࠆߎߣࠍ␜ߒ߹ߒߚ‫৻ޕ‬ᣇ‫ޔߪ⑳ޔ‬ኾ㐷ಽ㊁ߢ޽ࠆಽሶ↢‛ቇ࡮↢ൻቇߩⷰὐ߆ࠄήઍ⻢ભ⌁ߩࡔࠞ࠾࠭ࡓࠍℂ⸃ߔࠆߎߣߦദ߼߹ߒߚ‫ޕ‬ᰴ਎ઍࠪ࡯
ࠤࡦࠨ࡯߇㓉⋓ࠍ⹶ࠆ⃻࿷ߢߪ⠨߃ࠄࠇ߹ߖࠎ߇‫ޔ‬ᄙ㗵ߩ⾌↪ߣ 3 ᐕ߽ߩᤨ㑆ࠍ߆ߌߡࠨࡦࠟ࡯ဳ
DNA ࠪ࡯ࠤࡦࠨ࡯ࠍ↪޿ߚ EST ࠺࡯࠲ࡌ࡯ࠬߩ᭴
▽ࠍߒ߹ߒߚ‫ߩߘޕ‬ㆊ⒟ߢ‫ޔ‬ᄙ⚦⢩↢‛߆ࠄೋ߼ߡ
࠻࡟ࡂࡠ࡯ࠬ࠻࡜ࡦࠬࡐ࡯࠲࡯TRET1 ࠍ⊒⷗ߒ‫ࠕޔ‬ࠢࠕࡐ࡝ࡦ㧔᳓࠴ࡖࡀ࡞㧕߇ࡀࡓ࡝࡙ࠬ࡝ࠞ૕ౝ߆
ࠄߩ᳓ಽ⫳⊒ࠍଦㅴߐߖࠆߎߣࠍ⷗޿ߛߒ‫ޔ‬ᬀ‛ߦߒ߆ሽ࿷ߒߥ޿ߣᕁࠊࠇߡ޿ߚੇ῎⠴ᕈ㑐ㅪ࿃ሶߢ޽ࠆ LEA ࠲ࡦࡄࠢ⾰ࠍ᣸⯻߆ࠄೋ߼ߡหቯߒ߹ߒ
ߚ‫ޕ‬ㆮવሶࠍหቯߒߡ߽‫ޔ‬ᯏ⢻ࠍ⏕⹺ߒߥߌࠇ߫ᧄᒰߦੇ῎ήઍ⻢ભ⌁ߦ㊀ⷐߥߩ߆ߤ߁߆ಽ߆ࠅ߹ߖ
ࠎ‫ࠢࡄࡦ࠲⤑ߩߤߥ࡞ࡀࡖ࠴߿࡯࠲࡯ࡐࠬࡦ࡜࠻ޕ‬⾰ߩᯏ⢻⸃ᨆߩ႐㕙ߢ‫࡞ࠛࠟࡔ࠷ࠞ࡝ࡈࠕޔ‬ෆᲣ⚦⢩⊒⃻♽ߪᒝജߥ࠷࡯࡞ߦߥࠅ߹ߔ‫ߩߎޕ‬ታ㛎ࠍᒰᤨጤᚻᄢቇᄢቇ㒮↢ߛߞߚᢧ⮮ᓀሶߐࠎ߇ⴕߞߡߊ
ࠇ߹ߒߚ‫ࠬ࡯ࡠࡂ࡟࠻ޕ‬ṁᶧߦᶐߒߚ TRET1 ⊒⃻ෆᲣ⚦⢩ߩ☻᛽಴ᶧࠍ↪޿ߡ♧ಽᨆࠍⴕߞߚߣ߈‫ޔ‬
HPLC ߩࠢࡠࡑ࠻࠴ࡖ࡯࠻ߦ࠻࡟ࡂࡠ࡯ࠬߩࡇ࡯ࠢ
ࠍ⏕⹺ߒ‫⁅ޔ‬༑ੂ⥰ߒߚߎߣࠍ੹ߢ߽㞲᣿ߦⷡ߃ߡ

48

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪦㪩㪠㪞㪠㪥㪘㪣㩷㪧㪘㪧㪜㪩㩷

Journal of Japanese Society for Extremophiles (2012) Vol.11, 49-58

Zhang Xa, Nakahara Ta, Ochiai Nb, Kawauchi Ac, Hashimoto Jc,
Inoue Tc and Kudo Tc㩷

Diversity and function of aerobic culturable bacteria
in the intestine of deep-sea holothurianaGraduate School of Science and Technology, Nagasaki University, 1-14 Bunkyo-cho, Nagasaki 852-8521, Japan
Faculties of Fisheries, Nagasaki University, 1-14 Bunkyo-cho, Nagasaki 852-8521, Japanc
Graduate School of Fisheries Science and Environmental Studies, Nagasaki University, 1-14 Bunkyo-cho, Nagasaki
852-8521, Japanb

Corresponding author: Toshiaki KUDO, kudot@nagasaki-u.ac.jp
Phone & Fax: +81-95-819-2838
Received: August 17, 2012 / Accepted: October 4, 2012
bacteria6,7,11).
Gut bacteria play an important role in digestion of diets.
Studies on bacteria associated with Holothurians werereported only for Holothuria atra10) and Molpadia
musculus1,2). Ward-Rainey et al. reported partial aerobic
bacterial flora of Holothuria atra. 10) In their report, only
23 isolates were characterized by 16S rRNA genesequences analysis (the first 300 nucleotides) and they
were affiliated to the genera Vibrio and Bacillus. On theother hands, Amaro et al. used non-culturing methods toanalyze bacterial community of abyssal holothurian,Molpadia musculus.2) Their results suggested that the gut
bacterial composition was similar to that of the organic
matter-rich sediments. Members of CytophagaFlavobacteria-Bacteroides (CFB) group dominated in thebacterial community2). Recently, they also found that ca.
82% of total bacterial OTUs (Operational TaxonomicUnit) were common between the gut contents and thesurrounding sediments1). Enomoto et al. also reported
recently that J-Proteobacteria members were mainly
isolated as culturable bacteria from the intestine of
Apostichopus japonicus3). Using the moleculartechniques, they also found that Proteobacteria members
were main metabolically active microbial populations in
the intestine of Apostichopus japonicus.
Gut microorganisms play an important role in digestionof diets, but the diversity and function of aerobic
culturable bacteria in the intestine of the deep-sea
holothurian are still unclear.
In this report, we isolated ninety-two aerobic culturable
bacterial strains from each part of the intestine of the
deep-sea holothurian collected at the southeast of Fukue

Abstract
Ninety-two aerobic culturable bacterial
strains were isolated from each part of the intestine of the
deep-sea holothurian collected at 32°30N, 129°09E(southeast of Fukue Island, Nagasaki, Japan) and water
depth of 236 m in November 2010. The temperature of
seawater at the water depth of the sampling point wasestimated to be ca.13-14 °C from data in Japan
Meteorological Agency. By partial 16S rRNA genesequences of the isolates, the isolates belonged to 45
nearest type strain species (below, referred to as species).
High diversity was observed in the genera Bacillus (21species) and Vibrio (6 species). The bacterial diversity
was similar among three parts, i.e. anterior, mid andposterior parts of the intestine and 14 species weredetected in multiple parts of the intestine. Most isolates
showed various polysaccharide degradation activities but
few isolates showed alginate or agar degradation
activities probably because there were no seaweeds
containing alginate or agar in this deep-sea. Comparing
the functions and properties of several species in threeparts, the posterior part was likely to be different fromthe anterior or mid parts.
Key words: aerobic culturable bacteria; deep-sea
holothurian; diversity; function; 16S rRNA gene
Introduction
Deep-sea is one of unexplored regions even today.
Holothurians (~1430 species) are found on various sea
floors from deep sea floors to intertidal areas 4,9).
Holothurians belong to phylum Echinodermata and theirdiet is detritus such as organic matter, microalgae, and49

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)southeast of Fukue Island, Nagasaki, Japan (32°30N,
129°09E), at a water depth of 236 m in November 21,
2010 (Fig.1). The temperature of seawater at the water
depth of the sampling point was estimated to be
ca.13-14 °C from data in Japan Meteorological Agency.
The specimen was kept in icebox and asepticallydissected in our laboratory in November 24, 2010.
Whole intestine was excised from the animal body
aseptically using sterilized instruments. Fraction ofintestine was carried out according to Shimizu et al. 8)
The intact intestine was divided into three parts, theanterior part (0.71g), the mid part (0.88g) and theposterior part (1.80g) (Fig. 2). To isolate bacteria fromboth intestinal wall and contents, 1 ml of saline wasadded to each part and each part was crushed and mixed
enough. Each suspension thus obtained was used for
isolation of bacteria and 50 㱘l of the each suspension
was spread on plates.

Island, Nagasaki, Japan, water depth of 236 m and in
November 2010. We found that the aerobic culturable
isolates belonged to 45 nearest type strain species (below,
referred to as species). The bacterial diversity was
similar among three parts, i.e. anterior, mid and posterior
parts of the intestine. Most isolates showed various
polysaccharide degradation activities but few isolates
showed alginate or agar degradation activities probably
because there were no seaweeds in deep-sea. On theother hand, when we compared the functions andproperties of several species in three parts, the posterior
part was likely to be different from the anterior or mid
parts. Maybe the posterior part was related to thedigestion of polysaccharides or high salt environment.
Materials and Methods
Sample collection and dissection
Deep-sea holothurian specimen was collected at

Fig.1 Map of sampling site (arrow) and environmental conditions.
Sampling point : 32°30N, 129°09E (Southeast of Fukue Island, Nagasaki, Japan) Water depth: 236 m, Sampling data:
November 21, 2010, Temperature of water depth 200 m : ca.13-14㷄, Sea surface temperature: ca. 21-22㷄䋨Data fromJapan Meteorological Agency䋩tesque, Kyoto, Japan) were added to Horikoshi medium
as carbon sources (final concentration 1%).
LB solid medium (pH 7) contained 1% tryptone (Difco),
0.5% yeast extract (Difco), 3.5% NaCl, and 1.5 % agar
(Wako pure chemicals, Osaka, Japan). Horikoshi solid
medium (pH 7) contained 1% polysaccharide, 0.5%
peptone (BD), 0.5% yeast extract, 0.1% KH2PO4, 0.02%

Growth media
Luria-Bertani medium (LB) and Horikoshi medium
were used basically. But NaCl concentration was 3.5%
instead of 1%. Polysaccharides such as carboxymethyl
cellulose sodium salt (CMC) (Wako pure chemicals,
Osaka, Japan), xylan (Sigma), sodium alginate (Wakopure chemicals, Osaka, Japan) and soluble starch (nacalai

50

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)1-2. Cellulase detection: Basic neutral agar medium forcellulase detection contained 0.1% CMC, 3.7% marine
broth, 0.6% MgCl2 䊶 6H2O, 1.5% agar, 1.6% NaCl,
0.0015% congo-red, adjust pH to 7.0 with 1N NaOH.
Clear zone around a colony suggested cellulase activity.
1-3. Alginate lyase detection: The basic neutral agar
medium for alginate lyase detection, contained 1%
sodium alginate, 3% NaCl, 0.07% KCl, 0.26% MgSO4,
0.5% MgCl2, 0.1% CaSO4, 0.5% peptone, 0.01% ferricphosphate, 0.1% yeast extract, 2% agar; adjust the pH to
7.0 with 1N NaOH. After two weeks’ incubation at 30 °C,
70% ethanol was filled into plates. A clear zone aroundthe colony indicated the presence of alginate lyase.
1-4. Xylanase detection: Horikoshi agar medium
containing 1% xylan was used for xylanase detection.
Xylanase-producing colony showed clear zone around acolony.
1-5. Agarase detection: Horikoshi agar medium without
polysaccharide was used for agarase detection.
Agarase-producing colony showed dent around a colony.
2. Alkaline agar plates
For alkaline agar plates, Na2CO3 (autoclaved separately)
was added to neutral agar medium (final pH:
pH10.3-10.5). Na2CO3 concentration of alkaline plate
was 1%.

MgSO4•7H2O, 3.5% NaCl, and 2% agar. Sodium alginate
solid medium contained 2.5% agar. For 10% NaCl media,
NaCl concentration of growth media was 10% instead of
3.5%.
For alkaline agar plates, Na2CO3(autoclaved
separately) was added to neutral agar medium (final pH:
pH10.3-10.5). Na2CO3 concentration of alkaline plate
was 1%.
Isolation of bacteria
Fifty l of the gut suspensions was directly plated onagar plates without enrichment culture. High salt
concentration or high pH were used for isolation
conditions to isolate various bacteria because marine
water is semi-alkaline pH and contains 3.5% NaCl.
Seventeen different media were prepared by combinationof pH, NaCl concentration and carbon source (Table 1).
The plates were incubated at 30 °C aerobically for two
weeks to obtain slowly growing bacteria. Bacteria wereisolated from each plate, purified and stored in slants forfurther analysis.

All isolates were tested for salt tolerance: 0%, 3.5%,
10%, 15%, 20%, 25% NaCl (w/v), pH tolerance (pH7and pH10) and effect of oxygen.
Anaerobic growth was examined using gaspak(COSMO BIO) at 30 °C for two weeks, and then growth
condition was changed to the aerobic condition at 30°C
for two weeks. The isolates were assigned to threegroups, facultative anaerobic bacteria (FA), anaerobic
tolerant bacteria (AT) and aerobic bacteria (A).
Facultative anaerobic bacteria form colony in both
aerobic and anaerobic cultivation. Anaerobic tolerant
bacteria do not form colony in anaerobic condition for
two weeks but form colony in aerobic cultivation after
the anaerobic cultivation. Aerobic bacteria do not formcolony in anaerobic condition for two weeks and also do
not form colony in aerobic cultivation after the anaerobic
cultivation. Growth ability at various conditions ofsalinity or pH was measured at 30 °C for two weeks. The
isolates were divided into two groups by effect of pH on
growth, neutrophilic bacteria (NE) that grew only at pH7,
and alkaliphilic/alkali-tolerant bacteria (ALK) that grewboth at pH 7 and pH10.

Physiological and biochemical characteristics ofisolates
Polysaccharide degradation activities were detected byplate methods using CMC, xylan, alginate, starch or agar
as substrate. The following plates were prepared for
detection of enzyme activities.
1.
Neutral agar plates
1-1. Amylase detection: Horikoshi agar medium
containing 1% potato starch instead of soluble starch wasused for amylase detection. Amylase-producing colony
showed turbid halo around a colony.

51

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)obtained in Horikoshi media (pH7), but lower cfu
numbers in the alkaline Horikoshi media (ca. pH10.3).
Twenty-four, 33 and 35 isolates were obtained from theanterior, mid and posterior suspensions, respectively. In
total, 92 isolates were purified and analyzed further.

Molecular identification of the isolates
Partial analysis of 16S ribosomal RNA (rRNA) gene of
the isolates was carried out. The 16S rRNA gene wasamplifiedusingbacterial
primers
27f(5’-AGAGTTTGATCCTGGCTCAG-3’) and 1492r(5’-GGTTACCTT GTTACGACTT-3’) and the purified
PCR product was sequenced with dideoxynucleotide
chain-termination method using 3130 or 3730 DNA
sequencer (Applied Biosystems). Primers 27F, 520R
(5’-ACCGCGGCT
GCTGGC-3’)
and907R(5’-CCGTCAATTCMTTTRAGTTT-3’) were used ingene sequencing reactions. Sequences of the partial 16S
rRNA genes were assembled and edited using
Sequencher (version 4.10.1 demo, Gene Codes
Corporation) and MacVector (version 10.0.2).
Nucleotide sequences of the partial 16S rRNA genes
have been submitted to GenBank/EMBL/DDBJ
databases under accession numbers AB741781 throughAB741873 except AB741857 (Supplementary Table 1).
The partial 16S rRNA gene sequences were comparedwith other sequences in DDBJ database using BLAST
program and compared with type strain sequences in
Ribosomal database project (RDP). Each isolate was
assigned to nearest type strain species (Supplementary
Table 1). When isolate showed more than 97% identities
with some type strain sequences, the isolate was assigned
to nearest type strain species (below, referred to as
species). When isolate showed less than 97% identities
with any type strain sequences, the isolate was assigned
to the tentative nearest type strain species (below,
referred to as tentative species).

Fig. 2 Photographs of the deep-sea holothurian (dorsalside A, ventral side B) and its dissection (C), including
anterior intestine (1), mid intestine (2), posterior intestine(3), Polian vesicle (4) and respiratory trees (5). Cuvierian
tubules were not detected in the sample.

Results
Phylogenic analysis of bacterial isolates
The partial 16S rRNA gene sequences were done andcompared with other sequences in DDBJ database using
BLAST program and compared with the type strain
sequences in Ribosomal database project (RDP). Table 2
summarized the species/the tentative species of theisolates as determined via BLAST (Supplementary Table1). By partial 16S rRNA gene sequences of the isolates,
the isolates belonged to 45 species. Fourteen species
were detected in multiple locations of three parts, i.e.
anterior, mid and posterior parts of the intestine. The
isolates belonged to the phyla Firmicutes (33 species)
and Proteobacteria (12 species) (Table 2). Among 33
species of the phylum Firmicutes, 21 species belonged to
the family Bacillaceae 1, the genus Bacillus. Ten species
belonged to the family Bacillaceae 2, the genera
Gracilibacillus,
Halobacillus,
Oceanobacillus,
Thalassobacillus and Virgibacillus. Twelve species of the

Isolation of bacteria
Fig.2 shows photographs of the deep-sea holothurian
and its dissection (C), including anterior intestine (1),
mid intestine (2), posterior intestine (3), Polian vesicle(4) and respiratory trees (5). But Cuvierian tubules were
not detected in the specimen. The each intestine
suspension was directly plated on agar plates andseventeen isolation media were used. Table 1
summarized number of the isolates obtained by different
cultural conditions. The intact intestine was divided into
three parts, the anterior part, the mid part and theposterior part. The each part was crushed and mixed
enough and the suspensions thus obtained were used for
isolation of bacteria. Number of colony forming units(cfu) per g of the anterior, mid and posterior parts were1.4㬍104 cfu/g, 0.6㬍104 cfu/g and 0.58㬍104 cfu/g in
LB medium, respectively. Similar cfu numbers were52

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)phylum Proteobacteria belonged to the genera Vibrio,
Halomonas, Photobacterium, Pseudomonas andMarinobacter. Among them, high diversity was found in
the genera Bacillus and Vibrio. The closest relatives ofthese isolates were observed in various sea environments.
The bacterial diversity was similar among three parts, i.e.
anterior, mid and posterior parts of the intestine (Fig.3)
and 14 species (indicated by star in Table 2) weredetected in multiple parts of the intestine. But, thenumber of species belonged to the family Bacillaceae 2
decreased in the posterior part of the intestine comparedwith those in the anterior or mid parts of the intestine(Fig.3).
Three isolates (isolate no. C214, C254 and C271)
showed less than 97% identities with any type strain
sequences. The isolates were assigned to the tentative
species. (Supplementary Table 1).

53

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Fig.3 Number of the species of isolates in each part of the intestine
The species were divided into five groups: the family Bacillaceae 1 (gray box), the family Bacillaceae 2 (vertical
stripes), the family Planococcaceae (dotted box), the family Staphylococcaceae (black box) and the order
J-Proteobacteria (diagonal stripes box). Fig.3 was summarized from Supplementary Tables 1.

such as Jeotgalibacillus campisalis, Vibrio pomeroyi andV. rotiferianus were found in multiple locations. Only 3
isolates producing alginate-lyase were affiliated with
Gracilibacillus dipsosauri, Pseudomonas synxantha andVibrio agarivorans. All isolates producing xylanase weremost closely related to the genera Bacillus (9 species)
and Pseudomonas (3 species). One species,
Pseudomonas libanensis was found in multiple locations.
Several species showed multiple polysaccharides
degradation activities. On the other hand, 11 species had
no polysaccharides degradation ability and 7 species ofthem in the anterior or mid parts of the intestinebelonged to the family Bacillaceae 2 but the number ofthem decreased in the posterior part of the intestine(Fig.4). It was observed that in the posterior part, thenumber of xylan degrading species related to the family
Bacilliaceae 1 increased and the number of starchdegrading species related to the order J-Proteobacteria
also increased (Fig.4).

Polysaccharide degradation ability of isolates
Many isolates from the specimen showedpolysaccharides degradation ability and degraded one ormore substrates (S, CMC, AL and XL). Twenty-eight, 12,
3 and 12 species showed amylase activity, cellulase
activity, alginate lyase activity and xylanase activity,
respectively (Fig.4, Supplementary Table 1). No agarase
activity was observed in all isolates.
Amylase producing isolates were mainly affiliated withthe genus Bacillus (15 species). Twelve species such as
Bacillus horikoshii, B. hunanensis, B. licheniformis, B.
megaterium,B.
vietnamensis,
Halobacillus
kuroshimensis, H. trueperi, Photobacterium rosenbergii,
Pseudomonas cedrina, P. libanensis, Vibrio pomeroyi,
and V. rotiferianus were found in multiple locations of
three parts, i.e. anterior, mid and posterior parts of theintestine.
Most cellulase positive isolates belonged to the genera
Bacillus (5 species) and Vibrio (4species). Three species

54

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Fig.4 Number of the species of isolates degrading various polysaccharides
The species were divided into five groups as shown in Fig.3. S, CMC, AL, XL and None indicated starch degradation
activity (S), CMC degradation activity (CMC), alginate degradation activity (AL), xylan degradation activity (XL) andno degradation activity (None), respectively. No isolates showed agarase activity. Fig.4 was summarized from
Supplementary Tables 1.
Thirty AT species were mainly affiliated to the genera
Bacillus (16 species), Gracilibacillus, Halobacillus,
Oceanobacillus and Virgibacillus. Only one aerobic (A)species, Halobacillus kuroshimensis was found in
multiple locations. The species belonging to the family
Bacillaceae 2 were mainly affiliated to AT group andspecies belonging to the family Bacillaceae 1 weremainly affiliated to FA or AT groups (Fig.5). The species
belonging to the phylum Proteobacteria were mainly
affiliated to FA group (Fig.5).

Physiological characteristics of the isolates
Fig.5 and Supplementary Table 1 showed effect ofanaerobic condition for growth of the isolates. The
isolates were divided into three groups, facultative
anaerobic bacteria (FA), anaerobic tolerant bacteria (AT)and aerobic bacteria (A). Twenty-five FA species weremainly affiliated with the phylum Proteobacteria and thefamily Bacillaceae 1. Six species such as Bacillus
licheniformis,
Photobacteriumrosenbergii,
Pseudomonas cedrina, P. libanensis, Vibrio pomeroyi
and V. rotiferianus were found in multiple locations.

Fig.5 Number of the species of isolates classified by effect of oxygen on the growth
The species were divided into five groups as shown in Fig.3. Facultative anaerobic bacteria (FA) form colony in both
aerobic and anaerobic cultivation. Anaerobic tolerant bacteria (AT) do not form colony in anaerobic condition for two
weeks but form colony in aerobic cultivation after the anaerobic cultivation. Aerobic bacteria (A) do not form colony inanaerobic condition for two weeks and also do not form colony in aerobic cultivation after the anaerobic cultivation.
Fig.5 was summarized from Supplementary Tables 1.
55

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Bacillaceae 1, the genus Bacillus and 9 species belonged
to J-proteobacteria such as the genera Pseudomonas,
Vibrio and Photobacterium.Seven species were slight
halophiles (3.5% NaCl) belonged to the genera Vibrio,
Photobacterium, Jeotgalibacillus Bacillus. It appears thatthe species belonging to the family Bacillaceae 2 aremost salt-tolerant (more than 20% NaCl) and thosebelonging to the phylum Proteobacteria or the family
Bacillaceae 1 were moderate halophilic (10-) (Fig.6). It
was observed that number of species related to slight
halophiles (3.5%) decreased in the posterior partcompared with those in other parts.

High salt concentration or high pH were used for
isolation conditions to isolate various bacteria because
marine water is semi-alkaline pH and contains 3.5%
NaCl. Salinity tolerance of the isolates was examined(Fig.6, and Supplementary Table 1; JJSE Web site). The
isolates most closely related to Halobacillus
kuroshimensis and Bacillus clausii showed the highest
salt tolerance (25 % NaCl) and some strains were able togrow in absent of NaCl. Twelve species were halophilic(20-25% NaCl conc.) and mainly belonged to the family
Bacillaceae 2, the genera Halobacillus, Virgibacillus andOceanobacillus. Thirty-two species were moderate
halophilic (10-) and 18 species belonged to the family

Fig.6 Number of the species of isolates classified by effect of NaCl concentration on the growth
The species were divided into five groups as shown in Fig.3. Fig.6 was summarized from Supplementary Tables 1.
45 species. The bacterial diversity was similar among
three parts, i.e. anterior, mid and posterior parts of theintestine and 14 species were detected in multiple partsof the intestine. But, the number of species belonged to
the family Bacillaceae 2 decreased in the posterior partof the intestine compared with those in the anterior or
mid parts of the intestine. (We will discuss this later.)
As shown in Table2, the isolates belonged to the phyla
Firmicutes (33 species) and Proteobacteria (12 species).
Among 33 species of the phylum Firmicutes, 21 species
belonged to the family Bacillaceae 1, the genus Bacillus.
Recently, Enomoto et al. reported that Proteobacteria
members were mainly isolated as culturable bacteria
from the intestine of Apostichopus japonicus3). These
results suggested that the sea environments such as deepsea or intertidal areas maybe affected diversity of aerobic
culturable bacteria.
Detritus is organic materials and is used as a source ofnutrient for detritus feeders5). Most important

All isolates were examined for growth responses to pH
shift(pH7pH10or
pH10pH7.
All
alkaliphilic/alkali-tolerant strains isolated from alkali
medium were able to grow at pH 7, while half of species
isolated from pH 7 were able to grow at pH 10. Nineteen
species were the neutrophilic species (NE) growing onlyat pH7. Eleven species were affiliated with the genus
Bacillus.
Discussion
In this report, we isolated aerobic culturable bacteria
from each part of the gut of deep-sea holothurian using
different culture conditions. The deep-sea holothurian
was collected at southeast of Fukue Island, Nagasaki,
Japan (32°30N, 129°09E), at a water depth of 236 m in
November 21, 2010. Ninety-two aerobic culturable
bacterial strains were isolated from each part of theintestine of deep-sea holothurian. By partial 16S rRNA
gene sequences of the isolates, the isolates belonged to56

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)In this study, only one specimen, deep-sea holothurian
was obtained and investigated. Therefore, futurechallenges remain regarding individual variations,morphological descriptions, haplotypes of host species
and surrounding environments including sediments.

components of detritus are recalcitrant polysaccharides
and bacteria are the main decomposers that degrade thesematerials. Therefore, we analyzed polysaccharide
degradation of the isolates. We found that many isolates
showed various polysaccharide degradation activities.
High diversity was observed in starch degradation
isolates, suggesting the large amount storage of starch in
detritus, for example algae. But, there were few isolates
showing alginate or agar degradation activities probably
because deep-sea was not suitable area for seaweeds
which contained a lot of alginate or agar. As mentionedin Fig.4, 11 species had no polysaccharides degradation
ability and 7 species of them in the anterior or mid partsof the intestine belonged to the family Bacillaceae 2 but
the number of them decreased in the posterior part of theintestine. It was observed that in the posterior part, thenumber of xylan degrading species related to the family
Bacilliaceae 1 increased and the number of starchdegrading species related to the order J-Proteobacteria
also increased (Fig.4). It was also observed that numberof species related to slight halophiles (3.5%) decreased in
the posterior part compared with those in other parts.
These results suggested that the posterior part had adifferent role or environment compared with the anterioror mid parts, maybe the posterior part was involved in
the digestion of polysaccharides.
We found that almost all isolates were facultative
anaerobic bacteria or anaerobic tolerant bacteria. It
appears that oxygen will enter the intestine of the sea
cucumber from the mouth with the detritus food and also
some amount can penetrate from the body tissues. These
results suggested that the aerobic culturable isolates
potentially contributed to digest detritus and supply
metabolic products (minor components and vitamins) totheir host sea cucumber.
Three isolates (isolate no. C214, C254 and C271)
showed less than 96% identities with any type strain
sequences and two of them were obtained from alkaline
plates and 10% NaCl. These results suggested that theintestines of deep-sea holothurians were still new
resource for new species.
The temperature of seawater at the water depth of thesampling point was estimated to be ca.13-14 °C in
November 2010 from data of Japan Meteorological
Agency. But, it was surprising that the full year
temperature of the sampling point was estimated to beca.12-15 °C throughout the year (from data in Japan
Meteorological Agency). These results suggested that
this deep-sea environment seemed a little lowtemperature but more suitable environment for deep-sea
holothurian and their intestinal bacteria.

Acknowledgments
Authors thank the captain and crews of TS Nagasaki
maru who made it possible to collect a deep-sea
cucumber. Authors are grateful for the support provided
by Dr. Asami Yoshida in Graduate School of Fisheries
Science and Environmental Studies, Nagasaki University
for DNA sequencing analysis. This work was partially
supported by a Grant-in-Aid for Scientific Research (C)to T. K.
References
1) Amaro, T., Luna, G.M., Danovaro, R., Billett,
D.S.M., and Cunha, M.R. 2012. High prokaryotic
biodiversity associated with gut contents of theholothurian Molpadia musculus from the Nazaré
Canyon (NE Atlantic). Deep-Sea Research I. 63:
82-90.
2) Amaro, T., Witte, H., Herndl, G.J., Cunha, M.R.,
and Billett, D.S.M. 2009. Deep-sea bacterial
communities in sediments and guts ofdeposit-feeding holothurians in Portuguese canyons(NE Atlantic). Deep-Sea Research I. 56:
1834-1843.
3) Enomoto, M., Nakagawa, S., and Sawabe, T. 2012.
Microbial
Communities
Associatedwith
Holothurians: Presence of Unique Bacteria in the
Coelomic Fluid. Microbes Environ., Advance
Publication, Release Date: March 23,2012.
4) Foster, G.G., and Hodgson, A.N. 1995. Annual
reproductive cycles of three sympatric species ofintertidal holothurians (Echinodermata) from thecoast of the Eastern Cape Province of South Africa.
Invertebr. Reprod. Dev. 27: 49-59.
5) Hagen, E.M., McCluney, K.E., Wyant, K.A.,
Soykan, C.U., Keller, A.C., Luttermoser, K.C.,
Holmes, E.J., Moore, J.C., Sabo, J.L. 2012. A
meta-analysis of the effects of detritus on primary
producers and consumers in marine, freshwater, andterrestrial ecosystems. Oikos, Article first published
online:
31
JAN
2012
|
DOI:
10.1111/j.1600-0706.2011.19666. x.
6) Massin, C. 1982. Food and feeding mechanisms,
Holothuroidea. In Echinoderm Nutrition, ed. by
Jangoux, M. and Lawrence, J.M., Balkema,
Rotterdam, pp. 43-55.
57

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)

7)

8)

9)

Moriarty, D.J.W. 1982. Feeding of Holothuria atraand Stichopus chloronotus on Bacteria, Organic
Carbon and Organic Nitrogen in Sediments of theGreat Barrier Reef. Aust. J. Mar. Freshwater Res.
33: 255-263.
Shimizu, M., Mikami, I., and Takahashi, K. 1992.
Histochemical detection on the ontogenic
development of digestive enzymes in the intestineof a juveni1e sea cucumber Stichopus japonicas.
Bulletin of the faculty of Fisheries Hokkaido
University 45: 1-8.
Uthicje, S., Schaffelke, B., and Byrne, M. 2009. A

boom and bust phylum? Ecological andevolutionary consequences of density variations inechinoderms. Ecological Monographs. 79: 3-24.
10) Ward-Rainey, N., Rainey, F.A., and Stackebrandt, E.
1996. A study of the bacterial flora associated with
Holothuria atra. J. Exp. Mar. Biol. Ecol. 203:
11-26.
11) Yingst, J.Y. 1976. The utilisation of organic matterin shallow marine sediments by an epibenthic
deposit-feeding Holothurian. J. Exp. Mar. Biol.
Ecol. 23: 55-69.

58

C210
C212
C214
C219
C220
C221
C234
C235
C236
C240
C241
C242
C243
C245
C246
C247
C254
C258
C259
C263
C265
C270
C271
C277
C280
C281
C285
C287
C288
C291
C295
C298
C302
C304
C305
C307
C309
C310
C312
C313
C317
C318
C324
C327
C333
C334
C335
C336
C339
C341
C344
C347

Isolate No.

AB741781
AB741782
AB741783
AB741784
AB741785
AB741786
AB741787
AB741788
AB741789
AB741790
AB741791
AB741792
AB741793
AB741794
AB741795
AB741796
AB741797
AB741798
AB741799
AB741800
AB741801
AB741802
AB741803
AB741804
AB741805
AB741806
AB741807
AB741808
AB741809
AB741810
AB741811
AB741812
AB741813
AB741814
AB741815
AB741816
AB741817
AB741818
AB741819
AB741820
AB741821
AB741822
AB741823
AB741824
AB741825
AB741826
AB741827
AB741828
AB741829
AB741830
AB741831
AB741832

Accession
number ofisolates

LB(pH7,3.5%)
LB(pH7,3.5%)
LB(pH7,3.5%)
LB(pH7,3.5%)
LB(pH7,3.5%)
LB(pH7,3.5%)
CMC(pH10,10%)
CMC(pH10,10%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
CMC(pH10,3.5%)
AL(pH10,10%)
AL(pH10,3.5%)
AL(pH10,3.5%)
AL(pH10,3.5%)
XL(pH10,10%)
XL(pH10,10%)
XL(pH10,10%)
XL(pH10,3.5%)
XL(pH10,3.5%)
XL(pH10,3.5%)
XL(pH10,3.5%)
XL(pH10,3.5%)
XL(pH10,3.5%)
XL(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
S(pH10,3.5%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
CMC(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)

isolation medium

528
818
623
825
819
509
817
813
594
499
589
540
797
812
574
427
453
621
715
524
535
565
764
677
544
475
581
637
772
521
515
816
411
540
683
550
565
623
516
388
580
780
569
811
679
586
808
740
589
558
768
704

letters

mid
mid
mid
mid
mid
mid
posteriorposterioranterior
mid
mid
mid
mid
mid
posteriorposterioranterior
anterior
anterior
posterioranterior
posteriorposterioranterior
mid
mid
posteriorposteriorposteriorposterioranterior
anterior
anterior
mid
mid
posteriorposteriorposteriorposteriorposterioranterior
anterior
mid
mid
posteriorposteriorposteriorposterioranterior
anterior
anterior
anterior

part of theintestine

AJ831844/AJ831842
AY543169
AB195680
AY543169
CP000002
AJ310149
AF483625
AJ640134
AJ831842/AJ831841
AJ831844/AJ831842
AY822043

EF114313
AF483624
AJ009793
AJ310149
L37603
AJ491290
HM054473
AB125942
AY190535
X76440D87035
AJ842344
D87035GU784860
AB188090
AJ492830
AJ717299
EU925618
AJ316187
AF057645
AB188090
HM054473
AJ605773
AB043865
D84025
X76444/AB292819
AY190535
HM054473/AB188090
X76449AB043851
DQ534014
HM054473
X74710
AJ842344
X76440AB043846
AJ842344
AF057645
AJ310647
AB043865
AB099708

Accession nnumber of typestrainBacillus aryabhattai
Bacillus marisflavi
Virgibacillus marismortui
Halobacillus trueperi
Staphylococcus warneri
Vibrio pomeroyi
Bacillus hunanensis
Marinobacter alkaliphilus
Jeotgalibacillus campisalis
Bacillus clausii
Bacillus horti
Photobacterium rosenbergii
Bacillus horti
Oceanobacillus kimchii
Bacillus oshimensis
Pseudomonas cedrina
Thalassobacillus devorans
Bacillus neizhouensis
Vibrio rotiferianus
Pseudomonas libanensis
Bacillus oshimensis
Bacillus hunanensis
Bacillus aurantiacus
Bacillus horikoshii
Pseudomonas synxantha
Bacillus clarkii/Bacillus polygoni
Jeotgalibacillus campisalis
Bacillus hunanensis/Bacillus oshimensis
Bacillus pseudalcaliphilus
Bacillus wakoensis
Photobacterium lutimaris
Bacillus hunanensis
Vibrio mediterranei
Photobacterium rosenbergii
Bacillus clausii
Bacillus hemicellulosilyticus
Photobacterium rosenbergii
Pseudomonas libanensis
Vibrio agarivorans
Bacillus horikoshii
Bacillus vietnamensis
Bacillus aerophilus/Bacillus altitudinis
Virgibacillus halodenitrificans
Halobacillus kuroshimensis
Virgibacillus halodenitrificans
Bacillus licheniformis
Halobacillus trueperi
Bacillus aquimaris
Oceanobacillus oncorhynchi
Bacillus altitudinis/Bacillus stratosphericus
Bacillus aerophilus/Bacillus altitudinis
Virgibacillus dokdonensis

species/tentative species

609/616 (98%)
691/699 (98%)
523/524 (99%)
530/535 (99%)
565/565 (100%)732/759 (96%), Gaps = 6/759 (0%)
675/677 (99%), Gaps = 1/677 (0%)
540/544 (99%)
474/475 (99%)
581/583 (99%), Gaps = 1/583 (0%)
634/637 (99%)
743/760 (97%)
521/521 (100%)
512/515 (99%)
815/816 (99%)
407/411 (99%)
535/540 (99%)
673/677 (99%)
547/550 (99%)
560/565 (99%)
619/623 (99%)
505/516 (97%)
388/388 (100%)
579/580 (99%)
780/780 (100%)568/569 (99%)
808/811 (99%), Gaps = 2/811 (0%)
675/676 (99%)
580/580 (100%)
806/808 (99%)
735/740 (99%)
577/589 (97%)
558/558 (100%)768/768 (100%)691/704 (98%)

522/522 (100%)
818/818 (100%)555/577 (96%)
816/825 (98%)
819/819 (100%)
503/509 (98%)
816/817 (99%)
807/813 (99%)
590/594 (99%)
497/499 (99%)
585/589 (99%), Gaps = 1/589 (0%)
537/540 (99%)
791/797 (99%), Gaps = 2/797 (0%)
807/812 (99%)
571/574 (99%)
425/427 (99%)
437/453 (96%)

Identities

+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
-

S

    

+
+
+
+
+
+
+
+
+
-

CMC
+
+
-

AL
+
+
+
+
+
+
+
-

XL

Degrading activities onpolysaacharides

FA
AT
AT
AT
AT
FA
AT
AT
AT
AT
AT
FA
AT
FA
FA
FA
AT
FA
FA
AT
AT
AT
AT
AT
FA
AT
FA
AT
FA
FA
FA
FA
FA
FA
FA
AT
FA
AT
FA
FA
FA
AT
AT
AT
AT
FA
AT
AT
AT
AT
AT
AT

Requimentof oxygen

10
20
15
20
15
3.5
20
15
3.5
15
3.5
3.5
3.5
20
15
15
20
15
3.5
10
15
15
20
10
10
15
10
15
10
15
3.5
15
3.5
10
10
10
10
15
10
10
10
10
20
25
20
15
20
15
20
10
10
10

NE
NE
ALK
ALK
NE
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
ALK
NE
NE
NE
NE
NE
NE
NE
NE
ALK
NE
NE
NE

MaximumpH
NaClconcentration tolerance
for growth
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
proteobacteria
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
firmicutes
proteobacteria
proteobacteria
firmicutes
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
proteobacteria
proteobacteria
firmicutes
firmicutes
proteobacteria
proteobacteria
proteobacteria
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes

phylumfamily Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family Bacillaceae 2family Staphylococcaceae
gamma
family Bacillaceae 1
gamma
family Planococcaceae
family Bacillaceae 1family Bacillaceae 1
gamma
family Bacillaceae 1family Bacillaceae 2family Bacillaceae 1
gamma
family Bacillaceae 2family Bacillaceae 1
gamma
gamma
family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1
gamma
family Bacillaceae 1family Planococcaceae
family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1
gamma
family Bacillaceae 1
gamma
gamma
family Bacillaceae 1family Bacillaceae 1
gamma
gamma
gamma
family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family Bacillaceae 2family Bacillaceae 2family Bacillaceae 1family Bacillaceae 2family Bacillaceae 1family Bacillaceae 2family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family

C350
C355
C358
C359
C360
C361
C362
C365
C367
C370
C375
C377
C379
C380
C381
C392
C397
C416
C417
C431
C435
C442
C444
C446
C460
C463
C466
C471
C475
C483
C484
C487
C494
C503
C506
C512
C520
C521
C528
C538

AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
AL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
XL(pH7,10%)
S(pH7,10%)
S(pH7,10%)
S(pH7,10%)
CMC(pH7,3.5%)
CMC(pH7,3.5%)
CMC(pH7,3.5%)
CMC(pH7,3.5%)
AL(pH7,3.5%)
AL(pH7,3.5%)
AL(pH7,3.5%)
XL(pH7,3.5%)
XL(pH7,3.5%)
XL(pH7,3.5%)
XL(pH7,3.5%)
XL(pH7,3.5%)
XL(pH7,3.5%)
S(pH7,3.5%)
S(pH7,3.5%)
S(pH7,3.5%)
S(pH7,3.5%)
S(pH7,3.5%)
S(pH7,3.5%)
S(pH7,10%)

823
684
753
528
557
665
813
772
520
420
598
760
533
535
836
655
568
711
672
777
706
531
421
568
477
462
664
544
655
673
542
535
761
749
540
556
563
669
578
746

mid
mid
posteriorposteriorposteriorposteriorposteriorposteriorposterioranterior
anterior
anterior
mid
mid
mid
posteriorposteriormidposteriorposterioranterior
mid
mid
mid
anterior
midposterioranterior
anterior
mid
mid
mid
posterioranterior
anterior
mid
mid
mid
posteriorposterior

AF057645
AJ306891

AB195680
AB195680
AF541966
CP000002
AB099708
AY543169
AB195680
AF483625
AF541966
AB101591
AB473561
AB195680
CP000002
X76440AB195680
CP000002
AJ842344
CP000002
CP000002
AY543169
AJ491290
AJ491290
D16273
AY876289
AJ491290
AF057645
AJ316187
AF057645
AJ316187
AF057645
AJ492830
AY750575
AJ831844
D16273
AB021185
AF057645
AJ492830
AY793550

Halobacillus kuroshimensis
Halobacillus kuroshimensis
Bacillus hwajinpoensis
Bacillus licheniformis
Bacillus vietnamensis
Virgibacillus halodenitrificans
Halobacillus kuroshimensis
Bacillus aquimaris
Bacillus hwajinpoensis
Gracilibacillus dipsosauri
Oceanobacillus sojae
Halobacillus kuroshimensis
Bacillus licheniformis
Bacillus clausii
Halobacillus kuroshimensis
Bacillus licheniformis
Photobacterium rosenbergii
Bacillus licheniformis
Bacillus licheniformis
Virgibacillus halodenitrificans
Vibrio pomeroyi
Vibrio pomeroyi
Bacillus megaterium
Bacillus pumilus
Vibrio pomeroyi
Pseudomonas libanensis
Vibrio rotiferianus
Pseudomonas libanensis
Vibrio rotiferianus
Pseudomonas libanensis
Pseudomonas cedrina
Vibrio harveyi
Bacillus aerophilus
Bacillus megaterium
Bacillus flexus
Pseudomonas libanensis
Pseudomonas cedrina
Bacillus lehensis
Pseudomonas libanensis
Halomonas meridiana

816/820 (99%), Gaps = 3/820 (0%)
682/685 (99%), Gaps = 2/685 (0%)
746/749 (99%), Gaps = 1/749 (0%)
528/528 (100%)557/557 (100%)663/665 (99%)
810/814 (99%), Gaps = 2/814 (0%)
760/772 (98%)
515/520 (99%)
420/420 (100%)
596/598 (99%)
758/761 (99%), Gaps = 2/761 (0%)
531/532 (99%)
534/535 (99%)
831/837 (99%), Gaps = 2/837 (0%)
653/655 (99%)
565/569 (99%), Gaps = 1/569 (0%)
710/711 (99%)
671/672 (99%)
777/777 (100%)699/706 (99%), Gaps = 1/706 (0%)
525/531 (98%)
417/421 (99%)
568/569 (99%)
471/477 (98%)
461/462 (99%)
662/664 (99%)
542/544 (99%)
654/655 (99%)
671/673 (99%)
540/542 (99%)
529/535 (98%)
761/761 (100%)
747/749 (99%)
538/540 (99%)
554/556 (99%)
561/564 (99%), Gaps = 1/564 (0%)
668/669 (99%)
576/578 (99%)
746/746 (100%)

+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+

+
+
+
+
+
+
+
+
+
+
-

+
-

+
+
+
+
+
+
+
+
-

A
AT
AT
AT
AT
AT
A
AT
AT
AT
AT
FA
FA
AT
A
FA
FA
FA
FA
AT
FA
FA
AT
FA
FA
AT
FA
FA
FA
AT
FA
FA
AT
FA
AT
FA
FA
AT
FA
AT

20
20
15
10
10
20
25
15
20
15
20
25
15
25
20
15
15
15
15
20
3.5
3.5
10
10
3.5
10
3.5
15
15
15
15
15
15
10
15
10
15
15
10
15

NE
NE
NE
NE
NE
NE
NE
NE
NE
ALK
ALK
ALK
ALK
NE
ALK
NE
NE
NE
NE
NE
ALK
ALK
NE
NE
ALK
ALK
ALK
NE
ALK
ALK
ALK
ALK
NE
NE
NE
NE
NE
ALK
NE
ALK

firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
firmicutes
proteobacteria
firmicutes
firmicutes
firmicutes
proteobacteria
proteobacteria
firmicutes
firmicutes
proteobacteria
proteobacteria
proteobacteria
proteobacteria
proteobacteria
proteobacteria
proteobacteria
proteobacteria
firmicutes
firmicutes
firmicutes
proteobacteria
proteobacteria
firmicutes
proteobacteria
proteobacteria

family Bacillaceae 2family Bacillaceae 2family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family Bacillaceae 2family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family Bacillaceae 2family Bacillaceae 2family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2family Bacillaceae 1
gamma
family Bacillaceae 1family Bacillaceae 1family Bacillaceae 2
gamma
gamma
family Bacillaceae 1family Bacillaceae 1
gamma
gamma
gamma
gamma
gamma
gamma
gamma
gamma
family Bacillaceae 1family Bacillaceae 1family Bacillaceae 1
gamma
gamma
family Bacillaceae 1
gamma
gamma

Abbreviations: S, starch; CMC, carboxymethyl cellulose sodium salt; AL, alginate; XL, xylan; FA, facultative anaerobic bacteria; AT, anaerobic tolerant bacteria; A, aerobic bacteria;NE, neutrophilic bacteria; ALK, alkaliphilic/alkali-tolerant bacteria . Display of more than one species in the column of nearest
type strain species indicated the same idenntity in the comparison range.

AB741833
AB741834
AB741835
AB741836
AB741837
AB741838
AB741839
AB741840
AB741841
AB741842
AB741843
AB741844
AB741845
AB741846
AB741847
AB741848
AB741849
AB741850
AB741851
AB741852
AB741853
AB741854
AB741855
AB741856
AB741858
AB741859
AB741860
AB741861
AB741862
AB741863
AB741864
AB741865
AB741866
AB741867
AB741868
AB741869
AB741870
AB741871
AB741872
AB741873

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪦㪩㪠㪞㪠㪥㪘㪣㩷㪧㪘㪧㪜㪩㩷㪥㪦㪫㪜㩷Journal of Japanese Society for Extremophiles (2012) Vol.11, 59-63

Ueno Y, Kato-Yamada Y and Imai T

Hydrophobic shield on the molecular surface enhances thermal
stability of ferredoxin of Cyanidioschyzon merolae
Department of Life Science, College of Science, Rikkyo University, Nishi-Ikebukuro Toshima-ku, Tokyo 171-8501,
Japan
Correspondence author: Yuko Ueno, y.ueno@rikkyo.ac.jp
Tel: +81-3-3985-2386
Fax: +81-3-3985-2386
Received: Nobember 8, 2012 / Accepted: November 19, 2012
One of the most generally recognized factors forenhancing the protein thermal stability is to increase thepacking density of the hydrophobic core5,12). In addition,amino acid substitution at the surface has been shown tohave little effect on protein thermal stability10,17).
However, examples that few point mutations at thesurface drastically increase the proteins thermal stability
have been also reported15,19). The factors that enhance theprotein’s thermalstablity seem to vary with regard to theproteins to be focused on and general concepts to raise
the thermal stability are still a matter of debate.
Cyanidioschyzon merolae (Cm) is a moderately
thermophilic red alga, whose optimum growthtemperature is 42°C–50°C. It is also a uniquethermophile that is positioned as a primitive eukaryote
cell according to its simple organelle structure11).
Ferredoxin (Fd) is an iron-sulfur protein present in manylife forms where it acts as a multifunctional electron
carrier9,18). The -grasp motif is a protein motif that
forms the core of Fd with five -strands and one
-helix2,8,14). This motif is also present in ubiquitin, oneof the most phylogenetically well-conserved proteins ineukaryotes and several other proteins with biologicallydistinct functions. In many studies of thermostability ofproteins, more thermophilic or hyperthermophilicproteins are used because of their attractive features.
However, we chose to compare a moderately
thermophilic protein with the mesophilic protein from
Spinacia oleracea (So)1) because it has more similarities
in its primary, secondary, and tertiary structures, and has
still approximately 20°C difference in thermal stability20),
which may unobtrusively carries a unified set of rules forprotein thermal stability.

Abstract Ferredoxin from Cyanidioschyzon merolae, amoderately thermophilic unicellular alga, has ahydrophobic interaction between two solvent-exposed
residues, V53 and V87, which are specific forthermophilic ferredoxins. This interaction is located ontwo -strands, which are part of a -grasp motif
constructing ferredoxin. Point mutations were designed
to those two residues, which were participating in thissurface hydrophobic interaction, and it was revealed thatthe thermal stability of the V87A mutant was largely
decreased compared to that of the wild type (WT). The
result suggested an example for the finding that surfacehydrophobicity affected protein thermal stability.
Key words: Ferredoxin; thermal stability, hydrophobicity, -grasp motif, Cyanidioschyzon merolae
Abbreviations
Cm:
Cyanidioschyzon merolae
So:
Spinacia oleracea
Ml:
Mastigocladus laminosus
Fd:
Ferredoxin
WT:
Wild typeCmFd: Cyanidioschyzon merolae ferredoxin
SoFd:
Spinacia oleracea ferredoxin
MlFd:
Mastigocladus laminosus ferredoxin
SoFd-NADP reductase:
Spinacia oleracea ferredoxin nicotinamide adenine
dinucleotide phosphate (oxidized form) reductase
NADPH: Nicotinamide adenine dinucleotide phosphate(reduced form)

59

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)After the proteins had been applied to the first twocolumns, ammonium sulfate was added to the eluate to a
final concentration of 2 M, and the solution was loaded
onto a DEAE Sepharose FF column (GE Healthcare). Fd
was eluted with a negative linear gradient of 2-0 M
ammonium sulfate and the purity was confirmed bynative polyacrylamide gel electrophoresis. The Fd
concentration was estimated on the basis of its
absorption at 420 nm, using an extinction coefficient of
9.68 mM-1 cm-1 18).
For the measurement of protein thermal stability, Fd
samples were assayed for cytochrome c reductase
electron transfer activity13,20). Fd samples were incubatedat each temperature (ranging between 20°C and 80°C)
for 30 min. Continuously, the incubated Fd samples wereadded to an assay mixture that consisted of 50 mM
Tris-HCl (pH 8.0), 250 μM NADPH, 39 μM bovine
heart cytochrome c, and 3.3 U/mL SoFd-NADP
reductase. Immediately thereafter, cytochrome creduction was monitored spectrophotometrically at 550
nm at 25°C. The temperature midpoint of the remaining
electron transfer activity of Fd was calculated by fittingwith the Boltzmann function.
The hydrophobic core of Fd is located in the innerspace of the -grasp motif, which is formed by five-strands and one -helix (Fig. 1). In addition, there is ahydrophobic interaction between the side chains on thesurface of CmFd, i.e., V53 on 3 and V87 on 5 (Fig. 1).

By comparing the structure of the plant-type [2Fe-2S]
ferredoxin from Cm and So (a mesophilic higher-plant),
we found that CmFd has a specific hydrophobic
interaction occurs on the surface of the protein asbuilding up a hydrophobic shield that is likely to protectthe -sheet structure. In this study, by introducing point
mutations into this region, we examined the effect of
surface hydrophobic interactions on the protein thermal
stability.
All mutants were prepared by inverse polymerase chain
reaction with the KOD-Plus-Mutagenesis Kit(TOYOBO). We used the pTTQ18 vector (Amersham
Pharmacia Biotech) that carries the WT Fd gene as atemplate. The V87A mutant was prepared by using two
oligonucleotide primers, i.e., forward primer (bold letters
are the mutational points):
5-GCCATTCAAACTCATCAAGAAGAAGCT-3, andreverse primer: 5-GCAATCAGAAGTAGGATATGCAACACAGGTTAG-3. The sequences of the V87T
mutant primers were as follows: forward 5-GTACTATTCAAACTCATCAAGAAGAAGC-3, reverse 5-AATCAGAAGTAGGATATGCAACACAGG-3. Those of the
V53K mutant primers were as follows: forward
5-AAGAAAGGATCCGTGGATCAATCTGATC-3,
reverse 5-TAATTTGCCTGCGCAAGTAGAGCAAG3. The linear DNA fragment containing the mutation
was ligated into circular DNA and the mutant proteins
were expressed and purified as described previously20).

Fig. 1. Crystal structure of CmFd WT(a) The [2Fe-2S] cluster is shown in stick representation. Fe and S atoms are orange and yellow, respectively. “N” and“C” stand for N-terminal and C-terminal end, and the secondary structure elements are represented. Five -strands,1–5, and an -helix, 1, form the -grasp motif, and the hydrophobic core exists in the interior of this motif.
(b) (a) viewed from a different angle. V53 and V87, shown in stick representation, located on 3 and 5, respectively.
The side chains are exposed to the surface of the molecule.

60

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Thermophilic cyanobacterium Mastigocladus laminosus
Fd (MlFd; optimum activity at 65°C6)) also has
hydrophobic residues at this position, but SoFd has K52
and T86 at the corresponding site. Therefore, we
designed a point mutation in CmFd that replaces V87
with Ala and assayed protein thermal stability bymeasuring Fd electron transfer activity. The midpointtemperature of the remaining activity of V87A was 61°C,
which was largely decreased compared to 71°C of the
CmFd (Fig. 2). This result may indicate that thehydrophobic interaction in this region is indeed

important for protein thermal stability. With regard to the
V87T mutant (amino acid substitution corresponding toSoFd), the midpoint temperature was 67°C, which is not
as low as that of V87A, but still showed a decrease inprotein stability. With regard to the V53K mutant, inwhich an amino acid was substituted that was holding ahydrophobic interaction with V87 and corresponded to
SoFd, the midpoint temperature was 66°C. The double
mutant V53K/V87T, in which both V53 and V87 were
replaced with residues corresponding to SoFd, had a
midpoint temperature of 67°C.

Fig. 2. Measurement of Fds thermal stability
Electron transfer activity of Fd remain after the incubation with indicated temperature was monitored in Fd-NADP+
reductase system by absorption at 550 nm for reduction of cytochrome c. Symbols represent, CmFd WT; closed circles,
SoFd; closed squares, CmFd V87A mutant; closed triangles, CmFd V87T mutant; open circles, CmFd V53K mutant;open squares, CmFd V53K/V87T double mutant; open triangles. Lines represent fitting with the Boltzmann function.

Comparison of the tertiary structure of CmFd and SoFd
showed that CmFd has a specific hydrophobic region
formed by V53 and V87 and -CH2- from K51, L54, andQ89 (Fig. 3). On the other hand, SoFd does not havehydrophobic residues but has an ion pair between K52
and E88 (corresponds to V53 and Q89 in CmFd,
respectively) instead in this region. It is generally
assumed that close packing of the hydrophobic core andminimized cavities play a major role in protein
stability3,5,12), while amino acid substitution on thesurface of the molecular has little effect on its
stability10,17). Yet, exceptions for these general rules havebeen reported by demonstrating the few point mutations
at the surface resulted in a sufficient increase in protein
stability15,16,19). Since both CmFd and SoFd have ahydrophobic core under the -sheet containing these two-strands, a hydrophobic shield on this region may be

important for the structure and thermal stability of Fd.
Indeed, MlFd has this hydrophobic shield at this region,
too.
Among all 4 mutants, the decrease in thermal stability
was the highest for the V87A mutant. It is most favorable
to think that this was caused by a loss in the volume ofthe hydrophobic group. The loss of the methyl group
might create a hole in the hydrophobic shield, andthereby destabilized this region. MlFd, which has higher
thermal stability, has Ile instead of Val in this region (Fig.
3). A closer packed hydrophobic shield may enablehigher protein stability. In contrast, among 3 mutants inwhich the residues were replaced according to the one in
SoFd, V87T, V53K, and V53K/V87T, there was noremarkable difference in their thermal stability. This
result indicated the importance of “inter -strands” actionof hydrophobic residues. Furthermore, it indicated that61

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)hydrophobic -CH2- parts in Thr and especially Lys4).
These methylene groups covered the lack in thehydrophobic shield and, as a result, the mutants did notlose their thermal stability as did the substitution to Ala.

substitution of one amino acid had the same effect as thatof two with regard to hydrophobic interactions. However,these 3 mutants, in which Val was substituted with Lys
and Thr, did not lose its thermal stability as did in thecase of V87A. This might be due to the volume of the

T53

T86

K52
3

V87

K54
V53

4

V288

S255
I254

4
3

4
3

Q89

E88
K50

E290

K252

K51
K91

K293
E296

E71

SoFd

CmFd

MlFd

Fig. 3. Surface region of 3 and 5 in 3 different Fds
Five residues that face the surface of the molecule in the 3 and 5 regions from SoFd (blue), CmFd (pink), MlFd
(orange) are shown in ball and stick representation. Yellow dashed lines represent the ion pairs. The light-blue circle
represents the hydrophobic region. CmFd and MlFd both have a hydrophobic region that shields the  sheet. SoFd does
not have hydrophobic residues, but has an ion pair instead.

of mesophilic ferredoxin by introducing a hydrophobic
shield should be examined to confirm the hypothesis.
The examples of hydrophobic shields protecting the-sheet structure against water molecules were also
reported for thermophilic proteases and cold shockproteins7,16). In addition, in CmFd, the existence of ahydrophobic shield on the surface of the -sheet in the-grasp motif represents the importance of this region
and may be the key point for investigating strategies forprotein thermal stability. We are further performing
structure-based analyses of CmFd and its mutants andinvestigating its mechanism of thermal stability.

SoFd does not possess the hydrophobic shield but instead
has an ion pair in this region. Since ion pairs haverelatively high binding energy, it may be generally
thought that this could contribute more to theenhancement of protein stability. Nevertheless, CmFd
and MlFd, which have hydrophobic residues in thissurface -sheet region, had higher thermal stability.
Additionally, one amino acid substitution in this region
can decrease its protein stability by 10°C compared to
the CmFd. This demonstrates that shielding the -sheetsfrom solvents (in many cases water) by hydrophobic
residues is more important than to connect the two-sheets with a strong bond. Placing the hydrophobic
shield on the surface of -strands presumably enabled theproteins to obtain a higher thermal stability by
preventing an invasion of water molecules into the coreof the protein. However, increasing the thermal stability

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62

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Journal of Japanese Society for Extremophiles (2012) Vol.11, 64-69

Tsuda S.a,b

Antifreeze proteins from Japanese fish࿖↥㝼㘃↱᧪ߩਇಓ࠲ࡦࡄࠢ⾰↥ᬺᛛⴚ✚ว⎇ⓥᚲ↢‛ࡊࡠ࠮ࠬ⎇ⓥㇱ㐷ർᶏ㆏ᄢቇ↢๮⑼ቇ㒮ᵤ↰ ᩕaBioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1
Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, JAPAN.
b
Graduate school of life science, Hokkaido University
Corresponding author: Sakae Tsuda, s.tsuda@aist.go.jp
Phone: +81-11-857-8912, Fax: +81-11-857-8983
Received: October 26, 2012 / Accepted: November 16, 2012ߪߓ߼ߦ ߎࠇ߹ߢߦ‫ޔ‬ਇಓ࠲ࡦࡄࠢ⾰㧔Antifreeze Protein:
AFP‫ޔ‬ਇಓ♧Ⱞ⊕⾰ AFGP ࠍ฽߻㧕ߪ㝼㘃ߩߺߥࠄߕ᣸⯻‫ޔ‬ᬀ‛‫⩶ޔ‬㘃ߥߤ߆ࠄ߽⊒⷗ߐࠇ‫ߪࠄࠇߘޔ‬10 ⒳㘃એ਄ߩ⇣ߥࠆ࠲ࠗࡊߦᄢ೎ߐࠇߡ޿ࠆ‫ࠇߘޕ‬ࠄߩ AFP ߪฦ‫␲ߩޘ‬వⰮ⊕⾰߇⁛┙ߦಽሶㅴൻࠍ
ㆀߍߡ᳖⚿᥏⚿ว⢻ജࠍ₪ᓧߔࠆߦ⥋ߞߚߣ⠨߃ࠄࠇߡ߅ࠅ‫␲ޔ‬వⰮ⊕⾰ߩᐞߟ߆ߪᣢߦ․ቯߐࠇߡ޿
ࠆ‫ޔߜࠊߥߔޕ‬୘‫ ߩޘ‬AFP ߩ㑆ߢߪࠕࡒࡁ㉄㈩೉‫ޔ‬ಽሶ㊂‫ޔ‬㧟ᰴరಽሶ᭴ㅧ‫ޔ‬᳖⚿᥏⚿วㇱ૏‫ޔ‬ᾲ቟ቯᕈ‫ޔ‬ਇಓᵴᕈ(ᾲࡅࠬ࠹࡝ࠪࠬ)╬߇ᭂ߼ߡᄢ߈ߊ⇣ߥߞߡ޿ࠆ 2,6)‫ޔ߼ߚߩߎޕ‬AFP ߩታ↪ൻࠍ⠨߃ࠆ㓙ߦߪ‫ޔ‬ᢙ޽ࠆ AFP ߩਛߢߤߩ⒳㘃߇૗ߩ↪ㅜߦ૶߃ࠆߩ߆ࠍ♖ᩏߒ‫ߦࠄߐޔ‬ᵈ⋡ߔࠆ AFP ߇቟ଔߦᄢ
㊂↢↥ߢ߈ࠆ߆ุ߆ࠍ᣿ࠄ߆ߦߔࠆ੐߇㕖Ᏹߦ㊀ⷐ
ߦߥࠆ‫ޕ‬᳖⚿᥏⚿วᯏ⢻ߣ⚦⢩⤑଻⼔ᯏ⢻ࠍ૬ߖᜬߟ㝼㘃↱᧪ߩ AFP㧔࿑㧝㧕ߪ‫ޔ‬᳓↥⾗Ḯᄢ࿖࡮ᣣᧄ
ߩ․ᓽࠍ↢߆ߔ᦭↪ᯏ⢻ᕈ‛⾰ߩ৻ߟߦߥߞߡ޿ߊ
น⢻ᕈ߇޽ࠆ‫ޕ‬

Abstract Antifreeze protein (AFP) is an extraordinary
bio-molecule that specifically binds to the surface of anice crystal to inhibit its growth below 0 C, and also to
the membrane surface of a cell to improve its viability
above 0C. Based on our knowledge about thebiochemical/biophysical properties as well as the
structure-function relationships of various AFPs, we aretrying to develop new AFP-technologies applicable fromindustry to medical fields.
Keywords: antifreeze protein, ice-binding, ice
recrystallization inhibition, three-dimensional structure.
ࠠ࡯ࡢ࡯࠼㧦ਇಓⰮ⊕⾰‫ޔ‬᳖⚿᥏⚿ว‫ޔ‬ౣ⚿᥏㒖ኂ‫ޔ‬㧟ᰴర᭴ㅧⷐ⚂ਇಓ࠲ࡦࡄࠢ⾰㧔AFP㧕ߪ‫ޔ‬ಓ⚿ߔࠆ⋥೨ߩ᳓ߩౝ
ㇱߦήᢙߦ↢ᚑߔࠆ᳖ߩන⚿᥏㧔=᳖ᩭ㧕ߦኻߒߡ․⇣⊛ߦ⚿วߔࠆᯏ⢻㧔᳖⚿᥏⚿วᯏ⢻㧕ࠍ᦭ߔࠆ‫ޕ‬ߐࠄߦ‫ޔ‬㝼㘃↱᧪ߩ AFP ߪ᳖ߩߺߥࠄߕ⢽⾰ੑ㊀⤑ߦ߽ᒝߊ⚿วߒߡߘߩ⚦⢩ߩኼ๮ࠍᑧ߫ߔᯏ⢻㧔⚦⢩଻⼔ᯏ⢻㧕ࠍ⊒ើߔࠆ‫⪺ޕ‬⠪ࠄߪ‫ߦߢ߹ࠇߎޔ‬ᄢ㊂↢↥߇น⢻ߦߥߞߚ㝼㘃↱᧪ AFP ࠍᵴ↪ߔࠆߎߣߢᣂߚߥᛛⴚࠍ૞ࠅߚ޿ߣ⠨߃ߡ޿ࠆ‫ޕ‬㝼㘃↱᧪ AFP ߩ⒳㘃
 AFGP ߪ‫ ⚂ࠄ߆੹ޔ‬40 ᐕ೨ߦධᭂᶏߦ↢ᕷߔࠆࡁ
࠻࠮࠾ࠕ⑼㝼㘃ߩⴊᶧ߆ࠄ⷗಴ߐࠇߚೋ߼ߡߩ AFPߢ޽ࠆ‫ߩߎޕ‬ಽሶߪ Ala-Ala-Thr ߩ 3 ᱷၮࠍන૏ߣߒ
ߚ෻ᓳ㈩೉߆ࠄ᭴ᚑߐࠇ‫ޔ‬એਅߩࠃ߁ߥಽሶ㊂ߩ⇣ߥࠆ㧤୘ߩࠕࠗ࠰ࡈࠜ࡯ࡓߣߒߡ㝼૕ౝߦ⊒⃻ߐࠇ
ߡ޿ࠆ㧦AFGP1㧔34 kDa㧕‫ޔ‬AFGP2㧔29 kDa㧕‫ޔ‬AFGP3
㧔21.5 kDa㧕‫ޔ‬AFGP4㧔17 kDa㧕‫ޔ‬AFGP5㧔10.5 kDa㧕‫ޔ‬64

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)AFGP6㧔7.9 kDa㧕‫ޔ‬AFGP7㧔3.8 kDa㧕‫ޔ‬AFGP8㧔2.6 kDa㧕15)‫ޕ‬Thr ߩ஥㎮ߩ-OH ၮߦ⚿วߒߡ޿ࠆ 2 ♧(ࠟ࡜ࠢ࠻ࠪ࡞-N ࠕ࠮ ࠴࡞ࠟ࡜ࠢ࠻ࠨࡒࡦ)߇ਇಓᵴᕈ
ߦᔅ㗇ߣߐࠇߡ޿ࠆ‫ޕ‬AFGP ߪᭂ߼ߡ㜞޿ᾲ቟ቯᕈ
ࠍ᦭ߒߡ߅ࠅ‫ޔ‬90-95qC ߩᾲಣℂᓟߦ߅޿ߡ߽ਇಓ
ᵴᕈߩૐਅ߇߶ߣࠎߤߥ޿‫ޕ‬ൻቇวᚑߦࠃߞߡ⺞⵾ߒߚ AFGP ߩ෻ᓳන૏ߩ᭴ㅧ߇ NMR ᴺߦࠃߞߡ⸃ᨆߐࠇ 16)‫ߩߘޔ‬㛽ᩰ᭴ㅧߪ II ဳߩࡐ࡝ࡊࡠ࡝ࡦ
᭴ㅧߦ㘃ૃߒߚ 3 ᱷၮߏߣߦ৻๟ߔࠆᏀᏎ߈ࡋ࡝࠶
ࠢࠬ᭴ㅧߢ޽ࠆߎߣ߇᣿ࠄ߆ߦߥߞߚ‫♧ޕ‬㎮߇⚿วߒߡ޿ࠆ Thr ᱷၮߪࡋ࡝࠶ࠢࠬߩ ஥ߦ⋥✢⁁ߦ㈩⟎ߐࠇߡ߅ࠅ‫♧ޔ‬㎮ߦ޽ࠆ᳓㉄ၮߩᄙߊߪࡋ࡝࠶
ࠢࠬߦᐔⴕߦਗࠎߢ޿ߚ‫┙ߩࠄࠇߎޕ‬૕᭴ㅧ਄ߩ․ᓽ߇᳖⚿᥏ߣߩ⋧੕૞↪߇ AFGP ߩਇಓᵴᕈࠍ߽
ߚࠄߔߣ⠨߃ࠄࠇߡ޿ࠆ‫ޕ‬
 AFPI ߪ‫ޔ‬ධᭂᶏߢߪߥߊർᄢ⷏ᵗߩർㇱᴪጯߦ↢ᕷߔࠆࡈ࡙ࡅ࡜ࡔߩⴊᶧ߆ࠄ♧㎮ୃ㘼ࠍฃߌߡ޿ߥ
޿ೋ߼ߡߩ AFP ߣߒߡ⊒⷗ߐࠇߚ 1)‫ޕ‬AFPI ߪ‫ࠕޔ‬࡜࠾ࡦ߇᭴ᚑࠕࡒࡁ㉄ߩ 6 ഀએ਄ࠍභ߼ࠆන㎮ࡐ
࡝
ࡍ
ࡊ
࠴
࠼
ߢ
޽
ࠅ
‫ޔ‬
Thr-Ala-Ala-X-Ala-X-X-Ala-Ala-X-X(X ߪછᗧ) ߆ࠄ
ߥࠆ 11 ᱷၮߩࡕ࠴࡯ࡈߩ 3ޯ6 ࿁ߩ➅ࠅ㄰ߒ߆ࠄ᭴
ᚑߐࠇࠆ‫ ߜࠊߥߔޕ‬AFPI ߩᄤὼ‛߽ࠕࡒࡁ㉄⚵ᚑߣ㎮㐳ߩ⇣ߥࠆⶄᢙߩࠕࠗ࠰ࡈࠜ࡯ࡓߩᷙว‛ߢ޽ࠆ߇‫ߩߘޔ‬ಽሶ㊂ಽᏓߪ 3ޯ4.5 kDa ߣ㕖Ᏹߦ⁜޿▸࿐ߦ޽ࠆ‫ޔ߼ߚߩߎޕ‬AFGP ߦᲧߴࠆߣ㆔߆ߦኈᤃߦ̌ࠕࠗ࠰ࡈࠜ࡯ࡓᷙว‛̍ߣߒߡߩ AFPI ᄤὼ‛ࠍ㜞⚐ᐲߦ♖⵾ߔࠆߎߣ߇ߢ߈ࠆ‫․ࠆ޽ޕ‬ቯߩ AFPIࠕࠗ࠰ࡈࠜ࡯ࡓߦߟ޿ߡ⸃߆ࠇߚ X ✢᭴ㅧߪ‫ޔ‬㐳ߐ⚂ 55Έߩ 1 ᧄ㎮ǩ-ࠄߖࠎߢ޽ߞߚ‫ߪࠎߖࠄߩߎޕ‬3.6 ᱷၮߢ 1 ࿁ォߔࠆߚ߼(1 ᱷၮ޽ߚࠅ 100q࿁ォ)‫ޔ‬
11 ᱷၮߢߪ 1,100 ᐲߔߥࠊߜ⚂ 3 ࿁ォߔࠆ‫ߩߎޕ‬
᭴ㅧߩ․ᓽ߇ࡕ࠴࡯ࡈౝߩฦࠕࡒࡁ㉄ࠍࠄߖࠎゲߦኻߒߡᐔⴕ߆ߟ╬㑆㓒ߦ㈩⟎ߐߖߡ޿ࠆ‫ޕ‬11 ᱷၮᲤߦ޽ࠆ Thr ᱷၮห჻ߩ〒㔌ߪ 16.7Έߢ޽ࠅ‫ࡒ࡜ࡇޔ‬࠳࡞㕙ߣ๭߫ࠇࠆ᳖⚿᥏⴫㕙ࠍ᭴ᚑߔࠆ᳓ಽሶߩ㑆㓒(16.5Έ)ߣ߶߷╬ߒ޿‫ޔ߼ߚߩߎޕ‬ೋ߼ߪ AFPI ߩ
Thr ᱷၮߩ OH ၮ߇หಽሶࠍ᳖⚿᥏ߦ⚿วߐߖࠆㇱ૏ߢ޽ࠆߣ੍᷹ߐࠇߚ‫ߩߘޕ‬ᓟ‫ޔ‬Thr ஥㎮ߩࡔ࠴࡞ၮ߿ Ala ஥㎮ߩ⇹᳓ᕈ߇᳖⚿᥏㕙ߣߩ⋧੕૞↪ߦ㑐ਈߔࠆߎߣࠍᒝߊ␜ໂߔࠆታ㛎⚿ᨐ߇ᓧࠄࠇߚߚ߼‫ޔ‬
AFPI ߩ᳖⚿᥏⚿วࡔࠞ࠾࠭ࡓߦ㑐ߔࠆ⼏⺰ߪ⃻࿷߽⛯޿ߡ޿ࠆ 5)‫ޕ‬ ಽሶ㊂⚂ 13kDa ߩ AFPII ߪ AFGP ߿ AFPI ߩࠃ߁ߥ➅ࠅ㄰ߒ㈩೉ࠍ߽ߚߥ޿‫ޕ‬หಽሶਛߦߪ 10 ୘ߩ
Cys ᱷၮ߇ሽ࿷ߒ‫߇ߡߴߔߩߘޔ‬ಽሶౝ S-S ᨞ᯅࠍᒻᚑߒߡ޿ࠆ‫㉄ࡁࡒࠕޕ‬㈩೉‫ޔ‬Cys ߩ૏⟎‫┙ޔ‬૕᭴ㅧߦ㑐ߒߡࠞ࡞ࠪ࠙ࡓ⚿วဳ(C ဳ)࡟ࠢ࠴ࡦߩ৻ㇱ࠼ࡔࠗࡦߣߩ㑆ߦ㜞޿⋧หᕈ߇ߺࠄࠇࠆ‫ࡅޔߚ߹ޕ‬࠻↱᧪ߩࠞ࡞ࠪ࠙ࡓ⚿᥏⚿วⰮ⊕⾰(࡝࠰ࠬ࠲ࠪࡦ)ߣ߽⋧หᕈࠍ␜ߔ‫ޕ‬AFPII ߪਇಓᵴᕈߩ⊒⃻ߦ ࠞ࡞65ࠪ࠙ࡓࠍᔅⷐߣߒߥ޿߽ߩߣᔅⷐߣߔࠆ߽ߩߩ㧞⒳㘃ߦᄢ೎ߐࠇࠆ‫ޕ‬AFPII ᄤὼ‛߽ࠕࠗ࠰ࡈࠜ࡯ࡓߩ
ᷙว‛ߢߪ޽ࠆ߇‫ߩߘޔ‬⒳㘃ߪ㕖Ᏹߦዋߥ޿‫⪺ޕ‬⠪ࠄߪർᶏ㆏ߩࠝࡎ࡯࠷ࠢᶏᴪጯ╬ߦ↢ᕷߔࠆࠪ࠴ࡠ
࠙࠙ࠝ߆ࠄන㔌♖⵾ߐࠇߚࠞ࡞ࠪ࠙ࡓ㕖ଐሽဳߩII ဳ AFP ߩ X ✢᭴ㅧࠍ 2008 ᐕߦ᣿ࠄ߆ߦߒߚ 14)‫ޕ‬ߎߩಽሶߩਛᄩߦߪߨߓࠇߚㅒᐔⴕǪ-ࠪ࡯࠻߇޽
ࠅߘߩਔ஥ߦǩ-ࠄߖࠎ߇৻ߟߕߟ㈩⟎ߒߡ޿ߚ‫ޕ‬ห
AFPII ߩᄌ⇣૕૞ᚑታ㛎߆ࠄ‫ޔ‬AFPII ߩ᳖⚿᥏⚿วㇱ૏ߪǪ-ࠪ࡯࠻ߣߘߩㄭறߩ࡞࡯ࡊ㗔ၞߢ޽ࠆߎߣ
߇␜ߐࠇߚ‫ߩߎޕ‬㗔ၞߪ⇹᳓ᕈᱷၮࠍ฽߻ᐔမᕈߩ
㜞޿᭴ㅧࠍᒻᚑߒߡ߅ࠅ‫ޔ‬᳖⚿᥏⴫㕙ߦኻߒߡ⋧⵬⊛ߦ⚿วߔࠆߣ੍᷹ߐࠇߚ‫ޔߚ߹ޕ‬቟੗ࠄߪࡢࠞࠨ
ࠡ↱᧪ߩ Ca2+㕖ଐሽဳ AFP ࠍኻ⽎ߣߒߡ‫ޔ‬Ca2+⚿ว࡞࡯ࡊߦ ሽ࿷ߔࠆ Thr ߳ᄌ⇣ࠍዉ౉ߒਇಓᵴᕈ
ߩᄌൻࠍ⸃ᨆߒߚ‫⚿ߩߘޕ‬ᨐ‫ޔ‬Thr ஥㎮ߩ᳓㉄ၮ߇᳖ ⚿᥏߳ߩ⚿วߦ㑐ਈߒߡ޿ࠆߎߣࠍႎ๔ߒߚ 17)‫ޕ‬ߎߩߎߣߪ‫ޔ‬I ဳ AFP ߿ Ca2+ 㕖ଐ ሽᕈߩ II ဳ
AFP ߥߤߦ੍ᗐߐࠇߚ AFP ߩ⇹᳓ᕈ㗔ၞ߇᳖⚿᥏ߣߩ⋧੕૞↪ߦߟ޿ߡ㊀ⷐߢ޽ࠆߣ޿߁⍮⷗ߣߪ⇣ߥߞߡ޿ࠆ‫ޔࠄ߆ߣߎߩߎޕ‬AFPII ߪ C ဳ࡟ࠢ࠴ࡦ
ࠍ␲వಽሶߣߒߡ㛽ᩰ᭴ㅧࠍ଻ᜬߒߥ߇ࠄㅴൻߒ‫ޔ‬᳖⚿᥏ߣߩ⋧੕૞↪᭽ᑼߦߟ޿ߡᄙ᭽ᕈࠍ᦭ߒߡ޿
ࠆߣ⠨߃ࠄࠇࠆ‫ޕ‬
 AFPIII ߪർ☨ߩർ᧲ㇱᴪጯߦ↢ᕷߒߡ޿ࠆࠥࡦ
ࠥ⑼ߦዻߔࠆ㝼㘃߆ࠄ⷗಴ߐࠇߚ 10)‫ޕ‬ಽሶ㊂ߪ 6.5
ޯ7kDa ߢ޽ࠅ‫⚵㉄ࡁࡒࠕޔ‬ᚑߦ஍ࠅ߇ߥߊ➅ࠅ㄰ߒ㈩೉߿ಽሶౝ SS ᨞ᯅ߽᦭ߒߥ޿‫ޕ‬AFPIII ߩ․ᓽ
ߩ㧝ߟߪ㧝୘ߩ㝼૕ߩਛߦᢙᄙߊߩ AFPIII ࠕࠗ࠰ࡈࠜ࡯ࡓ߇⊒⃻ߐࠇߡ޿ࠆߎߣߢ޽ࠆ‫ޕ‬ฦࠕࠗ࠰ࡈࠜ࡯ࡓ ߪਛᕈ pH 㗔ၞߦ߅ߌࠆࠗࠝࡦ੤឵᮸⢽߳ߩ⚿วߩ઀ᣇߦࠃߞߡ‫ޔ‬㒶ࠗࠝࡦ੤឵᮸⢽ߢ޽ࠆ
QAE-Sephadex ߦ⚿วߔࠆ QAE ࠲ࠗࡊ(Ⴎၮᕈ஥ߩ pI ࠍ ᦭ ߔ ࠆ ) ‫ ޔ‬㓁 ࠗ ࠝ ࡦ ੤ ឵ ᮸ ⢽ ߢ ޽ ࠆ
SP-Sephadex ߦ⚿วߔࠆ SP ࠲ࠗࡊ(㉄ᕈ஥ߩ pIࠍ᦭ߔࠆ)ߦᄢ೎ߐࠇࠆ‫ޕ‬ർᶏ㆏᧲ㇱᴪጯ᳓ၞߢᄢ㊂ߦ᝝₪ߐࠇࠆࠥࡦࠥ⑼㝼㘃߆ࠄᓧࠄࠇࠆ AFPIII ߩ႐ว‫ߩࡓ࡯ࠜࡈ࠰ࠗࠕޔ‬ᢙߪ 13 ߦ߽਄ࠅ‫ࠇߙࠇߘޔ‬ߩਇಓᵴᕈߪᄢ߈ߊ⇣ߥࠆ 12)‫ޕ‬AFPIII ߪ 3 ᧄߩ⍴޿Ǫ-ࠬ࠻࡜ࡦ࠼ߦࠃߞߡᒻᚑߐࠇࠆੑ⚵ߩㅒᐔⴕǪࠪ࡯࠻ࠍ㛽ᩰߦ߽ߟ⃿⁁Ⱞ⊕⾰ߢ޽ࠅ‫ߩࠄࠇߎޔ‬Ǫࠪ࡯࠻ߦࠃߞߡ᜽߹ࠇߚಽሶߩਛᔃㇱߪ⇹᳓ᕈࠦࠕࠍᒻᚑߒߡ޿ࠆ‫ޕ‬1 ᧄߩǪ-ࠬ࠻࡜ࡦ࠼ߣߘߩㄭற
ߩ࡞࡯ࡊߦࠃߞߡᒻᚑߐࠇߡ޿ࠆᐔ㕙ᕈߩ㜞޿ಽሶ⴫㕙߇᳖⚿᥏⚿วㇱ૏ߣ੍ᗐߐࠇ‫ޔ‬หㇱ૏ߪⷫ᳓ᕈ
ᱷၮߣ⇹᳓ᕈᱷၮ ߩਔᣇࠍ฽߻‫ࡓ࡯ࠜࡈ࠰ࠗࠕޕ‬㑆ߢਇಓᵴᕈ߇⇣ߥࠆߎߣߪ‫⚿߇ࠇߙࠇߘ ޔ‬วߢ߈ࠆ᳖⚿᥏㕙ߩ⒳㘃ߣ⚿วߩᒝߐ߇㑐ଥߒߡ޿ࠆߣ⠨߃ࠄࠇߡ޿ࠆ‫ޕ‬ ਄⸥ߦട߃ߡ‫ޔ‬1996 ᐕߦߪࠞࠫࠞߩ㠦ߦዪ࿷ߔࠆ
⊹ဳ㧔ࠬࠠࡦ࠲ࠗࡊ㧕AFP ߣ๭߫ࠇࠆ⒳㘃߇ႎ๔ߐ

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ಽሶ߇ሽ࿷ߔࠆߎߣߦߥࠅ‫ ߩߤޔ‬AFP ߇᦭↪߆ߟ㊂↥น⢻ߢ޽ࠆ߆ࠍ⷗ᭂ߼ࠆߎߣߪኈᤃߢߪߥ޿‫ޕ‬ᣂߚߥ㝼㘃 AFP ಽሶ߇੹ᓟ⊒⷗ߐࠇࠆน⢻ᕈ߽޽ࠆ‫ޕ‬ߥ߅‫⪺ޔ‬⠪ࠄߪ 2012 ᐕߦ⩶㘃↱᧪ߩ AFP ߩ㧟ᰴరಽሶ᭴ㅧߩ⸃᣿ߦᚑഞߒߚ㧔࿑㧝㧕9)‫⩶ߩߎޕ‬㘃↱᧪ߩ AFP ߩน⢻ᕈߦߟ޿ߡߪᯏળࠍᡷ߼ߡ⚫੺ߒߚ޿‫ޕ‬ࠇ 4)‫ޔ‬2004 ᐕߦߪᾲ቟ቯᕈߦߪᰳߌࠆ߽ߩߩᭂ߼ߡ
ᒝ޿ਇಓᵴᕈࠍ␜ߔᣂߒ޿࠲ࠗࡊߩ㝼㘃 AFP ߇⷗಴ߐࠇߡ޿ࠆ 11)‫╙ޕ‬㧠ߩ࠲ࠗࡊߦዻߔࠆ AFP ࠍࠞࠫࠞ߇᦭ߒߡ޿ࠆߣߩ⺰ᢥ߽ 1997ޯ8 ᐕߦ㧟ႎߛߌ⊒⴫ߐࠇߚ߇‫ࠍࠅ⺋ߩ⸃⷗ߩߎޔ‬ᜰ៰ߒߚ⺰ᢥ߇ 2008ᐕߦ಴ߐࠇߡ޿ࠆ 3)‫ߦ߁ࠃߩߎޕ‬㝼㘃↱᧪ߩ AFP ߦ㒢ߞߡ߽ⶄᢙߩ࠲ࠗࡊ߇⷗಴ߐࠇߡ߅ࠅ‫ޔ‬ฦ‫ߩޘ‬᳖⚿᥏⚿วᯏ⢻ߩ⸃᣿߇ᓙߚࠇߡ޿ࠆ‫ᤨ⃻ޕ‬ὐߢࠕࠗ࠰ࡈࠜ࡯ࡓ߹ߢࠍ฽߼ࠆߣ⊖એ਄ߩ⇣ߥࠆ㝼㘃 AFP

࿑㧝. 㝼㘃↱᧪ AFP㧔AFPIޯIII㧕߅ࠃ߮⩶㘃↱᧪ AFP㧔FingalAFP㧕ߩਥ㎮㛽ᩰ᭴ㅧߩ࡝ࡏࡦࡕ࠺࡞‫ߕ޿ޕ‬ࠇߩ AFP ߽᳖⚿᥏㕙ߦኻߒߡ․⇣⊛ߦ⚿วߢ߈ࠆಽሶ⴫㕙ࠍ᦭ߒߡ޿ࠆ‫ޕ‬ࠍᷝടߒߚ⦡᳓ࠍಓ⚿ߐߖߚ႐วߦߪ‫ޔ‬AFP Ớᐲࠍ
0.1 ߆ࠄ 0.5mg/ml ߹ߢჇടߐߖࠆߦᓥ޿⊕ߞ߸ߐ߇ᶖ߃‫ޔ‬᳖ߩ⋓ࠅ਄߇ࠅ߽ᛥ߃ࠄࠇࠆߎߣ߇ಽ߆ࠆ‫ޕ‬ߔߥࠊߜ AFP ࠍᷝടߒߚ⦡᳓ߢߪో૕߇㞲߿߆ߦ⌕⦡ߐࠇߚ߹߹ಓ⚿ߔࠆ‫ޔߪࠇߎޕ‬AFP ߇୘‫࠷ߩޘ‬ࡉ᳖ߦ⚿วߒߘࠇࠄߩ⴫㕙ࠍⷒ޿‫ࠄࠇߘޔ‬ห჻ߩⲢวࠍ㒖ᱛߔࠆ⚿ᨐߣߒߡ‫⦡ޔ‬ᚑಽࠍឃ㒰ߔࠆജ߿૕
Ⓧ⤘ᒛࠍᛥ߃ࠆὑߣផኤߐࠇࠆ‫ ߥ߁ࠃߩߎޕ‬AFP ߩ௛ ߈ ߪ ‫ ޟ‬ౣ ⚿ ᥏ ൻ 㒖 ኂ ᯏ ⢻ (Recrystallization
Inhibition)‫ߣޠ‬๭߫ࠇࠆߎߣ߽޽ࠆ‫ޕ‬᣸⯻‫ޔ‬ᬀ‛‫⩶ޔ‬㘃↱᧪ߩ AFP ߦߟ޿ߡ߽ᵴᕈߩᒝߐߦ㆑޿ߪ޽ࠆ
ߌࠇߤ߽᳖ߩౣ⚿᥏ൻ㒖ኂᯏ⢻߇⹺߼ࠄࠇߡ޿ࠆ‫ޕ‬ߒ߆ߒ‫ޔ‬᳖⚿᥏ࠍ․೎ߥ౐ᣇਔ㍝ဳ߿㊎⁁⚿᥏ߦ߹ߢᚑᒻߒߚ⁁ᘒߢౣ⚿᥏㒖ኂᯏ⢻ࠍ⊒ើߢ߈ࠆߩߪ‫ޔ‬੹ߩߣߎࠈ㝼㘃 AFP ߛߌߢ޽ࠆ‫ ߦ߁ࠃߩߎޕ‬AFP
ߪಓ⚿⁁ᘒߦ޽ࠆ฽᳓‛ߩౝㇱࠍ࠷ࡉ᳖ߢḩߚߔ௛㝼㘃↱᧪ AFP ߩᯏ⢻ 㝼㘃↱᧪ߩ AFP ࠍ↪޿ߚಓ⚿ᛛⴚߩℂ⸃ߦᓎ┙ߟ◲නߥታ㛎଀ࠍ࿑㧞ߦ␜ߔ‫ߕ߹ޕ‬㧠ߟߩ⚕ࠦ࠶ࡊߦ⦡᳓ࠍ 50ml ߕߟ↪ᗧߔࠆ‫ޕ‬ᰴ޿ߢ‫ޔ‬ฦ‫ߦޘ‬ኻߒߡ㝼૕߆ࠄ♖⵾ߒߚ AFP ߩ☳ᧃࠍ 0‫ޔ‬0.1‫ ޔ‬0.2‫ޔ‬
0.5mg/ml ߦߥࠆࠃ߁ߦട߃ࠆ‫ࠍࠄࠇߎޕ‬᳢↪ߩ಄ಓᐶ㧔⚂-20͠㧕ߦ౉ࠇߡಓࠄߖߚ⚿ᨐ߇࿑㧞ߩ᭽ߦߥࠆ‫ޔߜࠊߥߔޕ‬AFP ࠍട߃ߡ޿ߥ޿᳖㧔Ꮐ┵ߩࠦ࠶ࡊ㧕ߢߪಓߞߚ᳓߇⊕ߊߥࠅ᳖ߩਛᄩㇱ߇⊕ߊ⋓ࠅ਄߇ࠆ‫ޕ‬᳖ߪήᢙߩ࠷ࡉ᳖㧔᳖ᩭ㧕߇Ⲣวߒߚ߽ߩ
ߢ޽ࠆ߇‫ޔ‬᳓ಽሶએᄖߩ‛⾰ߪߎߩⲢวߦෳടߢ߈
ߥ޿‫⚿ߩߘޕ‬ᨐ‫⦡ޔ‬ᚑಽߪⲢวߔࠆ࠷ࡉ᳖߆ࠄ⥄േ⊛ߦឃ㒰ߐࠇࠆ‫ޕ‬ឃ㒰ߐࠇߚ⦡ᚑಽߪᧂಓ⚿ߩㇱಽߔߥࠊߜ⚕ࠦ࠶ࡊߩਛᄩઃㄭߦỚ❗ߐࠇࠆ‫ߒ߁ߎޕ‬ߡ⦡߇ᛮߌߚ᳖߇⊕ߊ⷗߃ࠆ‫ޔߚ߹ޕ‬᳓߇᳖ߦᄌൻ
ߔࠆ㓙ߩ૕Ⓧ⤘ᒛ߇ਛᄩㇱࠍ⋓ࠅ਄ߍࠆ‫৻ޕ‬ᣇ‫ޔ‬AFP

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ᢱ‫ޔ‬㜞ಽሶࠥ࡞╬ߩౝㇱࠍ࠷ࡉ᳖ߢḩߚߔᣂߒ޿ಓ
⚿଻ሽᛛⴚࠍ㐿⊒ߢ߈ࠆߣ⠨߃ࠄࠇࠆ 13)‫ޕ‬߈ࠍߔࠆ‫ࠍࠇߎޕ‬ᔕ↪ߔࠆߎߣߢ‫ޔ‬ടᎿ㘩ຠ‫࡯ࠬޔ‬ࡊ㘃‫ޔ‬᳖⩻ሶ㘃‫ࠎ߼ޔ‬㘃‫ࡦࡄޔ‬㘃‫ޔ‬㊁⩿‫ޔ‬ᨐታ‫ޔ‬⒳ሶ‫ޔ‬ᷡᶭ㘶ᢱ᳓‫ޔ‬㈬㘃‫ޔ‬ක≮ຠ‫ޔ‬ൻ♆ຠ‫ࠢࡦࠗޔ‬㗻

࿑㧞㧚AFP ߩಓ⚿Ớ❗ᛥ೙ലᨐࠍ␜ߔታ㛎ߩ଀‫ޕ‬਄Ბ㧦 AFPIII ߩỚᐲ߇ 0‫ޔ‬0.1‫ޔ‬0.2‫ޔ‬0.5mg/ml ߦߥࠆࠃ߁
ߦ⺞⵾ߒߚ⌕⦡᳓ࠍг18͠ߢ৻᥅ಓ⚿ߐߖߚᓟߩ᳖ߩ↹௝‫ޕ‬ਅᲑ㧦ㅢᏱߩ᳓ṁᶧࠍಓࠄߖߚߣ߈ߦ⿠ߎࠆṁ⾰ߩಓ⚿Ớ❗⃻⽎ߩᮨᑼ࿑㧔Ꮐ㧕‫ޕ‬AFP ࠍᷝടߔࠆߣ᳖ߩ࠷ࡉߣṁ⾰ᚑಽߩᷙว⁁ᘒߣߥࠅಓ⚿Ớ❗⃻⽎߇ᛥ೙ߐࠇࠆ‫ޕ‬ 㝼㘃↱᧪ AFP ߩ⚦⢩଻⼔ᯏ⢻ࠍ⋥ᗵ⊛ߦℂ⸃ߔ
ࠆߚ߼ߩታ㛎⚿ᨐࠍ࿑㧟ߦ␜ߔ‫౒ߪࠄࠇߎޕ‬ὶὐ࡟࡯ࠩ࡯㗼ᓸ㏜ࠍ↪޿ߡ᠟ᓇߒߚ AFP ߹ߚߪ࠙ࠪⴊ
ᷡࠕ࡞ࡉࡒࡦ㧔BSA㧕ሽ࿷ਅߦ߅ߌࠆ㧝୘ߩࡅ࠻⢄⤳↱᧪⚦⢩㧔HepG2㧕ߩ↹௝ߢ޽ࠆ‫⹜ޕ‬ᢱ᷷ᐲߪ 30͠ߢ޽ࠅ‫ޔ‬㕖ಓ⚿⁁ᘒߢߩⷰኤ⚿ᨐߢ޽ࠆߎߣߦᵈᗧߐࠇߚ޿‫ޕ‬AFP ߣ BSA ߦߪߘࠇߙࠇⰯశ‛⾰ࠍ⚿วߐߖߡ޿ࠆ‫ޕ‬࿑㧟A ߪ AFP ሽ࿷ਅߢߩห㗼ᓸ㏜ࠬ࡜
ࠗࠬ↹௝ߢ޽ࠅ‫⚦ޔ‬⢩⴫㕙ߩ⢽⾰ੑ㊀⤑ઃㄭ߇߹ࠎ
ߴࠎߥߊశߞߡ޿ࠆߎߣ߇್೎ߢ߈ࠆ‫ޕ‬૏⟎ࠍᄌ߃ߡ᠟ᓇߒߚ 16 ᨎߩࠬ࡜ࠗࠬ↹௝߆ࠄวᚑߒߚ⚦⢩ో૕ߩࠗࡔ࡯ࠫ㧔࿑㧟㨏㧕ߦ߅޿ߡ߽ AFP ߩ⚦⢩⤑߳ߩๆ⌕߇⏕⹺ߢ߈ࠆ‫৻ޕ‬ᣇ‫ޔ‬ห᧦ઙߢ᠟ᓇߒߚ
BSA ሽ࿷ਅߦ߅ߌࠆ HepG2 ߩ↹௝ߢߪ‫ޔ‬BSA ߽޿ߊ߱ࠎ⢽⾰ੑ㊀⤑ઃㄭߦๆ⌕ߒߡߪ޿ࠆ߽ߩߩ‫ߘޔ‬ߩኒᐲߪ AFP ߩߘࠇߦᲧߴߡ⪺ߒߊૐ޿‫ߜࠊߥߔޕ‬㝼㘃↱᧪ߩ AFP ߪࡑࠗ࠽ࠬߩ᷷ᐲၞߢߪ᳖⚿᥏ߦ⚿วߒ‫ߩࠬ࡜ࡊޔ‬᷷ᐲၞߦ߅޿ߡߪ⚦⢩⤑ߦ⚿วߔࠆᯏ⢻ࠍ᦭ߒߡ޿ࠆ‫⪺ޕ‬⠪ࠄߪ‫ ࠬ࡜ࡊޔ‬4͠ߩ㕖ಓ⚿᷷ᐲၞߦ߅޿ߡ‫ޔ‬AFP ߇ HepG2 ⚦⢩ߩ↢ሽ₸ࠍ㘧べ⊛ߦ㜞߼ࠆߎߣࠍ᣿ࠄ߆ߦߒߚ 7)‫ޔߦᦝޕ‬㔚᳇ൻቇ⊛ታ㛎ߩ⚿ᨐࠍၮߦߒߡ HepG2 ⚦⢩ߪᤨ㑆⚻ㆊߣߣ߽ߦ⤘ᒛߒߡ⎕ⵚᱫߦ⥋ࠆ߇‫ޔ‬AFP ߩ⚦⢩⴫㕙ๆ⌕߇ߘߩ⤘ᒛࠍᛥ೙ߔࠆߎߣߦࠃߞߡ HepG2 ߩ⎕ⵚᱫࠍㆃࠄߖࠆߩߢߪߥ޿߆㧫ߣផኤߒߚ 8)‫ߎޕ‬ߩ⚦⢩଻⼔ലᨐߪච⒳㘃એ਄ߩ⚦⢩ߦߟ޿ߡ⏕⹺ߐࠇߡ߅ࠅ‫ޔ‬ᣂߒ޿⚦⢩⤑଻⼔೷ߣߒߡߩ AFP ߩᵴ↪
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)࿑㧟㧚 AFP ߹ߚߪ࠙ࠪⴊᷡࠕ࡞ࡉࡒࡦ㧔BSA㧕ሽ࿷ਅߦ߅ߌࠆࡅ࠻⢄⤳↱᧪⚦⢩㧔HepG2㧕ߩ౒ὶὐ࡟࡯ࠩ
࡯㗼ᓸ㏜↹௝‫ޕ‬ฦ࠲ࡦࡄࠢ⾰ߦߪⰯశ࡜ࡌ࡞ࠍᣉߒߡ޿ࠆ‫ޕ‬਄Ბ㧦㧔㧭㧕Ⱟశ࡜ࡌ࡞ AFP ߩ⚦⢩⤑ๆ⌕ࠍ␜ߔ㗼ᓸ㏜ࠬ࡜ࠗࠬ↹௝‫ޔ‬㧔㧯㧕หࠬ࡜ࠗࠬ↹௝ࠍ㊀ߨߡวᚑߒߚ⚦⢩ో૕ࠍ⴫ߔ↹௝‫ޕ‬ਅᲑ㧦㧔㧮‫ޔ‬㧰㧕Ⱟశ
࡜ࡌ࡞ BSA ߇ HepG2 ⚦⢩ߦ⚿วߔࠆ᭽ሶࠍ⴫ߔ↹௝‫ޕ‬╬ߦᷝടߒߚ႐วߦߘߩ㘩ᗵ߿ຠ⾰߇ᡷༀߐࠇࠆߎ
ߣ߇ಽ߆ߞߡ޿ࠆ‫ޕ‬ 㝼㘃↱᧪ AFP ߪ‫ޔ‬એਅߩఝࠇߚ․ᓽࠍ஻߃ߡ޿
ࠆ㧦㧝㧕☻♖⵾ຠߣߒߡ߽㜞⚐ᐲຠߣߒߡ߽↢↥น⢻㧔ᄤὼ‛㧕
‫ޔ‬㧞㧕᳖⚿᥏ࠍ౐ᣇਔ㍝ဳ߿㊎⁁ߥߤߩ․ቯߩᒻ⁁ߦᚑᒻߢ߈ࠆ㧔ᵈ㧚AFPIޯIII ߅ࠃ߮
AFGP㧕‫ޔ‬㧟㧕ઁߩ AFP ࠃࠅ߽ಽሶࠨࠗ࠭߇㕖Ᏹߦዊߐ޿㧔ᵈ㧚AFPI ߣ AFPIII㧕‫ޔ‬㧠㧕᳓ṁᕈ߇ᭂ߼ߡ㜞޿㧔ᵈ㧚AFPI ߣ AFGP㧕‫ޔ‬㧡㧕᳓ߦṁ⸃ߒߚᓟߩ⠴ᾲᕈߣ⠴ਭᕈߦఝࠇߡ޿ࠆ㧔ᵈ㧚AFPI ߣ AFGP㧕‫ޔ‬㧢㧕⚦⢩଻⼔ലᨐࠍ⊒ើߔࠆ㧔ᵈ㧚AFPI‫ޔ‬AFPIII ߅ࠃ߮ AFGP㧕‫ޕ‬ᓥߞߡ‫ޔ‬㝼㘃 AFP ߪࠕࠗࠬࠢ࡝࡯ࡓ
߿ಓ⚿߼ࠎ㘃ߩߺߥࠄߕ‫ޔ‬කቇ⊛ಽ㊁ࠍ฽߻ࠃࠅᐢ
▸߆ߟ♖ኒߥ↪ㅜߦኻߒߡ߽ᔕ↪߇น⢻ߢ޽ࠆ‫ޕ‬ᶧ૕⓸⚛߿․೎ߥⵝ⟎ࠍᔅⷐߣߒߥ޿⋭ࠛࡀ࡞ࠡ࡯ᕈ
ࠍ᦭ߒߡ޿ࠆߎߣ߇․ߦ㝼㘃 AFP ᛛⴚ߇᦭ߔࠆᄢ
߈ߥ․ᓽߢ޽ࠆ‫ޕ‬ᣣᧄߪ਎⇇╙㧢㧝૏ߩ࿖࿯㕙Ⓧࠍ
ᠩߔዊߐߥ࿖ߛ߇‫ޔ‬ឃઁ⊛⚻ᷣ᳓ၞ㧔㗔ᶏࠍ฽߻㧕ߩᐢߐߢߪ਎⇇╙㧢૏ߦߥࠆ‫ޕ‬㝼㘃 AFP ߪߎߩᐢᄢ
ߥ᳓ၞߢ᝝₪ߐࠇࠆ㝼㘃߆ࠄήዧ⬿ߦᄢ㊂↢↥ߔࠆߎߣ߇ේℂ⊛ߦߪน⢻ߣ⸒߃ࠆ‫੹ޕ‬ᓟ߽ၮ␆ߣᔕ↪

㝼㘃↱᧪ AFP ߩᵴ↪ᛛⴚߣ੹ᓟߩዷᦸ
 AFP ߩ᳖⚿᥏⚿วᯏ⢻ߪ౒㔀‛ߩሽ࿷ਅߢ߽චಽ
ߦ⊒ើߐࠇࠆ‫♧ޕ‬㘃‫ޔ‬Ⴎ‫ޔ࡞࡯ࡠ࠮࡝ࠣޔ‬ዊಽሶ㊂ࡍࡊ࠴࠼ߥߤߪ AFP ߩਇಓᯏ⢻ࠍ㜞߼ࠆ௛߈ࠍߔ
ࠆ‫ޕ‬AFPI ߪࠛ࠲ࡁ࡯࡞ሽ࿷ਅߢ߽᳖⚿᥏⚿วᯏ⢻ࠍᄬࠊߥ޿੐߇੍஻⊛ታ㛎߆ࠄ␜ߐࠇߡ޿ࠆ‫߽ߣ߽ޕ‬ߣ AFP ߇฽߹ࠇߡ޿ࠆേᬀ‛ߩ૕ᶧਛߦߪᢙᄙߊ
ߩ‛⾰߇ᷙ࿷ߒߡ߅ࠅ‫ޔ‬AFP ߪ౒㔀‛ߩᓇ㗀ࠍฃߌߦߊ޿ࠃ߁ߦ಴᧪ߡ޿ࠆߩ߆߽⍮ࠇߥ޿‫ޔߜࠊߥߔޕ‬ᛛⴚᔕ↪ߦߣߞߡᔅߕߒ߽ AFP ߩ㜞⚐ᐲຠߪᔅⷐ
ߢߪߥ޿‫߫߃଀ޕ‬಄⫾ᾲ߿㘩ຠߥߤߩಽ㊁ߢߪ AFPߩᄤὼ‛޽ࠆ޿ߪㆮવሶ⚵឵߃‛ߩ☻♖⵾ຠࠍ૶߁
ߎߣ߇ߢ߈ࠆ‫ޕ‬ታ㓙ߦ‫ޔ‬࡙࠾࡝ࡃ࡯␠ߪ․ቯߩ AFPIIIࠕࠗ࠰ࡈࠜ࡯ࡓߩㆮવሶ⚵឵߃‛ߩ☻♖⵾ຠࠍᷝട
ߒߚࠕࠗࠬࠢ࡝࡯ࡓࠍ⽼ᄁߒߡ޿ࠆ‫ ߜࠊߥߔޕ‬AFP߇߽ߟ᳖ߩౣ⚿᥏ൻ㒖ኂᯏ⢻ࠍ೑↪ߔࠆߎߣߢࠕࠗ
ࠬࠢ࡝࡯ࡓਛߦ↢ᚑߒߚ᳖⚿᥏ߩ☸ᓘࠍዊߐߊ଻ߚ
ߖࠆߎߣߦᚑഞߒߡ޿ࠆ‫ޕ‬ᓥ᧪‫ࡓ࡯࡝ࠢࠬࠗࠕޔ‬ౝߩ᳖ߩ☸ᓘߪᄙ㊂ߩ੃⢽⢌ಽࠍട߃ࠆߎߣߢዊߐߊ
ߒߡ߈ߚ߇‫ޔ‬ઍࠊࠅߦ AFP ࠍട߃ࠆߎߣߢ੃⢽⢌ಽࠍᷫࠄߔߎߣ߇಴᧪ࠆߣߐࠇࠆ‫ޔߚ߹ޕ‬ಓ⚿߼ࠎ㘃

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ߩਔ㕙߆ࠄ㝼㘃 AFP ߩ⎇ⓥ㐿⊒ࠍ᛼ߒㅴ߼ᣣᧄߩ
․ᓽࠍ↢߆ߔࠃ߁ߥᣂᛛⴚࠍഃߞߡⴕ߈ߚ޿‫ޕ‬

9)

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Journal of Japanese Society for Extremophiles (2012) Vol.11, 70-74

Fujisawa M.a,b

Isolation of alkaliphiles, which can grow by oxidative phosphorylation underhigh lithium ion concentration instead of sodium ion and its physiology,
predicted from comparative genome analysis with sodium dependent ones.
㜞Ớᐲߩ࡝࠴࠙ࡓࠗࠝࡦሽ࿷ਅߢ㉄ൻ⊛࡝ࡦ㉄ൻߦࠃߞߡ↢⢒ߔࠆᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩන㔌ߣࠥࡁࡓ߆ࠄ੍᷹ߐࠇࠆ↢ℂ⊛ᕈ⾰᧲ᵗᄢቇ ↢๮⑼ቇㇱ 㘩ⅣႺ⑼ቇ⑼᧲ᵗᄢቇࡃࠗࠝ࡮࠽ࡁࠛ࡟ࠢ࠻ࡠ࠾ࠢࠬ⎇ⓥ࠮ࡦ࠲࡯⮮Ỉ ⺈aFaculty of Food Life Sciences, Toyo University, 1-1-1 Izumino, Itakura-machi, Oura-gun, Gunma, 374-0193, JAPAN
Bio-Nano Electronics Research Center, 2100 Kujirai, Kawagoe, Saitama, 350-8585, JAPANb

Correspondence author: Makoto Fujisawa, m_fujisawa@toyo.jp
Tel: +81-276-82-9025 Fax: +81-276-82-9025
Received: October 27, 2012 / Accepted: November 6, 2012ߪߓ߼ߦ pH 9.0 એ਄ߣ޿߁㜞޿ࠕ࡞ࠞ࡝ⅣႺਅߢ⥋ㆡ↢⢒ࠍ␜ߔᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߪ‫࡞ࠕߦ߁ࠃߩߤߪ‛↢ޔ‬
ࠞ࡝ⅣႺߦㆡᔕߔࠆߩ߆‫↢߇‛↢ߚ߹ޔ‬ሽߒ߁ࠆ pHߩ਄㒢ߪߤߩ⒟ᐲߥߩ߆ߣ޿ߞߚᩮᧄ⊛ߥ໧㗴߿‫ޔ‬ࠕ࡞ࠞ࡝ⅣႺਅߢᯏ⢻⊛ߦ௛ߊ㉂⚛ࠍ↥↢ߔࠆߣ޿߁ὐߢ⥝๧ᷓ޿⎇ⓥኻ⽎ߢ޽ࠆ‫ޕ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩ․ᓽߩ৻ߟߣߒߡ‫(ࡦ࠻ࡠࡊޔ‬H+)ߩዋߥ޿ⅣႺਅߢ↢⢒ߔࠆߚ߼ߦ࠽࠻࡝࠙ࡓࠗࠝࡦ(Na+)ࠍ೑↪ߔࠆߎߣ߇᜼ߍࠄࠇࠆ 1, 6, 9)‫ޔߦ⥸৻ޕ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߪ⚦⢩ౝ pH ࠍਛᕈઃㄭߦ଻ߞߡ߅ࠅ‫⚦߇ࠇߎޔ‬⢩ౝᄖ߳ߩ Na+ߩᓴⅣ♽ߣ౒ᓎߒߡ޿ࠆ‫ޔࠅ߹ߟޕ‬Na ࠴ࡖࡀ࡞‫ޔ‬Na+㚟േဳߴࠎᲫࡕ࡯࠲࡯‫ޔ‬Na+/ࠕࡒࡁ㉄౒ャㅍ૕ߥߤߦࠃߞߡ⚦⢩ౝߦࠃࠅขࠅㄟ߹ࠇ
ߚ Na+߇‫ޔ‬Na+/H+ࠕࡦ࠴ࡐ࡯࠲࡯ߦࠃߞߡឃ಴ߐࠇ
ࠆ㓙ߦ H+߇ขࠅㄟ߹ࠇ‫⚦ޔ‬⢩ౝ߇㉄ᕈൻߐࠇࠆ‫৻ޕ‬ᣇ‫ޔ‬Bacillus foraminis CV53T ߩࠃ߁ߦ↢⢒ߦ Na+ࠍᔅⷐߣߒߥ޿߽ߩ߽⷗ߟ߆ߞߡ߅ࠅ 10)‫ߥ߁ࠃߩߎޔ‬↢‛ߢߪ‫(ࡦࠝࠗࡓ࠙࡝ࠞߊࠄߘ߅ޔ‬K+)ߥߤ೎ߩࠗ
ࠝࡦࠍ೑↪ߒߚ⚦⢩ౝ㉄ᕈൻᯏ᭴߇ሽ࿷ߔࠆߣ⠨߃ࠄࠇߡ޿ࠆ 9)‫ޕ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩߎߩ․ᓽ߆ࠄ⠨߃ࠆߣ‫ޔ‬Na+߿ K+એᄖߩ᭽‫ߥޘ‬㓁ࠗࠝࡦࠍᏁߺߦ೑
↪ߒ‫⚦ޔ‬⢩ౝࠍ㉄ᕈൻߔࠆߎߣߩߢ߈ࠆᅢࠕ࡞ࠞ࡝
ᕈ⚦⩶߇ሽ࿷ߒߡ޿ߡ߽߅߆ߒߊߪߥ޿‫ޕ‬ᚒ‫ޔߪޘ‬ߎߩᕈ⾰ࠍᵴ↪ߔࠆߎߣߢ‫ޔ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩᣂ

Abstract Alkaliphilic bacterial organisms have an
optimal growth pH above 9.06). They can maintain thecytoplasmic pH close to neutral pH under such high pH
condition. In general, sodium transport cycles arecoupled to the cytoplasm acidification. Bacillus
pseudofirmus OF4, an exclusively studied facultative
alkaliphile whose genome has been sequenced8), cangrow on malate by oxidative phosphorylation and it
requires sodium ion at more than 10 mM at pH 10.59).
However, some of alkaliphiles such as Bacillus foraminis
CV53T are assumed to be independent on sodium ion10).
In addition, contribution of intracellular acid production
to it has been suggested from the fact that B.
pseudofirmus OF4 could maintain the intracellular pH at
9.2 on glucose without sodium when the cells wereupshifted from pH 8.5 to 10.59). Exploring alkaliphilic
bacteria, which grows by oxidative phosphorylation andrequires much lower sodium would be one of thechallenges to find a novel alkaline environment
adaptation.
Keywords: alkaliphiles, Bacillus, genome, ion
transporters, cell surfaceࠠ࡯ࡢ࡯࠼: ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶‫ޔࡓࡁࠥޔࠬ࡞࠴ࡃޔ‬ࠗࠝࡦャㅍ૕‫⚦ޔ‬⢩⴫ጀ
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)↢⢒ߦ 10 mM ⒟ᐲߩ Na+ࠍᔅⷐߣߔࠆ 7)‫ߥߒ߆ߒޕ‬߇ࠄ‫ޔ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩ Li+ߩⷐ᳞ᕈߦߟ޿ߡߪ‫ޔ‬߹ߞߚߊ⺞ߴࠄࠇߡ޿ߥ߆ߞߚ‫ޕ‬ ߘߎߢᚒ‫ ߦ߼ߓߪޔߪޘ‬B. pseudofirmus OF4 ᩣߩ↢⢒ߦ߅ߌࠆ Li+ߩᓇ㗀ࠍ⺞ߴߚ‫ⴣ✭ޕ‬೷ߣߒߡ὇㉄࠽࠻࡝࠙ࡓ‫࡯ࠡ࡞ࡀࠛޔ‬Ḯߣߒߡ࡝ࡦࠧ㉄࠽࠻࡝࠙ࡓࠍ฽߻ඨวᚑၭ࿾(MYE ၭ࿾)߇ B. pseudofirmus
OF4 ᩣߩၭ㙃ߦࠃߊ↪޿ࠄࠇߡ޿ࠆ‫ ߩߎޕ‬MYE ၭ࿾ਛߩ὇㉄࠽࠻࡝࠙ࡓߣ࡝ࡦࠧ㉄࠽࠻࡝࠙ࡓࠍ‫ޔ‬὇㉄ࠞ࡝࠙ࡓߣ࡝ࡦࠧ㉄ࠞ࡝࠙ࡓࠍ↪޿ߡ⺞⵾ߔࠆߎߣߢૐ Na+Ớᐲߩၭ࿾ࠍ૞ᚑߒߚ(KMYE ၭ࿾)‫ߎޕ‬ߩၭ࿾ߦ᭽‫ߥޘ‬Ớᐲߩ Na+߹ߚߪ Li+ࠍᷝടߒ‫ޔ‬B.
pseudofirmus OF4 ᩣߩ↢⢒ታ㛎ࠍⴕߞߚ‫⚿ߩߘޕ‬ᨐ‫ޔ‬
B. pseudofirmus OF4 ᩣߪ pH 10.5 ߦ߅޿ߡߪ 10 mM
એ਄ߩ NaCl ࠍⷐ᳞ߔࠆ߇‫ޔ‬300 mM ߹ߢ LiCl ࠍᷝടߒߡ߽↢⢒ߪߒߥ޿ߎߣ߇᣿ࠄ߆ߣߥߞߚ(࿑ 1ࠍෳᾖ)‫ޕ‬B. pseudofirmus OF4 ᩣߪ pH 7.5 ߩߣ߈‫ޔ‬pH
10.5 ߢⷐ᳞ߔࠆ Na+Ớᐲࠃࠅ߽㜞޿Ớᐲࠍⷐ᳞ߔࠆ
ߎߣ߇⍮ࠄࠇߡ޿ࠆ 7)‫ޔߪࠇߎޕ‬Na+/ࠕࡒࡁ㉄౒ャㅍ૕ߥߤߩ Na+㚟േജࠍ೑↪ߔࠆ‛⾰ャㅍ♽߇ H+ߦࠃࠆ┹ว㒖ኂࠍฃߌࠆߚ߼ߢ޽ࠆߣផኤߐࠇߡ޿ࠆ
9)‫ޔߢߎߘޕ‬B. pseudofirmus OF4 ᩣߩ↢⢒ߦ߅ߌࠆ
LiCl ߩലᨐࠍ pH 8.5‫ޔ‬9.0‫ޔ‬9.5‫ޔ‬10.0‫ޔ‬10.5 ߢ⺞ߴࠆߣ‫ޔ‬pH 10.0 એਅߢ↢⢒߇ⷰኤߐࠇߚ(࿑ 2)‫ޕ‬LiCl Ớᐲ߇㜞ߊߥࠆ߶ߤ⥋ㆡ↢⢒ߪૐ޿ pH ߢⷰኤߐࠇ‫ޔ‬
200 mM LiCl ߢߪ pH 9.0 (࿑ 2A)‫ޔ‬50 mM LiCl ߢߪ
pH 9.5 (࿑ 2B)‫ޔ‬10 mM LiCl ߢߪ pH 9.5-10.0(࿑ 2C)
ߢ⥋ㆡ↢⢒ࠍ␜ߒߚ‫ޔߡߞ߇ߚߒޕ‬Na+ଐሽဳߩ B.
pseudofirmus OF4 ᩣߪ‫ޔ‬ᩕ㙃⚛ߩขࠅㄟߺߥߤߩㆊ⒟ߦ߅޿ߡߪ Na+ߩઍࠊࠅߦ Li+ࠍ೑↪ߢ߈ࠆ߇‫ࠃޔ‬ࠅ㜞޿ pH ߢ⚦⢩ౝࠍ㉄ᕈൻߔࠆ㓙ߦߪ Li+ࠍ߶ߣࠎߤ೑↪ߢ߈ߥ޿߆‫ ߪߚ߹ޔ‬Li+ߩᲥᕈߦኻߔࠆ⠴ᕈᯏ᭴߇⣀ᒙߢ޽ࠆߣ⠨߃ࠄࠇߚ‫ޕ‬ⷙࠕ࡞ࠞ࡝ⅣႺㆡᔕᯏ᭴ߩ⊒⷗࡮⸃᣿ߣᣂߚߥᔕ↪
ಽ㊁ߩ㐿⊒ࠍ⋡ᜰߐߒߡ޿ࠆ‫ౕޕ‬૕⊛ߦߪ‫ޔ‬Na+ߩઍࠊࠅߦ࡟ࠕࡔ࠲࡞ߢ޽ࠆ࡝࠴࠙ࡓࠗࠝࡦ(Li+)ࠍട
߃ߚၭ࿾ࠍ૞ᚑߒ‫ޔ‬Na+ߦଐሽߒߥ޿‫ߪߚ߹ޔ‬ଐሽᕈ߇ૐ޿ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩන㔌ࠍ⹜ߺߚ‫ޔߡߒߘޕ‬ಽ㔌ᩣਛߩ 1 ⩶ᩣߦߟ޿ߡߪ‫⸃ࠍࡓࡁࠥ࠻ࡈ࡜࠼ޔ‬⺒ߒ‫ޔ‬ᣢሽߩ Na+ଐሽဳߩᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩࠥࡁࡓߣᲧセࠍⴕ߁ߎߣߢ‫․ߩߘޔ‬ᓽࠍ᛽಴ߒߚ‫ࠇߎޕ‬ߦࠃࠅ‫ޔ‬Na+㕖ଐሽဳ߹ߚߪૐଐሽဳߩࠕ࡞ࠞ࡝Ⅳ
Ⴚㆡᔕᯏ᭴ߩࡕ࠺࡞ߩ᭴▽ࠍ⋡ᜰߒߚ‫৻ޕ‬ᣇߢ‫ޔ‬Li+ߩ⚿ว࠲ࡦࡄࠢ⾰߇⷗ߟ߆ࠇ߫‫ࠆ޽ߢ࡞࠲ࡔࠕ࡟ޔ‬࡝࠴࠙ࡓߩ࿁෼ᛛⴚߦ೑↪ߢ߈ࠆߒ‫ࡦࠝࠗޔ‬ャㅍ૕ਛߩࠗࠝࡦ⚿วㇱ૏߿ࠗࠝࡦㆬᛯㇱ૏ߩ⍮⷗߇ᓧࠄࠇࠇ߫‫ߩߤߥ∛࡞ࡀࡖ࠴ࡦࠝࠗޔ‬ᴦ≮ߦᔅⷐߥ⍮⷗ࠍᓧࠆߎߣ߇ᦼᓙߢ߈ࠆ‫ޕ‬Na+ଐሽᕈߩᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩ↢⢒ߦ߅ߌࠆ Li+ߩᓇ㗀 Bacillus pseudofirmus OF4 ᩣߪᅢ᳇ᕈߩㅢᕈᅢࠕ
࡞ࠞ࡝ᕈ⚦⩶ߢ‫ޔ‬ਛᕈઃㄭߢ߽⦟ᅢߥ↢⢒ࠍ␜ߔߚ߼‫ߥ⊛⥸৻ޔ‬ਛᕈ⚦⩶ߣߩᲧセ߇ⴕ޿߿ߔ޿ߎߣ߿‫ޔ‬ㆮવሶᎿቇ⊛ᚻᴺࠍ೑↪น⢻ߢ޽ࠆߎߣߥߤ߆ࠄ‫ޔ‬
ࠕ࡞ࠞ࡝ⅣႺㆡᔕᯏ᭴ߩࡕ࠺࡞↢‛ߣߒߡࠃߊ⎇ⓥߐࠇߡ޿ࠆ‫ߥ⊛⥸৻ߪ⩶ᧄޔߚ߹ޕ‬ਛᕈઃㄭߢ↢⢒ߔࠆ⚦⩶ߣห᭽ߦ‫ޔ‬ૐ H+Ớᐲߩࠕ࡞ࠞ࡝ⅣႺਅߦ߅޿ߡ߽ࡊࡠ࠻ࡦ㚟േജࠍ↪޿ߡ㉄ൻ⊛࡝ࡦ㉄ൻࠍⴕߞߡ߅ࠅ‫↢ޔ‬૕ࠛࡀ࡞ࠡ࡯ߩ⎇ⓥಽ㊁ߦ߅޿ߡ߽㊀ⷐߥᓎഀࠍᨐߚߒߡ޿ࠆ 5)‫ߪࡓࡁࠥߩ⩶ᧄޕ‬ᨴ⦡૕
(3.86 Mb)ߣᄢዊ 2 ߟߩࡊ࡜ࠬࡒ࠼(0.29 Mb‫ޔ‬0.10 Mb)
߆ࠄߥࠅ‫ޔ‬2011 ᐕߦోႮၮ㈩೉߇⸃⺒ߐࠇߡ޿ࠆ 8)‫ޕ‬
B. pseudofirmus OF4 ᩣߪ৻⥸⊛ߥᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߣห᭽ߦ‫ޔ‬Na+ߦଐሽߒߚ↢⢒ࠍ␜ߒ‫࡯ࠡ࡞ࡀࠛޔ‬Ḯߣߒߡ࡝ࡦࠧ㉄ࠍ฽ࠎߛ pH 10.5 ߩၭ࿾ߢߪ⥋ㆡ

࿑ 1. B. pseudofirmus OF4 ᩣߣ B. pseudofirmus KU50 ᩣߩ↢⢒ߦ߅ߌࠆ NaCl ߣ LiCl ߩലᨐ᭽‫ߥޘ‬Ớᐲߩ NaCl ߹ߚߪ LiCl ࠍട߃ߚ pH 10.5 ߩ KMYE ၭ࿾ਛߢߩ↢⢒ࠍ⺞ߴߚ‫ޕ‬B. pseudofirmus OF4 ᩣߩ↢⢒ߦ߅ߌࠆ NaCl ߩലᨐ(٨)ߣ LiCl ߩലᨐ(ً)‫ ߮ࠃ߅ޔ‬B. pseudofirmus KU50 ᩣߩ↢⢒ߦ߅ߌࠆ NaCl ߩല
ᨐ(٤)ߣ LiCl ߩലᨐ(ٌ)ࠍߘࠇߙࠇ␜ߒߚ‫ޕ‬71

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)࿑ 2. B. pseudofirmus OF4 ᩣߩ↢⢒ߦ߅ߌࠆ LiCl ߣ pH ߩലᨐ
B. pseudofirmus OF4 ᩣߩ KMYE ၭ࿾ਛߢߩ↢⢒ߦ߅ߌࠆ LiCl ߣ pH ߩലᨐࠍ⺞ߴߚ‫ޕ‬200 mM LiCl ࠍട߃ߚߣ߈ (A)‫ޔ‬50 mM LiCl ࠍട߃ߚߣ߈(B)‫ޔ‬10 mM LiCl ࠍട߃ߚߣ߈(C)ߩ↢⢒ࠍ᭽‫ ߥޘ‬pH ߢ⺞ߴߚ‫࠙࠴࡝ޕ‬ࡓࠍᷝടߒߥ߆ߞߚᤨߩ↢⢒ߪࡄࡀ࡞ A ߩߺߢ␜ߒ‫ޔ‬B ߣ C ߢߪ◲⇛ߩߚ߼⋭⇛ߒߚ‫ޕ‬Na+㕖ଐሽᕈߩᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩන㔌ߣหቯ B. pseudofirmus OF4 ᩣߩ↢⢒ታ㛎ߩ⚿ᨐࠍ߽ߣߦ‫ޔ‬
100 mM એ਄ߩ LiCl ࠍᷝടߒߚ pH 10.5 ߩ KMYE ၭ࿾ࠍᣂⷙᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩಽ㔌ၭ࿾ߣߒߡ↪޿ߚ‫ޕ‬ߎߩ᧦ઙਅߢ↢⢒ߔࠆᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߪ‫ޔ‬ԘNa+߳ߩଐሽᕈ߇ૐ޿‫ޔ‬ԙLi+ߥߤߩ㓁ࠗࠝࡦࠍ⚦⢩ౝߩ㉄ᕈൻߦ೑↪ߢ߈ࠆ‫ޔ‬ԚLi+⠴ᕈ߇㜞޿ߎߣ߇੍ᗐߐ
ࠇߚ‫ޕ‬὇⚛Ḯߦࠣ࡞ࠦ࡯ࠬߢߪߥߊ‫↪ࠍ㉄ࠧࡦ࡝ޔ‬޿ߚℂ↱ߣߒߡߪ‫ޔ‬ህ᳇๭ๆߦࠃࠆ᦭ᯏ㉄ߩ↢ᚑߦࠃߞߡ⚦⢩ౝߩ㉄ᕈൻ߇⿠ߎࠆߣ‫ޔ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩ㓁ࠗࠝࡦ߳ߩଐሽᕈ߇ૐਅߔࠆߎߣ߇⠨߃ࠄࠇߚߚ߼ߢ޽ࠆ 2)‫ޕ‬ᓸ↢‛ߩಽ㔌Ḯߦߪ৻⥸⊛ߥ࿯ფ߿ḓᴧߦട߃‫ ࡝ࠞ࡞ࠕޔ‬pH ࠍ␜ߔ⍹Ἧ⾰ߩ࿯ფ‫ޔ‬࡝࠴࠙ࡓࠍ฽߻࿖ౝᄖߩ࡝ࠪࠕ㔕Უ߿ጤႮߥߤࠍ↪
޿ߚ‫⚿ޕ‬ᨐߣߒߡ‫ ߦߢ߹ࠇߎޔ‬56 ⩶ᩣ߇න㔌ߐࠇߚ‫ޕ‬ߎࠇࠄߩ⚦⩶߆ࠄᨴ⦡૕ DNA ࠍ⺞⵾ߒ‫ޔ‬16S rDNA㗔ၞࠍ PCR ߢჇ᏷ᓟ‫ޔ‬Ⴎၮ㈩೉ࠍ᳿ቯߒ‫⋧ޔ‬หᕈᬌ⚝ࠍⴕߞߚ(⴫ 1)‫⩶ߩߡߴߔޕ‬ᩣߪᅢࠕ࡞ࠞ࡝ᕈࡃ࠴
࡞ࠬዻ⚦⩶ߣᦨ߽㜞޿⋧หᕈࠍ␜ߒߚ߇‫ߜ߁ߩߘޔ‬ߩ 1 ⩶ᩣ߇ B. pseudofirmus OF4 ᩣߩႮၮ㈩೉ߣ 100%৻⥌ߒߡ޿ߚ‫ޕ‬KU50 ᩣߣ๮ฬߒߚߎߩ⚦⩶ߪ‫ޔ‬ㆇേᕈߩ޽ࠆࠣ࡜ࡓ㓁ᕈߩ᪝⩶ߢ‫⚦ޔ‬⢩ߩ┵ߦᲣ⚦⢩߇⤘ࠄ߹ߥ޿⢩ሶᒻᚑࠍⴕ޿‫࠯࡯࡜࠲ࠞޔ‬ᵴᕈߣࠝࠠࠪ࠳࡯࠯ᵴᕈࠍ␜ߒߚ‫ߩࠄࠇߎޕ‬ᕈ⾰ߪߔߴߡ B.
pseudofirmus OF4 ᩣߩᕈ⾰ߣหߓߢ޽ߞߚߎߣ߆ࠄ‫ޔ‬
KU50 ᩣߪ B. pseudofirmus OF4 ᩣߩㄭ✼⒳ߢ޽ࠆߎ
ߣ߇ផቯߐࠇߚ‫৻ޕ‬ᣇߢ‫ޔ‬KU50 ᩣߪ Na+ߣ Li+ߦኻߔࠆᕈ⾰߇ OF4 ᩣߣߪᄢ߈ߊ⇣ߥߞߡ޿ߚ(࿑ 1)‫ޕ‬↢⢒ታ㛎ߩ⚿ᨐ߆ࠄ‫ޔ‬OF4 ᩣߪ pH 10.5 ߦ߅޿ߡ⥋ㆡ↢⢒ߦ 10 mM એ਄ߩ NaCl ࠍᔅⷐߣߒ‫ޔ‬LiCl ߦߟ޿ߡߪ 300 mM ߹ߢᷝടߒߡ߽↢⢒ߪⷰኤߐࠇߥ
߆ߞߚߩߦኻߒ‫ޔ‬KU50 ᩣߪ NaCl ߿ LiCl ࠍᷝടߒߥߊߡ߽↢⢒ߒ‫ޔ‬LiCl ߦኻߒߡߪ 100 mM ߹ߢ⠴ᕈ߇޽ߞߚ‫⚿ߩߎޕ‬ᨐߪ‫ޔ‬KU50 ᩣߩ⚦⢩ౝߩ㉄ᕈൻᯏ᭴ߦߪ OF4 ᩣ߇ⷐ᳞ߔࠆߩߣห⒟ᐲߩ Na+߿ Li+ࠍᔅⷐߣߒߥ޿ߎߣ‫ޔࠅ߹ߟޔ‬೎ߩ㓁ࠗࠝࡦࠍ೑↪
ߔࠆ߆‫ߊߚߞ߹ޔ‬೎ߩᯏ᭴߇ሽ࿷ߔࠆߎߣࠍ␜ໂߒߚ‫ޕ‬

⴫ 1. 16S rDNA Ⴎၮ㈩೉ࠍ↪޿ߚಽ㔌⩶ᩣߩหቯ
ᦨ䉅㜞䈇⋧หᕈ䉕␜䈚䈢↢‛⒳ a)න㔌䈘䉏䈢⩶ᩣᢙ

Bacillus lehensis

33 ⩶ᩣ

Bacillus polygoni

11 ⩶ᩣ

Bacillus clausii

7 ⩶ᩣ

Bacillus neizhouensis

3 ⩶ᩣ

Bacillus clarkii

1 ⩶ᩣ

Bacillus pseudofirmus

1 ⩶ᩣ

⒳߹ߢหቯߐࠇߡ޿ࠆ↢‛⒳ߩਛߢ‫ޔ‬ಽ㔌⩶ᩣߩ 16S rDNA Ⴎၮ㈩೉ߣᦨ߽㜞޿⋧หᕈࠍ␜ߒߚ 16S rDNA
Ⴎၮ㈩೉ࠍᜬߞߚ↢‛⒳‫ޕ‬a)72

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)KU50 ᩣߩ࠼࡜ࡈ࠻ࠥࡁࡓߩ⸃⺒ߣ OF4 ᩣࠥࡁࡓߣ
ߩᲧセ 2 ߟߩ B. pseudofirmus ᩣߩ↢⢒ߦ߅ߌࠆࠗࠝࡦଐ
ሽᕈߩ㆑޿ࠍ⺞ߴࠆߚ߼‫ޔ‬KU50 ᩣߩ࠼࡜ࡈ࠻ࠥࡁ
ࡓࠍ࠲ࠞ࡜ࡃࠗࠝᩣᑼળ␠࠼࡜ࠧࡦࠫࠚࡁࡒࠢࠬ࠮ࡦ ࠲ ࡯ߢ ⸃⺒ ߒ ߚ‫ ޕ‬ᓧࠄ ࠇ ߚࠥ ࡁࡓ ߩ ࠺࡯ ࠲ߪ
EDENA (http://www.genomic.ch/edena.php)4) ߢࠕ࠶࠮ࡦࡉ࡝࡯ߐࠇ‫ޔ‬1,306 ୘ߩࠦࡦ࠹ࠖࠣ߇ᓧࠄࠇߚ‫ޕ‬ᰴߦ‫ޔ‬ORF finder(http://www.ncbi.nlm.nih.gov/gorf/gorf.html)ߣ BLAST
Search (http://blast.ncbi.nlm.nih.gov/Blast.cgi)‫ߪߚ߹ޔ‬in silico MolecularCloning Genomics Edition ࠍ↪޿ߡߎࠇࠄߩࠦࡦ࠹ࠖࠣ߆ࠄផቯ਄ߩㆮવሶࠍ᛽಴ߒ‫ޔ‬⋧หᕈᬌ⚝ࠍⴕߞߚ‫ޕ‬ᓧࠄࠇߚࠕࡁ࠹࡯࡚ࠪࡦ࠺࡯
࠲ࠍ߽ߣߦ OF4 ᩣߣ KU50 ᩣߩ㆑޿ࠍ᛽಴ߒߚ‫߹ޕ‬ߕ‫ޔ‬OF4 ᩣߩ Na+ߩᓴⅣ♽ࠍ᭴ᚑߔࠆ Na ࠴ࡖࡀ࡞ߣ
Na+/H+ࠕࡦ࠴ࡐ࡯࠲࡯ࠍᲧセߒߚ‫ޕ‬Na ࠴ࡖࡀ࡞ߩㆮ
વሶ(ncbA)ߪߤߜࠄߩ⩶ᩣߦ߽ 1 ߟߛߌሽ࿷ߒ‫ߤޔ‬
ߜࠄ߽ 284 ࠕࡒࡁ㉄ᱷၮ߆ࠄߥࠆ࠲ࡦࡄࠢ⾰ࠍࠦ࡯࠼ߒߡ޿ࠆߎߣ߇ផቯߐࠇߚ‫ޕ‬284 ࠕࡒࡁ㉄ᱷၮߩ߁ߜ 6 ▎ᚲߩࠕࡒࡁ㉄ᱷၮߦ߅޿ߡ⋧㆑߇⹺߼ࠄࠇ‫ޔ‬ߘߩ߁ߜߩ 1 ▎ᚲ߇ࠗࠝࡦㆬᛯࡈࠖ࡞࠲࡯ߢ޽ࠆߎ
ߣ߇᣿ࠄ߆ߣߥߞߚ‫ޔߪߣߎߩߎޕ‬KU50 ᩣߩ Na ࠴ࡖࡀ࡞ߩࠗࠝࡦㆬᛯᕈ߇ OF4 ᩣߩ߽ߩߣ⇣ߥࠆߎߣࠍ␜ໂߒߡ޿ࠆ‫⚦ޔߚ߹ޕ‬⢩ౝߩ㉄ᕈൻߦ⋥ធ㑐ਈߔࠆਥⷐߥ Na+/H+ࠕࡦ࠴ࡐ࡯࠲࡯(MrpABCDEFGࠪࠬ࠹ࡓ)ࠍ OF4 ᩣߪ 1 ࠮࠶࠻ߒ߆ᜬߞߡ޿ߥ޿߇‫ޔ‬

KU50 ᩣߪ 2 ࠮࠶࠻଻ᜬߒߡ޿ࠆߎߣ߇᣿ࠄ߆ߣߥߞߚ‫ ߩߎޕ‬2 ߟߩ Mrp ࠪࠬ࠹ࡓߩࠗࠝࡦㆬᛯᕈߦ㆑
޿߇޽ࠆߩ߆ߤ߁߆ߪ৻ᰴ᭴ㅧ਄ߩᲧセ߆ࠄߪ᣿ࠄ
߆ߦߥߞߡ޿ߥ޿߇‫৻ޔ‬ᣇߪ OF4 ᩣߩ Mrp ࠪࠬ࠹ࡓ
ߣ 89㨪100%ߩ㜞޿⋧หᕈ߇޽ࠅ‫৻߁߽ޔ‬ᣇߪ⋧ห
ᕈ߇ 44㨪61%ߣૐ߆ߞߚߎߣ߆ࠄ‫ࡦࠝࠗޔ‬ㆬᛯᕈߦ㆑޿߇޽ߞߡ߽߅߆ߒߊߪߥ޿‫ޕ‬એ਄ߩߎߣ߆ࠄ‫ޔ‬
KU50 ᩣߪ Na+ߩขࠅㄟߺ♽ߣឃ಴♽ߩਔᣇߦ߅޿
ߡ‫ޔ‬OF4 ᩣߣߪ⋧㆑߇޽ࠅ‫ޔ‬Na+એᄖߩࠗࠝࡦࠍ(߽)ࠕ࡞ࠞ࡝ⅣႺ߳ߩㆡᔕߦ೑↪ߒߡ޿ࠆน⢻ᕈ߇⠨߃ࠄࠇߚ(࿑ 3)‫․ߩ߆߶ߩߘޕ‬ᓽߣߒߡ‫ޔ‬KU50 ᩣߩࠥࡁࡓߦߪ‫ ࡦࡒ࠲ࡆޔ‬B12 วᚑ⚻〝ߦ㑐ࠊࠆㆮવሶ⟲(cysG, cbiX, cbiL, cbiH, cbiF, cbiG, cbiD, cbiJ, cbiE,
cbiT, cbiC, cbiA, btuR)߿‫ߩ࡞࡯࠻ࠪࡁࠗޔ‬ઍ⻢ߦ㑐ࠊࠆㆮવሶ⟲(iolG, iolE, iolI, iolD, iolB, iolC, iolJ)‫ޔ‬⢩ሶߩ㉄ൻࠬ࠻࡟ࠬ⠴ᕈߦ㑐ࠊࠆㆮવሶ⟲(mnxABCDEFG)ߥߤ‫ޔ‬OF4 ᩣߦߪሽ࿷ߒߥ޿ઍ⻢⚻〝߇ሽ࿷ߔࠆߎߣ߇᣿ࠄ߆ߣߥߞߚ‫੹ޔߚ߹ޕ‬࿁ߩ࠼࡜ࡈ࠻ࠥࡁࡓ߆ࠄߪ OF4 ᩣߩᜬߟ⚦⢩ოวᚑㆮવሶ⟲(slpA, slpA2, csaA, csaB)߇⷗ߟ߆ߞߡ߅ࠄߕ‫ޔ‬OF4 ᩣߣ KU50 ᩣߩ⚦⢩⴫ጀߦ㆑޿߇޽ࠆߎߣ߇␜
ໂߐࠇߚ(࿑ 3)‫ޕ‬OF4 ᩣߩ߽ߟ SlpA ࡟ࠗࡗ࡯ߪࠕ࡞
ࠞ࡝ pH ߳ߩࠕ࠶ࡊࠪࡈ࠻ᤨߩㆡᔕߦ㊀ⷐߥᓎഀࠍ
ᜬߞߡ޿ࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ 3) ‫ޔߡߞ߇ߚߒޕ‬KU50 ᩣߩ⚦⢩⴫ጀ߇ߤߩࠃ߁ߥ᭴ㅧߢ‫߁ࠃߩߤޔ‬ߦࠕ࡞ࠞ࡝ⅣႺ߳ߩㆡᔕߦ㑐ਈߒߡ޿ࠆߩ߆ߪ⥝๧ᷓ޿ὐߢ޽ࠆ‫ޕ‬

࿑ 3. B. pseudofirmus KU50 ᩣߩࠥࡁࡓ⸃⺒߆ࠄផኤߐࠇࠆࠕ࡞ࠞ࡝ⅣႺㆡᔕᯏ᭴ߩࡕ࠺࡞
B. pseudofirmus OF4 ᩣߣ⇣ߥࠆ▎ᚲࠍ⿒޿⍫ශߣ⿒޿ᢥሼߢ␜ߒߚ‫ޕ‬OF4 ᩣߩ MrpABCDEFG ࠪࠬ࠹ࡓߣ㜞޿⋧หᕈࠍᜬߟ KU50 ᩣߩ Mrp1 ߣ Na ࠴ࡖࡀ࡞߇ Na+ߩᓴⅣߦ౒ᓎߒߡ⚦⢩ౝࠍ㉄ᕈൻߔࠆ‫ޕ‬KU50 ᩣߦߪ
೎ߩ MrpABCDEFG ࠪࠬ࠹ࡓ(Mrp2)߇޽ࠆὐߣ Na ࠴ࡖࡀ࡞ߩࠗࠝࡦㆬᛯᕈߦ㑐ࠊࠆࠕࡒࡁ㉄ᱷၮ߇ OF4 ᩣߣߪ⇣ߥߞߡ޿ߚὐ߆ࠄ‫ޔ‬೎ߩࠞ࠴ࠝࡦ(X+)ߩᓴⅣᯏ᭴ߣ౒⸶ߒߚ⚦⢩ౝ㉄ൻᯏ᭴߽ᜬߟߎߣ߇ផኤߐࠇࠆ‫ޕ‬ട߃ߡ‫⚦ޔ‬⢩⴫ጀ߇ OF4 ᩣߣߪ⇣ߥࠆߎߣ߇␜ໂߐࠇࠆ‫ޕ‬73

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)੹ᓟߩዷᦸ ᧄ⎇ⓥߢ↪޿ߚૐ Na+ၭ࿾ਛߢߩࠬࠢ࡝࡯࠾ࡦࠣ
ߦࠃߞߡᓧࠄࠇߚᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߪ‫ࡖ࠴ࡦࠝࠗޔ‬ࡀ࡞ߩࠗࠝࡦㆬᛯࡈࠖ࡞࠲࡯ㇱ૏ߦࡇࡦࡐࠗࡦ࠻ߢ
ᄌ⇣߇౉ߞߡ޿ߚ‫ߩߎޕ‬ᄌ⇣߇ታ㓙ߦ Na ࠴ࡖࡀ࡞ߩࠗࠝࡦㆬᛯᕈߦᓇ㗀ࠍਈ߃ࠆ߆ߦߟ޿ߡߪ⃻࿷⺞ᩏਛߢ޽ࠆ߇‫ࡦࠝࠗߒ߽ޔ‬ㆬᛯᕈ߇⇣ߥߞߡ޿ࠆߥ
ࠄ߫‫ޔ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ࠍ↪޿ߚᧄࠬࠢ࡝࡯࠾ࡦࠣ
♽߇೎ߩࠗࠝࡦߩャㅍ૕ߩਛߦ޽ࠆࠗࠝࡦㆬᛯㇱ૏ߩផቯߦ߽ᵴ↪ߢ߈ࠆ߆߽ߒࠇߥ޿‫੹ޔߚ߹ޕ‬࿁න㔌ߐࠇߚ KU50 ᩣߩࠥࡁࡓਛߩ᭽‫․ߥޘ‬ᓽ߇‫ޔ‬ታ㓙ߦ⚦⢩ౝߩ㉄ᕈൻ߿ Li+⠴ᕈߦ㑐ਈߒߡ޿ࠆߩ߆ߤ
߁߆ߦߟ޿ߡߪ‫੹ޔ‬ᓟ‫ߩࠄࠇߎޔ‬ㆮવሶߩᰳ៊ᩣ߿
ᄌ⇣ᩣࠍ᭴▽ߔࠆߎߣߢ⏕⹺ߔࠆᔅⷐ߇޽ࠆ‫ߩߎޕ‬ࠃ߁ߥ⎇ⓥߩㆀⴕࠃߞߡ‫ޔ‬ᅢࠕ࡞ࠞ࡝ᕈ⚦⩶ߩᣂߚߥࠕ࡞ࠞ࡝ⅣႺㆡᔕᯏ᭴߇᣿ࠄ߆ߦߥࠆߎߣ߇ᦼᓙ
ߐࠇࠆ‫ޕ‬

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⻢ㄉ ᧄ⎇ⓥߪ᧲ᵗᄢቇ ᐔᚑ 23 ᐕᐲ ․೎⎇ⓥߩᡰេ
ࠍฃߌߡታᣉߐࠇ߹ߒߚ‫ޕ‬ෳ⠨ᢥ₂1) Fujinami S. and Fujisawa M. 2010. Industrial
applications of alkaliphiles and their enzymes –past, present and future. Environ. Technol. 31:
845~856.
2) Gilmour R. and Krulwich T. A. 1997. Construction
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Bacillus firmus OF4 with a disrupted cta operon
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Two-dimensional gel electrophoresis analyses ofpH-dependent protein expression in facultatively

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Journal of Japanese Society for Extremophiles (2012) Vol.11, 75-82

㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Arakawa K.

Comparative metabolomics of anhydrobiosis in tardigrade
Ramazzottius varieornatusᲧセࡔ࠲ࡏࡠࡒࠢࠬߦࠃࠆ࡛ࠦ࠸࠽ࠢࡑࡓࠪੇ⌁ᯏ᭴ߩ⸃᣿
ᘮᙥ⟵Ⴖᄢቇవ┵↢๮⑼ቇ⎇ⓥᚲ
⨹Ꮉ ๺᥍Institute for Advanced Biosciences, Keio University, 5322 Endo, Fujisawa, Kanagawa, 252-0882, JAPAN
Corresponding author: Kazuharu Arakawa, gaou@sfc.keio.ac.jp
Tel/Fax: 0466-47-5099
Received: Nobember 6, 2011 / Accepted: December 16, 2011
Abstract Limno-terrestrial tardigrades can withstandalmost complete desiccation through a mechanism called
anhydrobiosis, and several of these species have beenshown to survive the most extreme environments throughexposure to space vacuum. Molecular mechanism forthis tolerance has so far been studied in many
anhydrobiotic metazoans, leading to the identification ofseveral key molecules such as the accumulation andvitrification of trehalose as well as the expression of LEA
proteins to prevent protein aggregation. On the other
hand, the understanding of comprehensive molecularmechanisms and regulation machinery of metabolic
compounds during anhydrobiosis is yet to be explored. In
this review, I report and summarize the recent progress inthe comprehensive metabolome analysis using thetardigrade Ramazzottius varieornatus, which is apotential model species for anhydrobiosis. In order toanalyze the metabolic changes in the active anddehydrated states, metabolome is measured in bothconditions using two types of high-throughput mass
spectrometry (MS) systems, liquid chromatography
time-of-flight MS (LC-TOFMS) for lipids and sugars
and capillary electrophoresis TOFMS (CE-TOFMS) for
primary metabolite. Increase, but no significant
accumulation of trehalose in this species was confirmed,
suggesting a more complex mechanism foranhydrobiosis in comparison to other metazoans. Whilechanges in gene expression profiles are limited in
between active and tun states, dynamic changes wereobserved in the metabolism of this species in response to
desiccation. Changes in the metabolic profiles suggest
complex intracellular responses to oxidative and osmotic
stress.

Keywords: Anhydrobiosis, metabolomics, tardigrade,
cryptobiosis, Ramazzottius varieornatus, oxidative stressࠠ࡯ࡢ࡯࠼: ੇ⌁‫ࠢޔࠪࡓࡑࠢޔࠬࠢࡒࡠࡏ࠲ࡔޔ‬࡝ࡊ࠻ࡃࠗࠝࠪࠬ‫㉄ޔࠪࡓࡑࠢ࠽࠸࡛ࠦޔ‬ൻࠬ࠻࡟ࠬ
1. ╙ਃߩ↢๮⁁ᘒ‫ޠࠬࠪࠝࠗࡃ࠻ࡊ࡝ࠢޟ‬ ↢๮ߣߪ૗߆‫ߩߎޕ‬ⓥᭂߩ໧޿߳ߩ࿁╵ߪ↢‛ቇ⊛࡮‛ℂቇ⊛┙႐ߥߤ߆ࠄ➅ࠅ㄰ߒ⹜ߺ߇޽ࠆ߽ߩ
ߩଐὼߣߒߡືቇߩၞࠍ಴ߥ޿߇‫ޔ‬ᢔㅺ᭴ㅧ⛽ᜬߩߚ߼ߦઍ⻢߇ਇนᰳߢ޽ࠆߣ޿߁ὐߪ౒ㅢ⹺⼂ߩ৻ߟߢ޽ࠆߣᕁࠊࠇࠆ‫᦭߽ߦࠅ߹޽ޕ‬ฬߥࠪࡘ࡟࠺ࠖ
ࡦࠟ࡯ߦࠃࠆ⼏⺰ߢߪ‫ޠ࡯ࡇࡠ࠻ࡦࠛߩ⽶ޟߪ‛↢ޔ‬ࠍ⛘߃ߕขࠅㄟ߻ߎߣߦࠃߞߡ↢๮ᵴേࠍ⛽ᜬߒ‫ޔ‬‫ᦨޟ‬ᄢࠛࡦ࠻ࡠࡇ࡯‫⁁ޠ‬ᘒߢ޽ࠆ‫ޟ‬ᱫ‫ࠄ߆ޠ‬ㅏࠇߡ޿ࠆ‫ߣޔ‬ㅀߴࠄࠇߡ޿ࠆ 26‫ߥ߁ࠃߩߎޕ‬ઍ⻢ࠍゲߣߒߚਇนㅒߥ↢ߣᱫߩ᭎ᔨߪᐢߊ৻⥸ߦฃߌ౉ࠇࠄ
ࠇߡ޿ࠆ߽ߩߢ޽ࠆ߇‫ߪ‛↢ޔ‬Ᏹߦ଀ᄖߩቲᐶߢ޽
ࠆ‫ޕ‬ਥߦ㒽↢ߩ㕖⣄ᬁേ‛ࠍਛᔃߣߒߚ޿ߊߟ߆ߩ
↢‛⒳ߪ‫ߥ߹ߑ߹ߐޔ‬ⅣႺᄌേ㧔ੇ῎࡮ૐ᷷࡮ૐ㉄⚛࡮ᶐㅘ࿶࡮Ქ⚛ߥߤ㧕ߦࠃߞߡࠢ࡝ࡊ࠻ࡃࠗࠝࠪ
ࠬߣ๭߫ࠇࠆήઍ⻢ߩભ⌁⁁ᘒߦ⒖ⴕߔࠆߎߣ߇ߢ
߈‫ޔ‬ⅣႺߩᡷༀߦࠃߞߡㅦ߿߆ߦ↢๮ᵴേࠍౣ㐿ߔࠆߎߣ߇ߢ߈ࠆ 17‫ߪࠬࠪࠝࠗࡃ࠻ࡊ࡝ࠢޔߡߞࠃޕ‬นㅒ⊛ߥઍ⻢ᵴേߩ஗ᱛߢ޽ࠅ‫ޔ‬ᕡᏱ⊛ߥઍ⻢ᵴേ߇⷗ࠄࠇࠆ‫ޠ↢ޟ‬‫ޔ‬ਇนㅒ⊛ߥઍ⻢ᵴേߩ஗ᱛߢ޽ࠆ
‫ޟ‬ᱫ‫ޠ‬ߦട߃ߚ╙ਃߩ↢๮⁁ᘒߣ૏⟎ߠߌࠄࠇࠆ 2‫ޕ‬ߟ߹ࠅ‫ߩࠬࠪࠝࠗࡃ࠻ࡊ࡝ࠢޔ‬ಽሶᯏ᭴ߦ㑐ߔࠆ⎇ⓥߦࠃߞߡ‫↢ޔ‬๮ᵴേߣߪ૗߆‫ޔ‬ਇนㅒߥ↢๮ᵴേߩ஗ᱛ⁁ᘒߢ޽ࠆ‫ޟ‬ᱫ‫ߪߣޠ‬૗߆‫ࠍޔ‬᣿ࠄ߆ߦߒߡ
޿ߊߎߣ߇ߢ߈ࠆߣ⠨߃ࠄࠇࠆ‫ޕ‬75

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2) ࠢ࡝ࡊ࠻ࡃࠗࠝࠪࠬߩಽሶᯏ᭴ߪᧂ⍮ߩㇱಽ߇ᄙ޿߇‫ߩߘޔ‬ਛߢ⃻࿷ᦨ߽⎇ⓥ߇ㅴࠎߢ޿ࠆߩ߇ੇ⌁㧔ࠕࡦࡂࠗ࠼ࡠࡃࠗࠝࠪࠬ㧕ߢ޽ࠆ 4, 9, 31, 32‫ࡊ࡝ࠢޕ‬࠻ࡃࠗࠝࠪࠬ߇⺃ዉߐࠇࠆⅣႺᄌേߪၮᧄ⊛ߦṁᇦߢ޽ࠆ᳓ߦᓇ㗀ࠍਈ߃ࠆ߽ߩ߫߆ࠅߢ޽ࠆ߇‫⌁ੇޔ‬ߢߪࠃࠅ⋥ធ⊛ߦ‫ޔ‬๟࿐ߩੇ῎ߦࠃߞߡ↢૕ౝߩ᳓
ಽࠍᢙ%ߦ߹ߢᷫዋߐߖ 5‫ߩߎޔ‬㊂ߩ᳓ಽ㊂ߢߪ⚦⢩ౝߩಽሶ߇᳓๺ߐࠇߥ޿ߚ߼‫ޔ‬ήઍ⻢⁁ᘒ߳ߣ⒖ⴕߔࠆ‫⌁ੇޕ‬ਛߪ↢๮ᵴേ߇஗ᱛߒߡ޿ࠆߚ߼ߦᢙᐕ
߆ࠄᢙචᐕߩ㐳ᦼߦᷰߞߡ଻ሽ߇น⢻ߢ޽ࠅ‫⌀ޔ‬ⓨਛߥߤߦ⟎ߊ੐ߦࠃߞߡ㉄ൻࠍᅹߍࠆߎߣ߇ߢ߈ࠇ
߫ඨ᳗ਭ⊛ߥ଻ሽ߽น⢻ߢ޽ࠆߣ⠨߃ࠄࠇߡ޿ࠆ 14,22‫⁁⌁ੇޔߦࠄߐޕ‬ᘒߢߪ⚦⢩ਛߦ߶ߣࠎߤ᳓߇ሽ
࿷ߒߥ޿ߚ߼ߦߐ߹ߑ߹ߥᭂ㒢ⅣႺ߳ߩ⠴ᕈ߇޽ࠆߎߣ߇⍮ࠄࠇߡ߅ࠅ‫ߪߢࠪࡓࡑࠢ߫߃଀ޔ‬-200͠߆
ࠄ 100͠⒟ᐲߩ᷷ᐲ‫⌀ޔ‬ⓨ߆ࠄ 7.5GPa ߩ࿶ജ‫࡞ࠕޔ‬ࠦ࡯࡞࡮⥇ൻࠛ࠴࡞࡮ࠕ࠮࠻࠾࠻࡝࡞ߥߤߩ⮎ຠ‫ޔ‬6,000Gy ߩǫ✢ߥߤߩ⁁ᘒߦ⟎߆ࠇߚᓟߦ߽ๆ᳓ߦ
ࠃߞߡㅦ߿߆ߦᵴേࠍౣ㐿ߢ߈ࠆ 32, 33‫ߥ߁ࠃߩߎޕ‬ᭂ㒢ⅣႺ⠴ᕈߩᦨ߽⽎ᓽ⊛ߥ߽ߩ߇ Jönsson ࠄߦࠃߞߡႎ๔ߐࠇߚቝቮⓨ㑆߳ߩ᥸㔺ታ㛎ߢ޽ࠅ‫ޔ‬ഀวߪዋߥ޿߽ߩߩ‫ޔ‬㧝㧜ᣣ㑆ߩ⌀ⓨ࡮⿥ૐ᷷࡮ቝቮ✢߳ߩ᥸㔺ᓟߦࠢࡑࡓࠪߩੇ⌁߆ࠄߩᓳᏫ߇⷗ࠄࠇߡ
޿ࠆ 15‫ߪߢࠬࠪࠝࠗࡃ࠻ࡊ࡝ࠢޕ‬ᅤ૗ߦߒߡήઍ⻢⁁ᘒࠍ᜽ߺߥ߇ࠄ߽↢૕ߩ᭴ᚑⷐ⚛ࠍߎߩࠃ߁ߦ㗎ᒝߦ⛽ᜬߒ⛯ߌࠄࠇߡ޿ࠆߩ߆‫ޔߡߒߘޔ‬ᅤ૗ߦߒߡ↢๮ᵴേߩౣ㐿ࠍน⢻ߦߒߡ޿ࠆߩ߆‫ߩߘޕ‬ಽሶᯏ᭴ࠍ⸃߈᣿߆ߔߎߣߪ‫↢ޔ‬๮ᵴേߩᧄ⾰ߦㄼࠆㄭ㆏ߣᚑࠅ߁ࠆ‫ޕ‬ ⚦⢩ߩ߶߷ቢోߥ⣕᳓ߣ޿߁഍⊛ߥⅣႺᄌേ߆ࠄ
ߩ↢๮ᵴേߩౣ㐿ࠍน⢻ߦߔࠆߚ߼ߦߪ‫ޔ‬૗ࠃࠅ߽ੇ⌁ᤨߦ↢૕⤑߿ DNA‫⚦ߩߤߥ⾰ࠢࡄࡦ࠲ߡߒߘޔ‬⢩ౝߩฦⷐ⚛ࠍ‫ޔ‬ౣ᳓๺ᤨߦߘࠇࠄߩᯏ⢻ࠍ⛽ᜬߢ߈ࠆࠃ߁ੇ῎ߦࠃࠆ࠳ࡔ࡯ࠫ߆ࠄ଻⼔ߔࠆߎߣ߇⢄ⷐߢ޽ࠆ‫⼔଻ߥ߁ࠃߩߎޕ‬ᯏ᭴ߣߒߡ‫ࠕࡒ࠹࡞ࠕޔ‬߿ࡀࡓ࡝࡙ࠬ࡝ࠞߥߤߦ߅޿ߡߪ㕖ㆶర♧ߢ޽ࠆ࠻࡟ࡂࡠ࡯ࠬߩ⫾Ⓧ߇㗼⪺ߢ޽ࠅ‫῎ੇߦᤨ⌁ੇޔ‬㊀㊂
ߩታߦ 10ޯ20%ߦ߹ߢ⥋ࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ 30‫ޕ‬࠻࡟ࡂࡠ࡯ࠬߪ↢૕ౝߢ᳓ࠍ⟎឵ߒ‫⤑߿ࠢࡄࡦ࠲ޔ‬ࠍ଻⼔ߒ‫ޔߚ߹ޔ‬᛫㉄ൻ૞↪ࠍᜬߟ੐ߥߤ߇⍮ࠄࠇ
ߡ޿ࠆ 23‫ࡂ࡟࠻ߪߡ޿߅ߦࠞ࡝࡙ࠬ࡝ࡓࡀޔߦ․ޕ‬ࡠ࡯ࠬߩࠟ࡜ࠬൻߦࠃࠆ଻⼔߇ᔅ㗇ߢ޽ࠅ‫ޔ‬ㅦ޿ੇ῎ߢߪ࠻࡟ࡂࡠ࡯ࠬߩ⫾Ⓧ㊂߇ਇචಽߦߥߞߡߒ߹
޿⯃↢߇ߢ߈ߥ޿ߎߣ߇⹦⚦ߥ⸃ᨆߦࠃߞߡ␜ߐࠇ
ߡ޿ࠆ 24‫ߩߘޕ‬ᓟ‫߇ࠬ࡯ࡠࡂ࡟࠻߽ߡ޿߅ߦ⯻✢ޔ‬ੇ⌁ᤨߩ⤑᭴ㅧߩ⛽ᜬߦᔅ㗇ߢ޽ࠆߎߣ߇␜ߐࠇࠆ
ߥߤ 6‫ߦ․ޔ‬ㆃ޿ੇ⌁ߦ߅޿ߡߪ࠻࡟ࡂࡠ࡯ࠬ߇ߘ
ߩಽሶᯏ᭴ߩ㎛ߣߥߞߡ޿ࠆ‫ޕ‬

2.ᄙ᭽ߥੇ⌁ᯏ᭴ ੇ⌁ߪࡃࠢ࠹࡝ࠕ߿㉂Უߥߤߩන⚦⢩↢‛‫ߒߘޔ‬ߡࡢࡓࠪ‫࡙࡝ࡓࡀޔࠕࡒ࠹࡞ࠕޔࠪࡓࡑࠢޔ⯻✢ޔ‬ࠬ࡝ࠞߥߤߩᓸዊേ‛ߦ᏷ᐢߊ⷗ࠄࠇ 31‫৻ޔ‬ቯએਅߩᄢ߈ߐߩ↢‛ߦ߅޿ߡߪ๟ㄝⅣႺߩੇ῎ߦ㓙ߒߡߪ‛ℂ⊛ߥ⒖േ߿⣕᳓ߩ࿁ㆱߢߪߥߊੇ῎⠴ᕈࠍᜬߟߣ޿߁ᚢ⇛߇ㅴൻ⊛ߦ᦭ലߢ޽ߞߚߎߣ߇ુ߃ࠆ‫ޕ‬৻ᣇߢ‫⌁ੇޔ‬⒖ⴕߩಽሶᯏ᭴⥄૕ߪ޽ࠆ⒟ᐲᄙ᭽ߦ‫ޔ‬ߐࠄߦ޿ߊߟ߆ߩᯏ᭴ߦ㑐ߒߡߪㅴൻ⊛ߦ⁛┙ߦ₪ᓧߐࠇߡ޿ࠆߣ⠨߃ࠄࠇࠆ‫࡞ࠕ߿ࠪࡓࡢޔ߫߃଀ޕ‬࠹ࡒࠕߢߪ⠴ᕈෆ‫ߪߢࠞ࡝࡙ࠬ࡝ࡓࡀޔ‬ᐜ⯻‫⯻✢ޔ‬ߢߪ⠴ᕈᐜ⯻ߥߤ․ቯߩ⊒↢ࠬ࠹࡯ࠫߦ߅޿ߡߩߺ
ੇ⌁⒖ⴕ߇น⢻ߢ޽ࠆߩߦኻߒ‫ߪߢࠪࡓࡑࠢޔ‬ෆ࡮ᐜ૕࡮ᚑ૕ߩోࠬ࠹࡯ࠫߢߩੇ⌁߇น⢻ߢ޽ࠆ 10‫ޕ‬߹ߚ‫ߦ⌁ੇߪߢࠪࡓࡑࠢޔ‬㓙ߒߡり૕ࠍᮻߩࠃ߁ߥ
ᒻߦ❗߼⴫㕙Ⓧࠍ❗ዊߐߖࠆ tun ᒻᚑ߇ⴕࠊࠇࠆ 20‫ޕ‬ߐࠄߦ‫ߦ⌁ੇޔ‬ᔅⷐߥᤨ㑆ߦ㑐ߒߡ߽‫ޔ‬㜞޿Ḩᐲߢᢙᣣ㑆߆ߌߡੇ῎ߔࠆߎߣ߇ਇนᰳߥࡀࡓ࡝࡙ࠬ࡝
ࠞߩࠃ߁ߥㆃ޿ੇ⌁⒖ⴕࠍߔࠆࠣ࡞࡯ࡊߣ 3‫ߋߔޔ‬ߦ⋧ኻḨᐲ 0%ߦㄭ޿ࠃ߁ߥᭂ߼ߡੇ῎ߒߚ⁁ᘒߦ⟎߆ࠇߡ߽ㅦ߿߆ߥੇ⌁⒖ⴕ߇น⢻ߥ৻ㇱߩ✢ᒻേ‛߿ࠢࡑࡓࠪߩࠃ߁ߥࠣ࡞࡯ࡊߦੑಽߐࠇࠆߎߣ߇
᣿ࠄ߆ߦߥߞߡ߈ߡ޿ࠆ 27‫ޕ‬76 ৻ᣇ‫ߦࠬ࡯ࡠࡂ࡟࠻ޔ‬ଐሽߒߥ޿ੇ⌁ᯏ᭴ߩሽ࿷
߽ᰴ‫ߣޘ‬᣿ࠄ߆ߦߥߞߡ߈ߡ޿ࠆ‫ࠬ࡯ࡠࡂ࡟࠻ޕ‬วᚑ㉂⚛ߣ࠻࡟ࡂࡠ࡯ࠬࠍ⚦⢩ౝߦ߽ߚߥ޿߇㜞޿ੇ⌁⢻ജࠍ␜ߔࡢࡓࠪ߇ႎ๔ߐࠇ 18‫ࠬ࡯ࡠࡂ࡟࠻ޔ‬วᚑ㉂⚛ߩࡁ࠶ࠢࠕ࠙࠻ታ㛎ߦࠃߞߡ‫ޔ‬㉂Უߢߪੇ῎⠴ᕈߦ࠻࡟ࡂࡠ࡯ࠬ߇ᔅ㗇ߢߪߥ޿ߎߣ߇␜ߐࠇߚ‫ޕ‬߹ߚ‫ޔ‬Ყセ⊛ㅦ޿ੇ⌁߇น⢻ߥࠢࡑࡓࠪߦ߅޿ߡߪ‫ޔ‬
ᄙߊߩ⒳ߦ߅޿ߡੇ⌁ᤨߦ࠻࡟ࡂࡠ࡯ࠬ߇ᢙ୚ߦჇ
ടߔࠆ߽ߩߩ‫ߩߘޔ‬㊂ߪੇ῎㊀㊂ߩ 1%ߦ߽ḩߚߕ
ߎߩ㊂ߢߪ᳓ಽሶߩ⟎឵߿ࠟ࡜ࠬൻߦࠃࠆ⚦⢩ߩ଻⼔ߪ࿎㔍ߢ޽ࠆߎߣ‫ࠪࡓࡑࠢ࠾ࠝޔߦࠄߐޔ‬(Milnesium tardigrada)ߦ߅޿ߡߪߘ߽ߘ߽⚦⢩ౝ࠻࡟ࡂࡠ࡯ࠬ߇ᬌ಴ߢ߈ߥ޿ߎߣߥߤ߇᣿ࠄ߆ߣߥߞ
ߚ 12‫ޕ‬ ߛ߇‫ޔ‬ㆃ޿ੇ⌁ߢ޽ࠇ߫࠻࡟ࡂࡠ࡯ࠬଐሽߢ‫ޔ‬ㅦ޿ੇ⌁ߢ޽ࠇ߫㕖ଐሽ߆ߣ޿߃߫⹤ߪߘ߁න⚐ߢߪߥ޿‫ ⯻✢ޕ‬Panagrolaimus superbus ߪ޿߈ߥࠅ⋧ኻḨ
ᐲ 0%ߦ⟎߆ࠇߡ߽ੇ⌁߇น⢻ߢ޽ࠆ߇‫ߩߎޔ‬⒳ߢ
ߪⅣႺᄌേߦ㑐ࠊࠄߕᏱߦੇ῎㊀㊂ߩ 10%߶ߤߩ࠻
࡟ࡂࡠ࡯ࠬࠍ⚦⢩ౝߦḰ஻ߔࠆߎߣߢㅦ޿ੇ⌁ߦㆡᔕߒߡ޿ࠆ 27‫ࠬ࡯ࡠࡂ࡟࠻ޔࠅ߹ߟޕ‬㕖ଐሽߢㅦ޿
ੇ⌁߇น⢻ߥࠢࡑࡓࠪߦ߅޿ߡߪ․ቯߩಽሶߦᒝߊ
ଐሽߖߕߦ߅ߘࠄߊⶄᢙߩⷐ⚛߇㑐ਈߒ‫ޔ‬ᕡᏱ⊛ߦੇ⌁ߦ஻߃ߡ޿ࠆน⢻ᕈ߇㜞޿‫ޕ‬
3. ࠢࡑࡓࠪߩੇ⌁ ࠢࡑࡓࠪੇ⌁ᯏ᭴ߩ⸃ᨆߪ‫⃻ޔ‬࿷㘺⢒♽߇⏕┙ߐࠇ ߡ ޿ ࠆ ࠝ ࠾ ࠢ ࡑ ࡓ ࠪ 29 ߣ ࡛ ࠦ ࠸ ࠽ ࠢ ࡑ ࡓ ࠪ(Ramazzottius varieornatus) 13 ߩ㧞⒳ࠍਛᔃߦㅴ߼ࠄ
ࠇߡ߅ࠅ‫ࠆߌ߅ߦ⌁ੇߪߕ߹ޔ‬ಽሶᄌേࠍࠥࡁࡒࠢ
ࠬ‫ࠍࠬࠢࡒࠝ࠹ࡠࡊޔࠬࠢࡒ࠻ࡊ࡝ࠢࠬࡦ࡜࠻ޔ‬ᆎ߼ߣߒߚᚲ⻐‫✂ߡߞࠃߦ࠴࡯ࡠࡊࠕߩޠࠬࠢࡒࠝޟ‬⟜⊛ߦᝒ߃ࠆߎߣ߇⹜ߺࠄࠇߡ޿ࠆ 20‫߹ߪߢ⁁⃻ޕ‬ߛઁߩ↢‛ߦ߽౒ㅢߦ⷗ࠄࠇࠆੇ῎ࠬ࠻࡟ࠬߦ㑐ㅪ

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ᄖߦ߽ HSP90, HSP60, HSP40, HSP20, GroES, GrpE ࡈࠔࡒ࡝࡯ߥߤ߇ሽ࿷ߒߡ޿ࠆߎߣ߇⏕⹺ߐࠇߡ޿ࠆ‫ޕ‬ߒߚࡄࠬ࠙ࠚࠗߩត⚝ߦ⇐߹ߞߡ޿ࠆ߽ߩߩ‫ࠇߎޔ‬߹ߢߩ࠼ࠗ࠷ߩࠣ࡞࡯ࡊߦࠃࠆࠝ࠾ࠢࡑࡓࠪߩ৻ㅪ
ߩ࠻࡜ࡦࠬࠢ࡝ࡊ࠻࡯ࡓ෸߮ࡊࡠ࠹ࠝ࡯ࡓ⸃ᨆߢߪ
ੇ⌁ߦ߅ߌࠆᐢ▸࿐ߥ↢૕ౝࡄࠬ࠙ࠚࠗ෸߮࠲ࡦࡄ
ࠢߩ㑐ਈ߇᣿ࠄ߆ߦߥࠅߟߟ޽ࠆ 7, 19, 25‫⚦ޔ߫߃଀ޕ‬⢩⤑਄ߦሽ࿷ߒ᳓ߩឃ಴ߦ㑐ࠊߞߡ޿ࠆߣ⠨߃ࠄࠇࠆࠕࠢࠕࡐ࡝ࡦ㧔․ߦ AQ4㧕‫ޔ‬MutS ߿ RAD51 ߥߤ
ߩ DNA ୃᓳ㑐ㅪߩࡄࠬ࠙ࠚࠗ‫㉄ޔ‬ൻࠬ࠻࡟ࠬᔕ╵ߦ㑐ࠊࠆ Super Oxide Dismutase (Mn)߿ Glutathionetransferase ߩࡄࠬ࠙ࠚࠗߥߤ߇ੇ⌁ᤨߩ⊒⃻੣ㅴ߆ࠄߘߩ㑐ਈ߇␜ໂߐࠇߡ޿ࠆ‫߿ࠬ࡟࠻ࠬ῎ੇޕ‬ᶐㅘ࿶ࠬ࠻࡟ࠬᤨߦߪ DNA ߩ៊்߿↢૕࠲ࡦࡄࠢߩ㉄ൻࠍᒁ߈⿠ߎߔࡈ࡝࡯࡜ࠫࠞ࡞߇↥಴ߐࠇࠆߎߣ߇
⍮ࠄࠇߡ߅ࠅ‫㉄ߡ޿߅ߦ⌁ੇޔ‬ൻࠬ࠻࡟ࠬᔕ╵ߪ⚦⢩ߩᯏ⢻⛽ᜬߣ޿߁ⷰὐ߆ࠄਛᔃ⊛ᓎഀࠍᜂߞߡ޿
ࠆߣ⠨߃ࠄࠇࠆ‫ޔߦࠄߐޕ‬⣕᳓ߦࠃࠆ࠲ࡦࡄࠢߩಝ
❗ࠍ᛼ߐ߃ࠆߚ߼‫ߪߢ‛↢ߩઁޔ‬ਥߦᾲ߿ૐ㉄⚛ࠬ࠻࡟ࠬߦ߅޿ߡ⊒⃻߇⷗ࠄࠇࠆࠬ࠻࡟ࠬᔕ╵ဳಽሶࠪࡖࡍࡠࡦߥߤߩ⊒⃻߇ࠢࡑࡓࠪߩੇ⌁ᤨߦߪ㗼⪺ߢ޽ࠆ‫⚦ߦ․ߪߡ޿߅ߦࠪࡓࡑࠢ࠾ࠝޕ‬⢩⾰ਛߢߩ࠲ࡦࡄࠢߩᱜߒ޿ࡈࠜ࡯࡞࠺ࠖࡦࠣࠍഥߌࠆᾲ࡚ࠪ
࠶ࠢ࠲ࡦࡄࠢ(HSP)70 ߩ⊒⃻ᄌേ߇⷗ࠄࠇ‫ࠇߘޔ‬એ ੇ⌁ߦ߅޿ߡ㉄ൻࠬ࠻࡟ࠬߣห╬ߦ㊀ⷐߥ↢૕଻⼔૞↪ߣߒߡ࠲ࡦࡄࠢߩಝ❗ᛥ೙߇޽ࠅ‫߼ߚߩߘޔ‬ߦ ߶ ߣ ࠎ ߤ ߩ ੇ ⌁ ࠍ ⴕ ߁ ↢ ‛ ߇ LEA (Late
Embryogenesis Abundant)ߣ๭߫ࠇࠆ࠲ࡦࡄࠢࠍᜬߟ
11‫ߣࠬ࡯ࡠࡂ࡟࠻ߩߎޕ‬ਗࠎߢੇ⌁ᯏ᭴ߩ⺑᣿ߦᰳ߆ߖߥ޿࠲ࡦࡄࠢߪ߽ߣ߽ߣᬀ‛ߩ⒳ሶߢ⊒⷗ߐࠇ‫ޔ‬ߘߩ⹦⚦ߥಽሶᯏ᭴ߦߪᧂ⍮ߩὐ߽ᄙ޿߽ߩߩ‫ੇޔ‬῎ࠬ࠻࡟ࠬߦ߅޿ߡߪ⚦⢩଻⼔ߩߐ߹ߑ߹ߥ㕙ߢᓎഀࠍᨐߚߔਁ⢻ㆬᚻߢ޽ࠆ‫ޔߕ߹ޕ‬LEA ࠲ࡦࡄࠢߪ
ᭂ߼ߡⷫ᳓ᕈ߇㜞ߊ‫ޔ‬㐳ᤨ㑆ߩᾚᴣࠍ⚻ߡ߽นṁᕈ࡮᛫ಝ❗ᕈࠍ⛽ᜬߢ߈ࠆߚ߼‫ߦᤨࠬ࡟࠻ࠬ῎ੇޔ‬࠲ࡦࡄࠢߩಝ❗ᛥ೙ߦ㑐ࠊࠆߎߣ߇␜ໂߐࠇߡ޿ࠆ‫ޕ‬߹ߚ‫ߪࠢࡄࡦ࠲ߩߎޔ‬᳓๺ᤨߦߪ᭴ㅧࠍߣࠄߥ޿߇‫ޔ‬ੇ῎ᤨߦߪ․ᓽ⊛ߥǩࡋ࡝࠶ࠢࠬ᭴ㅧࠍᒻᚑߒ‫ޔ‬ಽሶࠪ࡯࡞࠼ߣߒߡߩᯏ⢻ࠍ␜ߒ‫ࡦࠦ࠻ࡒߪߦࠄߐޔ‬࠼࡝ࠕߩ⤑ߩ଻⼔߿࠻࡟ࡂࡠ࡯ࠬߥߤߩ♧ߩࠟ࡜ࠬൻࠍ቟ቯൻߔࠆ૞↪߹ߢ߇ႎ๔ߐࠇߡ޿ࠆ‫ࠢ࠾ࠝޕ‬ࡑࡓࠪߩࡊࡠ࠹ࠝࡒࠢࠬߢߪੇ⌁ߦ߅ߌࠆ LEA ࠝ࡯࠰ࡠࠣߩ⺃ዉ߇ⷰኤߐࠇߡ߅ࠅ‫⌁ੇߩࠪࡓࡑࠢޔ‬ߦ߅޿ߡ߽ LEA ߇㑐ਈߒߡ޿ࠆน⢻ᕈ߇޽ࠆ‫ޕ‬

࿑ 1 㧦ᵴേ⁁ᘒߩ࡛ࠦ࠸࠽ࠢࡑࡓࠪ
ᄌൻࠍ⿠ߎߒߡ޿ࠆߩ߆ࠍ⺞ߴࠆ਄ߢߩࡕ࠺࡞↢‛ߣߒߡㆡߒߡ޿ࠆߣ⠨߃ࠄࠇࠆ‫ࠪࡓࡑࠢ࠽࠸࡛ࠦޕ‬ߪᣢߦࠢࡑࡓࠪߣߒߡߪೋߣߥࠆ⚂ 60Mbp ߩ㜞♖ᐲߥ࠼࡜ࡈ࠻ࠥࡁࡓ߇⸃⺒ߐࠇᯏ⢻ࠕࡁ࠹࡯࡚ࠪࡦ
߇ㅴࠎߢ޿ࠆ 16‫ޔߡ޿߅ߦࠪࡓࡑࠢ࠽࠸࡛ࠦߩߎޕ‬ᦨㄭጊญࠄߦࠃߞߡੑߟߩᣂⷙߥ㜞นṁᕈ࠲ࡦࡄࠢࡈ ࠔ ࡒ ࡝ ࡯ CAHS/SAHS (Cytoplasmic/Secretory
Abundant Heat Soluble)߇⊒⷗ߐࠇߚ 34‫ߩߎޕ‬ੑߟߩᣂⷙ࠲ࡦࡄࠢࡈࠔࡒ࡝࡯ߪ LEA ߩࠃ߁ߥ᛫ಝ❗ᕈߣ‫ߩᤨ῎ੇޔ‬ǩࡋ࡝࠶ࠢࠬ᭴ㅧᄌൻࠍ␜ߒ‫✭ޔ‬ᱠേ‛㐷ߦ߅޿ߡ଻ሽߐࠇ‫ࡓࡁࠥޔ‬ਛߦⶄᢙࠦࡇ࡯ሽ࿷ ⃻࿷ᚒ߇࿖ߩࠢࡑࡓࠪ⎇ⓥࠣ࡞࡯ࡊߢߪ‫ޔ‬ၳᎹࠄߦࠃߞߡർᶏ㆏߆ࠄណขߐࠇ‫ޔ‬቟ቯߒߚ㘺⢒♽ߩ߽ߣන♽⛔ൻ(YOKOZUNA-1)ߐࠇߚ࡛ࠦ࠸࠽ࠢࡑࡓ
ࠪࠍ⸃ᨆኻ⽎ߣߒߡ޿ࠆ 13‫ߘߪࠪࡓࡑࠢ࠽࠸࡛ࠦޕ‬ߩฬߩㅢࠅ‫ޔ‬㘺⢒ߩኈᤃߐ߿ᭂ㒢ⅣႺ⠴ᕈ‫ߡߒߘޔ‬ੇ⌁⒖ⴕᤨ㑆ߩ⍴ߐߥߤߦ߅޿ߡઁߩࠢࡑࡓࠪࠍಒ
㚧ߔࠆᕈ⾰ࠍ౗ߨឥ߃ߡ߅ࠅ‫ߩߘޔ‬ᭂ߼ߡᕆㅦߥੇ
⌁⒖ⴕߦ߅޿ߡߪ‫ߥ߁ࠃࠆࠇࠄ⷗ߦ‛↢⌁ੇߩઁޔ‬ㆮવሶ⊒⃻ߩᄌേ߇߶ߣࠎߤ⷗ࠄࠇߥ޿‫ޔߡߞࠃޕ‬ߎߩ⒳ߪ‫⚦ߡߒߦ߁ࠃߩߤޔ‬⢩ౝߩ᭴ᚑⷐ⚛ߩᄢ߈ߥᄌൻࠍ઻ࠊߕߦੇ⌁ߣ޿߁഍⊛ߥ↢๮ᵴേ⁁ᘒߩ

77

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ߦㆡߒߡ޿ࠆ‫ߦ⊛⥸৻ޕ‬ਛᔃ὇⚛ઍ⻢ߥߤߩ৻ᰴઍ
⻢ߩਛ㑆ઍ⻢‛⾰ߦߪࠗࠝࡦᕈߩൻว‛߇ᄙߊ‫ߎޔ‬ࠇࠄࠍᱜ⏕ߦಽ㔌࡮᷹ቯߔࠆߚ߼ߦߪ CE-MS ߇ᦨ߽ㆡߒߡ޿ࠆ 21, 28‫ޕ‬ห᭽ߦ‫ޔ‬⢽⾰߿♧ߥߤߩਛᕈ‛⾰ߦߪ LC-MS ߇‫ޔ‬⢽⢌㉄߿ࠕ࡞ࠦ࡯࡞ߥߤߩ᳇ൻ
ߒ߿ߔ޿‛⾰ߦߪ GC-MS ߇ㆡߒߡ߅ࠅ‫ߥ⊛⟜✂ޔ‬ઍ⻢‛⾰ߩ᷹ቯߦߪߤࠇ߆৻ߟߩࠪࠬ࠹ࡓࠍ↪޿ࠆߩߢߪߥߊ‫ߺ⚵ࠍࡓ࠹ࠬࠪߩࠄࠇߎޔ‬วࠊߖߚ⸃ᨆ
߇ਇนᰳߢ޽ࠆ‫࡯ࡠࡏ࠲ࡔߩࠪࡓࡑࠢ࠾ࠝ߫߃଀ޕ‬ࡓ⸃ᨆߦߟ޿ߡߪߎࠇ߹ߢߦ৻ઙߩߺႎ๔߇޽ࠆ߇
1‫⸃ߩߎޔ‬ᨆߢߪ৻ᰴઍ⻢ࠍ⷗ߡ޿ࠆߦ߽㑐ࠊࠄߕಽ㔌⢻߇৻⇟ૐ޿ GC-MS ࠪࠬ࠹ࡓࠍ↪޿ߡ޿ࠆߚ߼‫ޔ‬ઍ⴫⊛ߥ♧߿ࠕࡒࡁ㉄ߥߤߩ㒢ࠄࠇߚ‛⾰ߩߺߒ߆ᬌ಴ߢ߈ߕ✂⟜ᕈ߇ૐߊ‫⚿ޔ‬ᨐߣߒߡੇ⌁ᯏ᭴ࠍ᣿ࠄ߆ߦߔࠆ਄ߢߪᰳ߆ߖߥ޿ోઍ⻢ߩⷰኤߦߪ⥋ߞߡ޿ߥ޿‫ޕ‬ߔࠆߎߣߥߤ߆ࠄ‫ߡ޿߅ߦ⌁ੇߩࠪࡓࡑࠢޔ‬㊀ⷐߥᓎഀࠍᜂߞߡ޿ࠆน⢻ᕈ߇␜ߐࠇߚ‫ޕ‬ታ㓙‫࠸࡛ࠦޔ‬࠽ࠢࡑࡓࠪߦ߅޿ߡߪ CAHS/SAHS ߪㅢᏱᤨߢ߽ᕡ
Ᏹ⊛ߦᄢ㊂⊒⃻ߒߡ߅ࠅ‫ޔ‬ㅦ޿ੇ⌁ࠍⴕ߁↢‛ࠄߒߊᏱߦੇ⌁ߦ஻߃ࠆᚢ⇛߇޽ࠄߚ߼ߡ⏕⹺ߐࠇߚ‫ޕ‬
4. ੇ⌁ߩࡔ࠲ࡏࡠࡒࠢࠬ ࡛ࠦ࠸࠽ࠢࡑࡓࠪߢߪᣂⷙߩ࠲ࡦࡄࠢࡈࠔࡒ࡝࡯
CAHS ߣ SAHS ߇ᕡᏱ⊛ߦᄢ㊂ߦ⊒⃻ߔࠆߎߣߦࠃ
ࠅ‫ޔ‬LEA ߩࠃ߁ߥ࠲ࡦࡄࠢಝ❗ᛥ೙߿ߐ߹ߑ߹ߥੇ
῎ࠬ࠻࡟ࠬᔕ╵ߦߡ௛޿ߡ޿ࠆน⢻ᕈ߇᣿ࠄ߆ߣߥ
ߞߚ߇‫ߡߞࠃߦ῎ੇߪߢߌߛࠇߎޔ‬ᒁ߈⿠ߎߐࠇࠆ
ߪߕߩ㉄ൻࠬ࠻࡟ࠬ߳ߩኻᔕࠍోߡ⺑᣿ߢ߈ࠆࠊߌ
ߢߪߥ޿‫ޔߚ߹ޕ‬CAHS ߿ SAHS ߪᭂ߼ߡᒙ޿ੇ⌁⢻ ജ ߒ߆ ߽ߚ ߥ ޿࠼ ࠘ࠫ ࡖ ࡞࠳ ࡦࡗ ࡑ ࠢࡑ ࡓࠪ(Hypsibius dujardini)ߦ߅޿ߡ߽଻ሽߐࠇߡ޿ࠆߎߣ
߆ࠄ‫ࠆߌ߅ߦࠪࡓࡑࠢ࠽࠸࡛ࠦޔ‬ㅦ޿ੇ⌁ࠍน⢻ߣߔࠆಽሶᯏ᭴ߦߟ޿ߡ߽ߐࠄߥࠆ⸃ᨆ߇ਇนᰳߢ޽
ࠆ‫ߪߢ⌁ੇߩࠪࡓࡑࠢ࠽࠸࡛ࠦޕ‬ォ౮࡮⠡⸶ߦࠃࠆ⊒⃻ᄌേ߇㒢ቯ⊛ߢ޽ࠆߚ߼‫⸃ߣߕ⥄ޔ‬ᨆኻ⽎ߪࠃࠅ⋥ធ⊛ߥ⚦⢩ౝൻว‛ߩᄌൻേᘒ‫ޔ‬හߜ‫ࡏ࠲ࡔޔ‬ࡠ࡯ࡓߣߥࠆ‫ޕ‬ ߘߎߢ‫ޔ‬ᚒ‫ࡦࠝࠗߪޘ‬ᕈൻว‛ߣਛᕈ‛⾰ࠍ✂⟜⊛ߦ⸃ᨆߔࠆߚ߼ߦ‫ޔ‬CE ෸߮ LC ࠪࠬ࠹ࡓߦࠃࠆൻ
ว‛ߩಽ㔌ߣ‫♖ޔ‬ኒ⾰㊂ᢙࠍᓧࠆߚ߼ߦ㘧ⴕᤨ㑆ဳ
(Time-of-flight: TOF)MS ࠍ⚵ߺวࠊߖߚ CE-TOFMS෸߮ LC-TOFMS ߦࠃߞߡ࡛ࠦ࠸࠽ࠢࡑࡓࠪߩઍ⻢ᄌൻࠍ⺞ߴߚ‫⚿ߩߘޕ‬ᨐ‫ޔ‬㧤⒳ߩ♧‫ޔ‬ว⸘ 373 ⒳ߩ
⢽⾰‫ޔ‬269 ⒳ߩหቯߐࠇߚࠗࠝࡦᕈൻว‛‫ߡߒߘޔ‬1328 ⒳ߩᧂ⍮ࡇ࡯ࠢࠍᓧߚ‫ޕ‬ ࡔ ࠲ ࡏ ࡠ ࡯ ࡓ ⸃ ᨆ ߦ ߪ ⾰ ㊂ ಽ ᨆ ེ (Mass
Spectrometer: MS)ࠍ↪޿ࠆ߇‫⚦ޔ‬⢩߆ࠄ᛽಴ߐࠇߚ
ൻว‛ߪ⾰㊂ಽᨆེߦ߆ߌߡߘࠇߙࠇߩ㊂ࠍ᷹ቯߔࠆ೨ߦ୘‫ߦޘ‬ಽ㔌ߐࠇࠆᔅⷐ߇޽ࠆ‫ࡔޔ߼ߚߩߘޕ‬࠲ࡏࡠ࡯ࡓ⸃ᨆߦ߅޿ߡߪᶧ૕ࠢࡠࡑ࠻ࠣ࡜ࡈࠖ
(Liquid Chromatography: LC)‫ࠖࡈ࡜ࠣ࠻ࡑࡠࠢࠬࠟޔ‬(Gas Chromatography: GC)‫࡯࡝࡜ࡇࡖࠠߡߒߘޔ‬㔚᳇ᵒേ(Capillary Electrophoresis: CE)ߥߤߦࠃࠆൻว‛ߩಽ㔌ࠍ⚵ߺวࠊߖߚ LC-MS, GC-MS, CE-MS ߣ޿ߞߚࠪࠬ࠹ࡓࠍ↪޿ࠆ‫ߩࠄࠇߎޕ‬ಽ㔌ᣇᴺߪ᷹ቯኻ⽎ߩൻว‛ߩ⒳㘃ߦࠃߞߡല₸߇⇣ߥࠅ‫ޔ‬LC ߪਛᕈ‛⾰‫ޔ‬GC ߪើ⊒ᕈ‛⾰‫ޔ‬CE ߪࠗࠝࡦᕈ‛⾰ߩಽ㔌 ࠢࡑࡓࠪߪ৻⥸⊛ߦ♧ߦࠃࠆࠟ࡜ࠬൻࠍⴕࠊߥ޿
߇‫߽ߡ޿߅ߦࠪࡓࡑࠢ࠽࠸࡛ࠦޔ‬࿑ 2 ߦ␜ߔࠃ߁ߦ
࠻࡟ࡂࡠ࡯ࠬ߇♧⒳ߩਛߢߪᄙߊሽ࿷ߒߡ߅ࠅ‫߹ޔ‬ߚ‫ߡߞࠃߦ⌁ੇޔ‬ᄢ᏷ߥჇട߇⷗ࠄࠇࠆ‫৻ޕ‬ᣇߢ‫ޔ‬
Ⴧടᓟߩ㊂ߪߎࠇ߹ߢߦᄙߊߩࠢࡑࡓࠪߢႎ๔ߐࠇ
ߡ߈ߚࠃ߁ߦੇ῎㊀㊂ߩ 1%ߦ߽ḩߚߥ޿㊂ߢ޽ࠅ‫ޔ‬⚦⢩ࠍࠟ࡜ࠬൻߒߡ଻⼔ߔࠆߣ޿߁ࠃࠅߪ‫ޔ‬ᶐㅘ࿶ࠬ࠻࡟ࠬߦ߅޿ߡ࠻࡟ࡂࡠ࡯ࠬࠍㇱಽ⊛ߦᵴ↪ߒߡ޿ࠆߣ⷗ࠆߴ߈ߛࠈ߁‫ޕ‬

࿑ 2 㧦࡛ࠦ࠸࠽ࠢࡑࡓࠪㅢᏱᤨ㧔㕍㧕ߣੇ⌁ᤨ㧔⿒㧕ߩ♧Ớᐲ ♧㘃ߩਛߢߪ࠻࡟ࡂࡠ࡯ࠬ߇㗼⪺ߦᄙߊ‫ ⚂ߦᤨ⌁ੇޔ‬3.5 ୚ߦ߹ߢჇടߒߡ޿ࠆ߇‫῎ੇ߽ߢࠇߘޔ‬㊀㊂ߩ
0.735%⒟ᐲߩ㊂ߢ޽ࠅ‫⫾ࠬ࡯ࡠࡂ࡟࠻ޔ‬Ⓧဳߩࡀࡓ࡝࡙ࠬ࡝ࠞߥߤߩੇ⌁ᯏ᭴ߣߪ⇣ߥࠆߎߣ߇ࠊ߆ࠆ‫ޕ‬78

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)࿑ 3 㧦࡛ࠦ࠸࠽ࠢࡑࡓࠪౝࠞ࠴ࠝࡦಽሶߩࡔ࠲ࡏࡠ࡯ࡓ⚿ᨐ‫ޕ‬਄ߩ೉߇ㅢᏱᤨߩ⚿ᨐ‫ޔ‬ਅߩ೉߇ੇ⌁ᤨߩ⚿ᨐ‫ޔ‬Ꮐߩ೉߇ൻว‛ࠍหቯߢ߈ߚࡇ࡯ࠢ‫ޔ‬ฝ߇ᧂหቯߩࡇ࡯ࠢ‫ޕ‬หቯߐࠇߚࡇ࡯ࠢߢߪ GlycerophosphocholineߩჇട߇ tun ߢ㗼⪺ߢ޽ࠅ‫ᧂޔ‬หቯߩࡇ࡯ࠢߢߪ⾰㊂ 170.1049 ߩᧂ⍮‛⾰߇⫾Ⓧߒߡ޿ࠆߎߣ߇ࠊ߆ࠆ‫ޕ‬ฦઍ⻢‛⾰ߪ⾰㊂/㔚⩄(m/z)ߣࠠࡖࡇ࡜࡝࡯㔚᳇ᵒേߩᵒേᤨ㑆㧔MT㧕ߢቯ⟵ߐࠇࠆ‫ޕ‬ሶਛߦᜬߟࡌ࠲ࠗࡦߣߒߡ⚦⢩⾰ਛߩㆆ㔌㊄ዻࠗࠝࡦࠍ✭ⴣߔࠆ௛߈߇ႎ๔ߐࠇߡ޿ࠆ 8‫࡛ޔߦࠄߐޕ‬ࠦ࠸࠽ࠢࡑࡓࠪߩੇ⌁ߢߪઁߦ߽ߐ߹ߑ߹ߥੇ῎ࠬ࠻࡟ࠬᔕ╵߇⿠߈ߡ޿ࠆߎߣ߇߁߆߇߃ࠆ‫ޕ‬GPC ߩઁߦ߽㉄ൻࠬ࠻࡟ࠬᔕ╵ߢઍ⴫⊛ߥ㉄ൻဳࠣ࡞࠲࠴
ࠝࡦ߿ࡆࠝ࠴ࡦ߽ᄢ߈ߊჇടߒߡ޿ࠆઁ‫᦭ޔ‬ᗧߥჇടࠍ␜ߔൻว‛ߩᄙߊߪࡌ࠲ࠗࡦ߆ࠠ࡟࡯࠻૞↪ࠍᜬߟಽሶߢ޽ߞߚ‫ࠄࠇߎޕ‬ઍ⻢ߩో૕௝ࠍ‫◲ޔ‬නߩߚ߼ߦ᦭ᗧߦᄌേߒߚ‛⾰ߩߺࠍขࠅ಴ߒ‫ޔ‬Pathway
Projector ࠍ↪޿ߡ KEGG ࠣࡠ࡯ࡃ࡞ࡑ࠶ࡊ਄ߦนⷞ
ൻߒߚ߽ߩ߇࿑ 4 ߢ޽ࠆ‫ߡߞࠃߦ⌁ੇޕ‬ઍ⻢߇৻ᣤ஗ᱛߔࠆߚ߼‫࡯ࠡ࡞ࡀࠛޔ‬Ḯߢ޽ࠆ ATP ߥߤߩᩭ㉄
߿ࠕࡒࡁ㉄ߩ߶ߣࠎߤߪᕆỗߦ૶޿ዧߊߐࠇᷫዋߔ
ࠆ‫ߦࠇߘޕ‬๭ᔕߒ‫ޔ‬GPC ߿ࠣ࡞࠲࠴ࠝࡦ‫ࡦ࠴ࠝࡆޔ‬ߥߤߣ޿ߞߚ㉄ൻ࡮ᶐㅘ࿶ࠬ࠻࡟ࠬኻᔕߩࡄࠬ࠙ࠚ
ࠗ߇੣ㅴߒ‫ޔ‬ਛ㑆ઍ⻢‛⾰ߪߘࠇߙࠇߩࡄࠬ࠙ࠚࠗ
ߢᦨ߽ㄭறߩࠠ࡟࡯࠻߿ࡌ࠲ࠗࡦ߳ߣઍ⻢࡮วᚑߐࠇ‫⫾ޔ‬Ⓧߐࠇࠆ‫ޕ‬හߜ‫ࠆ߁߈ߢޔ‬㒢ࠅ⍴ᤨ㑆ߢ‫ޔ‬ᱷߐࠇߚࠛࡀ࡞ࠡ࡯ࠍ᦭ലᵴ↪ߒߥ߇ࠄోઍ⻢♽ߢੇ῎ߦࠃࠆᶐㅘ࿶࡮ࠗࠝࡦ࡮㉄ൻߩࠬ࠻࡟ࠬߦኻᔕߒ
ߡ޿ࠆ᭽ሶ߇⷗߃ߡ߈ߚ‫ޕ‬ ߢߪ࡛ࠦ࠸࠽ࠢࡑࡓࠪߩㅦ޿ੇ⌁ࠍታ⃻ߒߡ޿ࠆઍ⻢ᯏ᭴ߪ૗ߥߩߛࠈ߁߆‫߽ᦨޕ‬㗼⪺ߥ‛⾰ߩᄌൻߪࠞ࠴ࠝࡦߦ⷗ࠄࠇߚ㧔࿑ 3㧕‫ޕ‬ㅢᏱᤨߣੇ⌁ᤨߩࡔ࠲ࡏࡠ࡯ࡓࡊࡠࡈࠔࠗ࡞ࠍᲧセߔࠆߣ‫ޔ‬ᄢᄙᢙߩઍ
⻢‛⾰ߦߪᭂ┵ߥᄌേߪ⷗ࠄࠇߥ޿߽ߩߩ‫ޔ‬
Glycerophosphocholine(GPC)߇⪺ߒ޿Ⴧടࠍ⷗ߖߡ
޿ࠆ‫ޕ‬㊂⊛ߦߪ GPC ⒟ߩ⫾Ⓧ߇ߥ޿߇‫ޔ‬ᄌൻ㊂ߢߪㅢᏱᤨߣੇ⌁ᤨߩઍ⻢‛⾰㊂ߦ᦭ᗧߥჇട(Welcht-test, p < 0.05)ࠍ⷗ߖߚ 17 ⒳ߩൻว‛ߩ߁ߜ਄૏㧠⒳ࠍࠦ࡝ࡦઍ⻢♽ߩਛ㑆ൻว‛㧔Phosphorylcholine,
GPC, Ethanolamine phosphate, Choline, ߘࠇߙࠇ⚂ 3
ޯ8 ୚ᄌേ㧕߇භ߼ߡ޿ߚ‫ߦᤨ⌁ੇߪߢࡦࠝ࠾ࠕޕ‬᦭ᗧߦჇടߒߚ‛⾰ 22 ⒳ਛ 7 ⒳ࠍ⇣ߥࠆ㐳ߐߩ⢽⢌㉄߇භ߼ߡ߅ࠅ‫ޔ‬⢽⾰߇ో૕⊛ߦᷫዋߒߡ޿ࠆߎߣ
߆ࠄ‫ޔ‬⢽⾰ࠍಽ⸃ߒߡᓧࠄࠇߚࠣ࡝࠮ࡠ࡯࡞ࠍ૶↪
ߒߡ GPC ࠍ⍴ᤨ㑆ߢวᚑߒߡ޿ࠆ㕖Ᏹߦ࠳ࠗ࠽ࡒ
࠶ࠢߥੇ⌁ᔕ╵߇⿠߈ߡ޿ࠆߣ⠨߃ࠄࠇࠆ‫ޕ‬GPC ߪᏱߦᶐㅘ࿶ࠬ࠻࡟ࠬߦߐࠄߐࠇࠆ⣢⤳ߥߤߢਥⷐߥ
௛߈ࠍߔࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ㉄ൻࠬ࠻࡟ࠬ߿ᶐㅘ
࿶ࠬ࠻࡟ࠬߦ߅޿ߡ㊀ⷐߥಽሶߢ޽ࠅ‫⾰ࠢࡄࡦ࠲ޔ‬ߩ଻⼔߿‫ޔ‬᳓ಽሶߩࠃ߁ߦᱜ㔚⩄ߣ⽶㔚⩄ࠍห৻ಽ79

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)࿑ 4 㧦࡛ࠦ࠸࠽ࠢࡑࡓࠪੇ⌁ߩᲧセࡔ࠲ࡏࡠࡒࠢࠬ
KEGG Pathway ਄ߦ᦭ᗧߥᄌൻࠍ␜ߒߚൻว‛ߩ fold change ࠍนⷞൻߒߚ‫ᤨ⌁ੇޕ‬㧔tun㧕ߦ਄᣹ߒߚ‛⾰߇⿒ߢ‫ޔ‬ᷫዋߒߚ‛⾰߇㕍ߢ␜ߐࠇߡ޿ࠆ‫࡯ࠡ࡞ࡀࠛޕ‬ઍ⻢߇ో૕⊛ߦᷫዋߔࠆਛߢ‫ޔ‬ฦઍ⻢⚻〝ᲤߦჇടߒ
ߡ޿ࠆ‛⾰߇ሽ࿷ߔࠆ‫ޕ‬

࿑ 5 㧦࡛ࠦ࠸࠽ࠢࡑࡓࠪੇ⌁ߦ߅ߌࠆઍ⻢ᔕ╵ߩ߹ߣ߼⢽⾰߇ಽ⸃ߐࠇ‫ߩߘޔ‬ਛ㑆ઍ⻢‛⾰ߢ޽ࠆⶄᢙߩ⢽⢌㉄߇਄᣹ߔࠆ߇‫ߪ࡞࡯ࡠ࠮࡝ࠣޔ‬ਥߦ㉄ൻࠬ࠻࡟ࠬᔕ
╵ߦ㊀ⷐߥ Glycerophosphocholine ߩวᚑߦ૶ࠊࠇࠆ‫߿㉄ࡁࡒࠕ߽ߦઁޕ‬㜞ࠛࡀ࡞ࠡ࡯࡝ࡦ㉄⚿วࠍᜬߟᩭ㉄߿ߥߤ߇ࡌ࠲ࠗࡦ߿ࠠ࡟࡯࠻ಽሶ߳ߣઍ⻢ߐࠇࠆߎߣߦࠃࠅ‫ోࠗࠚ࠙ࠬࡄޔ‬૕ߢേ⊛ߦੇ῎ࠬ࠻࡟ࠬߦኻᔕ
ߒߡ޿ࠆ᭽ሶ߇߁߆߇߃ࠆ‫ޕ‬

80

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)5. ੹ᓟߩዷᦸ
࡛ࠦ࠸࠽ࠢࡑࡓࠪߪઁߩࠢࡑࡓࠪߦᲧߴࠆߣ㘺⢒߇ኈᤃߢ޽ࠆ߇‫ޔ‬Ⴧᱺല₸ߪઁߩࡕ࠺࡞↢‛ߦᲧߴࠆߣᖡߊ‫ޔ‬୘૕ࠨ߽ࠗ࠭ዊߐ޿ߎߣ߆ࠄᄢⷙᮨߥࡔ࠲
ࡏࡠ࡯ࡓ⸃ᨆߥߤࠍⴕ߅߁ߣߔࠆߣචಽߥࠨࡦࡊ࡞
㊂ࠍᓧࠆߩߦᄙᄢߥᤨ㑆ࠍⷐߔࠆ‫⃻ޔ߼ߚߩߘޕ‬Ბ㓏ߢߪ߹ߛㅢᏱ⁁ᘒߣੇ⌁ᤨߩᲧセࡔ࠲ࡏࡠࡒࠢࠬ
ߩߺ߇㆐ᚑߐࠇߡ޿ࠆ‫৻ޕ‬ᣇߢ‫ߩ⌁ੇޔ‬ಽሶᯏ᭴ࠍ
ℂ⸃ߔࠆߚ߼ߦߪੇ⌁߳ߩ⒖ⴕߛߌߢߥߊ‫ޔ‬஗ᱛߐࠇߚઍ⻢ߩౣ㐿ࠍ⸃ᨆߔࠆߎߣ߽ਇนᰳߢ޽ࠅ‫߹ޔ‬ߚ‫♽ᤨࠍࠇߘޔ‬೉ߢⷰኤߔࠆߎߣ߇ᦸ߹ߒ޿‫ޕ‬ታ㓙‫ޔ‬ੇ⌁ᤨߦ⪺ߒ޿⫾Ⓧ߇⷗ࠄࠇߚ GPC ߣࠦ࡝ࡦઍ⻢ߪࠣ࡝࠮ࡠ࡝ࡦ⢽⾰ߩวᚑߣኒធߥ㑐ଥߦ޽ࠅ‫⚦ޔ‬⢩⤑ߩ଻ోߦ߅޿ߡ㊀ⷐߥᓎഀࠍṶߓࠆ‫߅ߦ⌁ੇޕ‬ߌࠆ⣕᳓ߪᅤ૗ߦߐ߹ߑ߹ߥࠬ࠻࡟ࠬᔕ╵ߦࠃߞߡ଻⼔╷߇ߣࠄࠇߡ޿ࠆߣߒߡ߽↢૕⤑ߦዋߥ߆ࠄߧ
៊்ࠍਈ߃ࠆน⢻ᕈߪචಽߦ޽ࠆߣ⠨߃ࠄࠇࠆߩߢ‫ޔ‬ࠬ࠻࡟ࠬᔕ╵ߣߒߡ GPC ࠍ⫾Ⓧߒߟߟੇ⌁ᤨߪ⚦⢩ࠍ଻⼔ߒ‫ޔ‬ๆ᳓ᓟߪߎࠇࠍઍ⻢ߒߡ⤑ߩ଻ోߦᵴ
↪ߒߡ޿ࠆߣ⠨߃ࠇ߫‫ޔ‬㕖Ᏹߦวℂ⊛ߥࠪࠬ࠹ࡓߢ
޽ࠆࠃ߁ߦᕁ߃ࠆ‫੹ޔߡߞࠃޕ‬ᓟઍ⻢ౣ㐿ᤨߩࡔ࠲ࡏࡠࡒࠢࠬߣ‫ߩߘޔ‬ઍ⻢ࠍᜂ߁㉂⚛ߩ೙ᓮߦߟ޿ߡ
߽วࠊߖߡ⸃ᨆߒߡ޿ߊߎߣߢ‫ߩߡߒߣࡓ࠹ࠬࠪޔ‬ੇ⌁ࠍℂ⸃ߒߡ޿ߊߎߣ߇ߢ߈ࠆߩߢߪߥ޿߆ߣᕁ
߁‫ޕ‬⻢ㄉ ᧄ⎇ⓥߪᘮᙥ⟵Ⴖᄢቇవ┵↢๮⑼ቇ⎇ⓥᚲ દ⮮ථᦶ⎇ⓥຬ‫ᦥޔ‬ᚒ᦮⟵ᢎ᝼‫ޔ‬౷↰ൎᢎ᝼‫᧲ޔ‬੩ᄢቇ ࿡ᨑᱞ๺ഥᢎ‫ ╙࡝ࡄޔ‬5 ᄢቇ ၳᎹᄢ᮸⎇ⓥຬߣߩ౒ห⎇ⓥߢ޽ࠆ‫⎇ᧄޔߚ߹ޕ‬ⓥߪ⑼ቇ⎇ⓥ⾌⵬ഥ㊄ ⧯
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Journal of Japanese Society for Extremophiles (2012) Vol.11, 83-89

Abe F.a,b

Direct measurement of membrane properties in piezophiles under high pressureusing fluorescence depolarization techniqueᅢ࿶ᕈ⚦⩶ߦ߅ߌࠆ⤑‛ᕈߩ⋥ធ⸘᷹—㜞࿶Ⱟశ஍శ⸃ᶖᴺ߆ࠄߩࠕࡊࡠ࡯࠴—㕍ጊቇ㒮ᄢቇℂᎿቇㇱ ൻቇ࡮↢๮⑼ቇ⑼㧔⁛㧕ᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ ᶏᵗ࡮ᭂ㒢ⅣႺ↢‛࿤㗔ၞ㒙ㇱ ᢥᔟaDepartment of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin University,
5-10-1 Fuchinobe, Chuo-ku, Sagamihara 252-5258, Japan.
bInstitute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15
Natsushima-cho, Yokosuka 237-0061, Japan
Corresponding author: Fumiyoshi Abe, abef@chem.aoyama.ac.jp
Phone: +81-42-759-6233, Fax: +81-42-759-6511
Received: November 13, 2012 / Accepted: November 23, 2012ߪߓ߼ߦ

Abstract Deep sea is characterized by high pressure
and low temperature, and the organisms inhabiting theenvironment have adapted to such extreme conditions.
High pressure and low temperature exert profoundphysiological impacts on bilayer membranes, primarily
resulting in tighter packing and restricting the rotationalmotion of acyl chains. The maintenance of appropriate
membrane fluidity is crucial for the integrity ofmembrane proteins and cell survival under high pressure
and low temperature. Of the spectroscopic techniques
available to study membrane properties, fluorescenceanisotropy measurement is a common useful method
providing information on dynamic membrane properties.
Recently we developed a new system that enabled
time-resolved fluorescence anisotropy measurement
under high pressure. In this paper, I describe a renewed
view of the membrane properties in deep-sea piezophile
Shewanella violacea when the cells are exposed to highpressure.

 2012 ᐕ 3 ᦬ 26 ᣣ‫ޔ‬ᤋ↹⋙〈ߩࠫࠚ࡯ࡓ࠭࡮ࠠࡖ
ࡔࡠࡦ᳁߇ࡑ࡝ࠕ࠽ᶏḴߩ਎⇇ᦨᷓㇱ 1 ਁࡔ࡯࠻࡞
ߩ࠴ࡖ࡟ࡦࠫࡖ࡯ᶏᷗߦ೔㆐ߒߚ‫ޕ‬න⁛ẜ⥶ߣߒߡߪ਎⇇ೋߢ޽ࠆ‫ߦࠇߘޕ‬వ┙ߟ 1995 ᐕ‫ⴕ┙⁛ޔ‬᡽ᴺ
ੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ߩήੱẜ᳓⦈̌߆޿ߎ߁̍ߪห
ߓ࠴ࡖ࡟ࡦࠫࡖ࡯ᶏᷗߢᐩᵆࠨࡦࡊ࡝ࡦࠣߦᚑഞߒ
ߡ޿ࠆ‫ߩ߁ߎ޿߆ޕ‬តᩏߢಽ㔌ߐࠇߚ 3,000 ߽ߩᓸ↢‛ᩣߪ‫ޔ‬1000 ᳇࿶߽ߩ㜞᳓࿶ߦߐࠄߐࠇߚᷓᶏ߇
ᱫߩ਎⇇ߢߪߥ޿ߎߣࠍ‛⺆ߞߚ‫ޕ‬ᶏߩᐔဋᷓᐲߪ
ᗧᄖߦᷓߊ߅ࠃߘ 3,800 m ߦ㆐ߔࠆ‫ޕ‬࿾⃿ో૕ߩ૕Ⓧ߆ࠄ឵▚ߔࠆߣߎߩ࿾⃿ߩᄢㇱಽߪ㒽਄ߩ 380 ୚‫ޔ‬
ታߦ 380 ᳇࿶߽ߩ㜞࿶ജߦߐࠄߐࠇߚⅣႺߢ޽ࠆ‫ޕ‬ߥ߅‫ޔ‬࿶ജߩ࿖㓙න૏ߪࡄࠬࠞ࡞ (Pa) ߢ޽ࠅ‫ᧄޔ‬Ⓜߢ߽ߎࠇࠍ↪޿ࠆ‫ޕ‬㒽਄ߪ 1 ᳇࿶ߥߩߢ 0.1 MPa‫ޔ‬ᷓߐ 1,000 m ߢߪ 10 MPa‫ޔ‬10,000 m ߢߪ 100 MPa ߽ߩ᳓࿶߇߆߆ࠆ‫ޕ‬‫ߡߒ߁ߤޡ‬ᷓᶏ↢‛ߪߎࠇ߶ߤߩ⿥
㜞࿶ജਅߢ↢߈ࠄࠇࠆߩ߆㧫‫࡯ࡖࠫࡦ࡟ࡖ࠴ޢ‬ภߩ⥶ᶏ߆ࠄ 140 ᐕ‫⺰ߩߚ߹޽ޔ‬ᢥࠍ߽߭⸃޿ߡߺߡ߽
᳿ቯ⊛ߥ╵߃ߪ⷗ߟ߆ߞߡ޿ߥ޿‫ޔߒ߆ߒޕ‬ේᆎ↢๮⺀↢ߩ႐߇ᷓᶏߛߞߚߣߔࠆߥࠄ߫‫ޔ‬‫ߡߞ߿߁ߤޡ‬㒽਄↢‛ߪߎࠇ߶ߤߩૐ࿶ജਅߦㆡᔕߒㅴൻߒߡ߈
ߚߩ߆㧫‫ߚߞ޿ߣޢ‬໧޿߆ߌߦ⟎߈឵߃ࠄࠇࠆߛࠈ
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ࠄࠇࠆὐ߇⥝๧ᷓ޿‫ޕ‬ᅢ࿶ᕈ⚦⩶ߦߟ޿ߡߪ‫࡝ࠞޔ‬ࡈࠜ࡞࠾ࠕᄢቇࠬࠢ࡝࠶ࡊࠬᶏᵗ⎇ⓥᚲߩ D. H.
Bartlett ࠄߩ Photobacterium profundum SS9 ߣᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ߩട⮮ࠄߩ Shewanella violacea DSS12 ࠍᓸ↢‛ࡕ࠺࡞ߣߒߚಽሶ↢‛ቇ⊛⎇ⓥ߇♖ജ⊛ߦㅴ߼ࠄࠇߡ޿ࠆ‫ߩࠄߜߤޕ‬ᅢ࿶ᕈ⚦⩶߽ㆮવሶ⊒⃻߿࠲ࡦࡄࠢ⾰ᯏ⢻ߦ㑐ߔࠆᄙᢙߩႎ๔߇޽ࠅ‫ᦨޔ‬ㄭߪోࠥࡁࡓ㈩೉߽⸃⺒ߐࠇ‫ߩࠄࠇߘ߽ߢ⺕߿੹ޔ‬ㆮવሶࠍࠢࡠ࡯࠾ࡦࠣߢ߈ࠆࠃ߁ߦߥߞߚ 4, 21)‫ޕ‬߁‫ޕ‬ᷓᶏ↢‛ߩ↢ᘒߪ⻘ߦ൮߹ࠇߡ߅ࠅ‫߽੹ޔ‬ᄙߊߩ⎇ⓥ⠪ߩ⥝๧ࠍᗖ߈ߟߌߡ޿ࠆ‫ޕ‬ ZoBell ߣ Johnson ߪ”barophile”ߣ޿߁ᓸ↢‛⒳ࠍ
ቯ⟵ߒߚ‫ޕ‬Barophile ߪ㜞࿶ജࠍᅢࠎߢ↢⢒ߔࠆᓸ↢‛̌ᅢ࿶ᕈ⚦⩶̍ࠍ␜ߔ‫ޕ‬ZoBell ߣ Morita ߪᷓᶏᐩᵆ⹜ᢱ߆ࠄ㜞࿶ജਅߢჇᱺߔࠆᓸ↢‛ߩሽ࿷ࠍೋ߼
ߡ⏕⹺ߒ‫ߩߘޔ‬ᓟ 1979 ᐕߦ Yayanos ࠄ߇ᅢ࿶ᕈ⚦⩶ߩන㔌ߦᚑഞߒߚ 23)‫ޕ‬Yayanos ߪߩߜߦ‫”ޔ‬barophile”
ࠍ”piezophile”ߣ๭߱ߎߣࠍឭ໒ߒ‫⃻ޔ‬࿷ᐢߊฃߌ౉
ࠇࠄࠇߡ޿ࠆ‫ޕ‬baro ߪ㊀ജߣ޿߁ᗧ๧ว޿߇ᒝ޿ߚ
߼‫ޔ‬piezo ߩᣇ߇࿶ജࠍ⴫ߔߩߦ߰ߐࠊߒ޿‫ޕ‬Yayanos߇ቇળߢߎ߁⹤ߒߡߊࠇߚߎߣࠍⷡ߃ߡ޿ࠆ‫ޕ‬Ⴧᱺ․ᕈࠍ࿶ജߩⷰὐ߆ࠄಽ㘃ߔࠆߣ‫ޔ‬piezophilic㧔ᅢ࿶ᕈ㧕ߩઁߦ piezotolerant㧔⠴࿶ᕈ㧕ߣ piezosensitive㧔࿶ജᗵฃᕈ㧕߇޽ࠆ‫ޕ‬ᄢ᳇࿶ਅߦ᫮ᕷߔࠆ↢‛ߪ‫ޔ‬ၮᧄ⊛ߦ࿶ജᗵฃᕈࠍ␜ߒ‫ޔ‬ട࿶ߔࠆߣჇᱺㅦᐲ߇
ૐਅߔࠆ‫ޕ‬ᅢ࿶ᕈߣߪᄢ᳇࿶ਅࠃࠅ߽㜞࿶ജਅߢჇ
ᱺㅦᐲ߇㜞߹ࠆᕈ⾰ߩߎߣߢ޽ࠆ‫ޔߚ߹ޕ‬⠴࿶ᕈߣߪᄢ᳇࿶ߣ㜞࿶ജਅߢห⒟ᐲߦჇᱺߔࠆ႐วߦ↪޿ࠄࠇࠆ‫ࠍࠇߎޕ‬ᮨᑼ⊛ߦ␜ߒߚߩ߇࿑ 1 ߢ޽ࠆ‫ޕ‬⚦⢩ߣ↢૕ಽሶߦኻߔࠆ࿶ജലᨐ ⴫㧝ߦ⚦⢩߿↢૕ಽሶᯏ⢻ߦᓇ㗀ࠍ߅ࠃ߷ߔ࿶ജ
ߩ߅ࠃߘߩਅ㒢୯ࠍ␜ߔ‫⋡߽ࠇߕ޿ߒߛߚޕ‬቟⒟ᐲ
ߦ⠨߃ߡ޿ߚߛ߈ߚ޿‫ޕ‬50 MPa એਅߢߪ‫⚦ޔ‬⢩ㆇേ߿⚦⢩ಽⵚ‫⤑ޔ‬ャㅍ૕ࠍ੺ߒߚᩕ㙃Ḯขࠅㄟߺߥߤ
ߦ࿶ജߩᓇ㗀߇⃻ࠇࠆ 13)‫ߩ߳⾰ࠢࡄࡦ࠲⤑ޕ‬ᓇ㗀߇ᄢ߈޿ߩߪ‫ޔ‬⢽⾰ੑ㊀ጀߩ࿶❗ߦ઻߁㜞ᰴ᭴ㅧߩᄌ
ൻ߇ᄢ߈޿ߚ߼ߢ޽ࠆ 1, 9, 15)‫ޕ‬50~100 MPa ߢߪォ౮߿⠡⸶‫ޔ‬DNA ⶄ⵾ߥߤߩ㒖ኂ߇㗼⪺ߦߥࠅ‫࠴ࠢࠕޔ‬ࡦ߿࠴ࡘ࡯ࡉ࡝ࡦߩ⣕㊀ว‫࠶࠾࡙ࡉࠨࡓ࡯࠰ࡏ࡝ޔ‬࠻ߩ⸃㔌ߥߤ‫ߩ⾰ࠢࡄࡦ࠲࡯ࡑࠧ࡝ࠝޔ‬፣უ߇ᆎ߹
ࠆ 17–19, 22)‫ߪࠇߎޕ‬⣕㊀ว߿⸃㔌ߦ઻߁⽶ߩ૕Ⓧᄌൻߦࠃࠆ߽ߩߢ‫ޔ‬㔺಴ߒߚ࠲ࡦࡄࠢ⾰⴫㕙ߩ᳓๺ߦࠃ
ࠆነਈ߇ᄢ߈޿‫ߢߎߎޕ‬૕Ⓧᄌൻߣߪᐔⴧ߿෻ᔕߦ
઻߁♽ో૕ߩ૕ⓍᏅࠍᜰߔ‫ࡦ࡝ࡉ࡯ࡘ࠴߫߃଀ޕ‬᳓ṁᶧࠍ⠨߃ߡߺࠆߣ‫ޔ‬㊀วߒߡ޿ࠆߣ߈ߣᲧߴ⣕㊀วߒߡ޿ࠆߣ߈ߩᣇ߇࠴ࡘ࡯ࡉ࡝ࡦ 1 ࡕ࡞޽ߚࠅߩ
૕Ⓧ߇⚂ 90 mL ዊߐ޿ 18)‫ߩࠛ࡝࠻ࡖࠪ࡮࡞ߡߞࠃޕ‬ᴺೣߦᓥ޿‫ޔ‬ട࿶ߔࠆߣ⣕㊀วᣇะߦᐔⴧߪࠪࡈ࠻
ߔࠆ‫ޕ‬100 MPa એਅߩ࿶ജߪ৻⥸ߦ㕖⥌ᱫ⊛ߣ⸒ࠊࠇ‫ޔ‬ᢙಽ‫⚦ޔ‬⢩ߦࠃߞߡߪᢙᤨ㑆ߩട࿶ߢ߽↢ሽ₸߇ᄢ߈ߊૐਅߔࠆߎߣߪߥ޿‫ޔࠈߎߣߩ੹ߒ߆ߒޕ‬ታ㛎ቶ᧦ઙਅߢ 100 MPa ߢߩ⚦⢩ಽⵚ㧔Ⴧᱺ㧕߇⏕⹺ߐࠇߚ଀ߪ‫ޔ‬ᷓᶏ↱᧪ߩᅢ࿶ᕈ⚦⩶ߣ޿߃ߤ߽ႎ
๔ߐࠇߡ޿ߥ޿‫৻ޕ‬ᣇ‫ߩ⾰ࠢࡄࡦ࠲࡯ࡑࡁࡕޔ‬ᄬᵴ߿࿶ജᄌᕈߪ 100~200 MPa ߢᆎ߹ࠅ‫ޔ‬DNA ߿ RNAߦ⥋ߞߡߪ 1 GPa ߹ߢ᭴ㅧ߇቟ቯߢ޽ࠆ 5, 8, 14, 16)‫ޕ‬ᓥߞߡ‫ޔ‬ォ౮߿⠡⸶‫ޔ‬DNA ⶄ⵾ߦ߅ߌࠆ࿶ജ㒖ኂߪᩭ㉄ߘߩ߽ߩߩᄌᕈߢߪߥߊ‫ࠍࠄࠇߘࠈߒ߻ޔ‬ᠲࠆ
DNA/RNA ࡐ࡝ࡔ࡜࡯࠯ⶄว૕߿࡝ࡏ࠰࡯ࡓ߳ߩᓇ
㗀߇ⷐ࿃ߢ޽ࠆ‫ޕ‬S. violacea ߩ RNA ࡐ࡝ࡔ࡜࡯࠯ߦ
ߪ⠴࿶ᕈ߇޽ࠅ‫ޔ‬㜞࿶㔚᳇ᵒേᴺߦࠃࠆ in vitro ⸃ᨆ߆ࠄ 140 MPa ߹ߢߪࠨࡉ࡙࠾࠶࠻᭴ㅧࠍ⛽ᜬߔࠆߎߣ߇ࠊ߆ߞߡ޿ࠆ 12)‫৻ޕ‬ᣇ‫ޔ‬ห৻᧦ઙਅߢᄢ⣺⩶ߩRNA ࡐ࡝ࡔ࡜࡯࠯ߪ⸃㔌ߒߡߒ߹߁‫ޕ‬S. violacea ߩ↢⢒਄㒢࿶ജ߇⚂ 70 MPa ߢ޽ࠆߎߣࠍ⠨߃ࠆߣ‫ޔ‬RNA ࡐ࡝ࡔ࡜࡯࠯ߪታߦ 2 ୚ߩ቟ో₸ࠍ⏕଻ߒߡ޿ࠆߩߛ‫ޕ‬

࿑ 1. ࿶ജゲߢ⷗ߚᓸ↢‛ߩჇᱺ․ᕈ
ߢߪᷓᶏ߆ࠄಽ㔌ߐࠇߚᓸ↢‛ߩోߡ߇ᅢ࿶ᕈ߿⠴࿶ᕈࠍ␜ߔ߆ߣ⸒ߞߚࠄ‫ޔ‬᳿ߒߡߘࠎߥߎߣߪߥ޿‫ޕ‬ㅒߦ㒽਄↱᧪ߩᓸ↢‛ߪߺߥ࿶ജࠍ߆ߌࠆߣჇᱺߢ
߈ߥߊߥࠆ߆ߣ޿߁ߣ‫߽߆ߒޕ޿ߥ߽ߣߎߥࠎߘޔ‬ߎ߁ߒߚჇᱺ․ᕈߪ‫ޔ‬᷷ᐲ߿ၭ࿾⚵ᚑࠍᄌ߃ࠆߣᄢ
߈ߊᄌൻߔࠆ‫ߢ߹ߊ޽ޕ‬ታ㛎ቶߢⷰኤߐࠇߚ৻ߟߩ
ᜰᮡߥߩߢ޽ࠆ‫ࠄ߇ߥߒ߆ߒޕ‬ಽሶߩ⋡ߢᅢ࿶ᕈ⚦⩶ߣ࿶ജᗵฃᕈ⚦⩶ࠍᲧセߔࠆߣ‫ޔ‬ォ౮߿⠡⸶‫࠲ޔ‬ࡦࡄࠢ⾰ߩ᭴ㅧߣᯏ⢻‫ߪ޿ࠆ޽ޔ‬⢽⾰ߩേ⊛᭴ㅧߣ޿ߞߚ᭽‫ߢ࡞ࡌ࡟ߥޘ‬ਔ⠪ߩ࿶ജଐሽᕈߦ㆑޿߇⷗

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)↢૕⤑ߦ߅ߌࠆ㜞࿶ജㆡᔕ ೨㗄ߢߪ࠲ࡦࡄࠢ⾰ࠍਛᔃߣߒߚ࿶ജലᨐࠍㅀߴ
ߚ‫ޔߢߎߘޕ‬ᰴߦ↢૕⤑߳ߩᓇ㗀ߦߟ޿ߡ⠨߃ߡߺߚ޿‫ޕ‬ၮᧄ⊛૞↪ߣߒߡ㜞࿶ജߪ⢽⾰ੑ㊀ጀࠍ࿶❗
ߒ‫࡞ࠪࠕޔ‬㎮ߩ࿁ォࡉ࡜࠙ࡦㆇേࠍ೙㒢ߔࠆ‫ࠪࠕޕ‬࡞㎮ߩ㐳ߐ߿ਇ㘻๺ᐲ‫ޔ‬ᭂᕈ㗡ㇱߩ⒳㘃ߦࠃࠆᏅߪ
޽ࠆ߇‫ੱߨ߻߅߅ޔ‬Ꮏ⢽⾰ੑ㊀ጀߩ⋧ォ⒖᷷ᐲߪ
100 MPa ߢ 20~30͠਄᣹ߔࠆ‫࠻ࡒ࡞ࡄࠫޔ߫߃଀ޕ‬ࠗ࡞ࡈࠜࠬࡈࠔ࠴ࠫ࡞ࠦ࡝ࡦߩ႐ว‫ޔ‬ᄢ᳇࿶ਅߩਥ
ォ⒖᷷ᐲߪ 42͠ߛ߇‫ޔ‬100 MPa ߛߣ 64͠߹ߢ਄᣹ߔ
ࠆ 9)‫↢ޕ‬૕⤑ߪ⚵ᚑ߇ⶄ㔀ߥߩߢ᣿⏕ߥ⋧ォ⒖ߪⷰ
ኤߐࠇߥ޿‫ߩ⾰ࠢࡄࡦ࠲⤑ޔߒ߆ߒޕ‬ᯏ⢻߇㜞࿶ജߢᄢ߈ߊ៊ߥࠊࠇࠆߩߪ‫ߩ⤑ޔ‬࿶❗ߦ઻߁࠲ࡦࡄࠢ
⾰ߩ᭴ㅧᄌൻߦࠃࠆߚ߼ߢ޽ࠆ‫ޕ‬ㆡಾߥ⤑ߩ⁁ᘒࠍ଻ߟߎߣ߇㜞࿶ജਅߩ↢๮⛽ᜬߦ㊀ⷐߢ޽ࠆߎߣߪ‫ޔ‬↢‛ߩૐ᷷ㆡᔕߣૃߡ޿ࠆ߆߽ߒࠇߥ޿‫ޕ‬ૐ᷷߳ߩㆡᔕᚢ⇛ߩ৻ߟߪ⢽⾰ࠕࠪ࡞㎮ߩਇ㘻๺ൻߢ޽ࠆ‫ޕ‬৻ଔ޽ࠆ޿ߪᄙଔਇ㘻๺⢽⢌㉄ߪ┙૕⊛ߦ߆ߐ㜞޿ߚ߼‫ߩ⤑ޔ‬లႯࠍᅹߍ㜞޿ᵹേᕈ߇⛽ᜬߐࠇࠆ‫ߎޕ‬ࠇߦࠃࠅ⤑࠲ࡦࡄࠢ⾰ߩᯏ⢻߇଻ߚࠇࠆߣ޿߁઀⚵ߺߢ޽ࠆ‫ޕ‬Macdonald ࠄߪᵻᶏ߆ࠄᷓᶏ߹ߢ᫮ᕷߔ
ࠆㄭ✼㝼㘃ߩ⚦⢩⤑ࠍ⺞ߴ‫ޔ‬࿶ജߩᓇ㗀ࠍႎ๔ߒߚ‫ޕ‬ᓐࠄߪ Ⱟశ஍శ⹜⮎ 1,6-diphenyl-1,3,5-hexatrine(DPH)ࠍ↪޿‫ޔ‬㜞࿶ജਅߢⰯశ஍శ⸃ᶖ᷹ቯࠍⴕߞ
ߚ‫⚿ߩߘޕ‬ᨐ‫ޔ‬หߓ࿶ജߩ߽ߣߢߪᷓᶏᕈߩ㝼㘃ߩᣇ߇⤑ߩ⒎ᐨ߇ૐߊ‫ޔ‬ᵹേᕈ߇㜞޿࡟ࡌ࡞ߢ⛽ᜬߐࠇߡ޿ࠆߎߣ߇ࠊ߆ߞߚ‫ޔߚ߹ޕ‬᝝₪ߐࠇߚᷓᐲߦ
⋧ᒰߔࠆ࿶ജࠍ߆ߌࠆߣ‫ߩߤޔ‬㝼ߢ߽⤑ߩ⁁ᘒ߇߶߷หߓߦߥࠆߎߣ߇␜ߐࠇߚ 6)‫߇‛↢ޕ‬ⅣႺ᧦ઙߦวࠊߖߡ⤑ߩᵹേᕈࠍ଻ߟߎߩࠃ߁ߥ઀⚵ߺ
85ࠍ‫”ޔ‬Homeoviscous adaptation”ߣ๭߱‫ޕ‬ ᷓᶏᕈ⚦⩶ߩᄙߊߪࠛࠗࠦࠨࡍࡦ࠲ࠛࡦ㉄
㧔eicosapentaenoic acid, એਅ EPA㧕߿࠼ࠦࠨࠛࠠࠨ
ࠛࡦ㉄㧔docosahexaenoic acid, એਅ DHA㧕ߥߤߩᄙଔਇ㘻๺⢽⢌㉄ࠍᄙߊ฽߻ߩߢ‫ߩ⤑ޔ‬ᵹേᕈࠍ㜞߼ࠆߣ⠨߃ࠄࠇߡ޿ࠆ‫ޔߒ߆ߒޕ‬P. profundum SS9 ߢߪ
EPA ᰳ៊ᩣߢ߽㊁↢ᩣߣห᭽ߦ㜞࿶ജਅߢჇᱺߔࠆ‫ޕ‬ߣߎࠈ߇‫৻ޔ‬ଔਇ㘻๺⢽⢌㉄ߩ cis-vaccenic acid
(C18:1)ࠍᰳߊߣ࿶ജᗵฃᕈࠍ␜ߔߩߢ޽ࠆ 3)‫ޕ‬ධᭂߩ ᳖ ਛ ߆ ࠄ ⊒ ⷗ ߐ ࠇ ߚ ૐ ᷷ ⩶ Shewanella
livingstonensis Ac10 ߢߪ‫ޔ‬EPA ߪૐ᷷᧦ઙਅߦ߅ߌ
ࠆ⚦⢩ಽⵚߦਇนᰳߛ߇‫ࡦ࡟ࡇޔ‬ಽሶߩ᜛ᢔㅦᐲߢ
⷗ࠆ㒢ࠅ EPA ߩ᦭ήߪ⤑ߩᵹേᕈߦᓇ㗀߇ߥ޿ߣ⚿⺰ߠߌࠄࠇߡ޿ࠆ 10)‫ޔߚ߹ޕ‬S. violacea DSS12 ߢߪ EPA ࠍᰳ៊ߔࠆߣ㜞࿶ജਅߢჇᱺㅦᐲ߇ૐਅߔࠆ 11)‫ߦ߁ࠃߩߎޕ‬㜞࿶ജਅ߿ૐ᷷ਅߢ EPA ߇ᨐߚߔᓎഀߣ㊀ⷐᕈߦߟ޿ߡߪ‫⩶ޔ‬⒳߿ၭ㙃᧦ઙ‫ߔ⋡⌕ޔ‬ࠆᒻ⾰ߦࠃߞߡ⚿⺰߇⇣ߥࠅ‫ߪ⸃⷗ߥ⊛৻⛔ޔ‬ᓧࠄࠇߡ޿ߥ޿‫߽ߘ߽ߘޕ‬ᓸ↢‛ߩ⚦⢩⤑߇↢߈ߚ߹߹㜞࿶ജਅߢ⸘᷹ߐࠇߚ଀ߪߎࠇ߹ߢߥ߆ߞߚ‫ߎߘޕ‬ߢએ㒠ߢߪ‫ޔ‬㜞࿶ജਅߦ߅ߌࠆᅢ࿶ᕈ⚦⩶ S. violacea
DSS12 ߩ⚦⢩⤑ߩ‛ᕈߣ EPA ߩᓎഀߦߟ޿ߡ‫ߚ⑳ޔ‬ߜߩᦨㄭߩ⍮⷗ߦၮߠ޿ߡ⸃⺑ߔࠆ‫ޕ‬㜞࿶ജਅߦ߅ߌࠆⰯశ஍శ⸃ᶖ᷹ቯ ೨㗄ߢ߰ࠇߚⰯశ஍శ⹜⮎ DPH ߿ߘߩ⺃ዉ૕
1-[4-(trimethylamino)pheny]-6-phenyl-1,3,5-hexatriene(TMA-DPH) ߪ‫ੱޔ‬Ꮏ⤑ߩ‛ᕈࠍ⺞ߴࠆߚ߼ߎࠇ߹
ߢᐢߊ↪޿ࠄࠇߡ߈ߚ‫ޕ‬DPH ߇⢽⾰ੑ㊀ጀߩࠕࠪ࡞
㎮የㇱ㧔⤑ਛᄩㇱ㧕ߦ૏⟎ߔࠆߩߦኻߒ‫ޔ‬TMA-DPH

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ߪᧃ┵ߦᱜ㔚⩄ࠍᜬߟߚ߼⢽⾰ੑ㊀ጀߩᭂᕈ㗡ㇱߦࠕࡦࠞ࡯ߐࠇࠆ‫Ⱟޕ‬శ⇣ᣇᕈ (fluorescence anisotropy,r) ߪᑼ [1] ߢቯ⟵ߐࠇ‫߇⤑ޔ‬೰⋥ߢ޽ࠇ߫ r ୯ߪ㜞ߊ‫ੂޔ‬㔀ߐ߇Ⴧߔߣߘߩ୯ߪૐਅߔࠆ‫ޕ‬r = (IVV – G IVH)/(IVV + 2G IVH)
[1]ߎߎߢ‫ޔ‬I ߪⰯశᒝᐲ‫ޔ‬H ߣ V ߪബ⿠஍శߦኻߒߘ
ࠇߙࠇ᳓ᐔ߅ࠃ߮ု⋥ᣇะߩⰯశᚑಽࠍᗧ๧ߔࠆ‫ޕ‬G ߪⵝ⟎㑐ᢙߢ޽ࠆ‫Ⱟߪߜߚ⑳ޕ‬శኼ๮ߩᤨ㑆ಽ⸃᷹ቯࠍ೑↪ߒ஍శ⸃ᶖᴺࠍⴕߞߡ߅ࠅ㧔એਅ‫ᤨޔ‬㑆ಽ⸃Ⱟశ஍శ⸃ᶖᴺ㧕‫ޔ‬㉂Უߩ⚦⢩⤑ࠛ࡞ࠧࠬ࠹ࡠ࡯
࡞ߣߘߩ⺃ዉ૕߇⤑ߦ߅ࠃ߷ߔᓇ㗀ࠍ⺞ߴߡ߈ߚ 2)‫ޕ‬ᤨ㑆ಽ⸃⢻ߪ߅ࠃߘ 100 ps ߢ޽ࠆ‫ޕ‬r ߩᤨ㑆ᄌൻߪᑼ [2] ߢ⴫ߐࠇࠆ‫ޕ‬r(t) = [IVV(t) – G IVH(t)]/[IVV(t) + 2G IVH(t)][2]⢽⾰ੑ㊀ጀߦၒ߼ㄟ߹ࠇߚಽሶߪ࿁ォㆇേ߇೙㒢ߐࠇߡ޿ࠆߩߢ‫⇣ޔ‬ᣇᕈߩᤨ㑆ᄌൻߪᦨ߽න⚐ߥ৻ᚑಽㄭૃߢᑼ [3] ߢ⴫ߐࠇࠆ‫ޕ‬r(t) = (r0 г r) ̒exp(гt/T) + r
[3]ߎߎߢ‫ޔ‬r0 ߪⰯశಽሶ߇േ޿ߡ޿ߥ޿ߣ߈ߩ⇣ᣇᕈ‫ޔ‬rߪቢోߦ஍శ⸃ᶖߒߚߣ߈ߩ⇣ᣇᕈ‫ޔ‬ǰߪⰯశಽሶߩ࿁ォ⋧㑐ᤨ㑆ߢ޽ࠆ‫ޕ‬㊀ⷐߣߥࠆߩ߇⒎ᐨ࿃ሶ
(S, order parameter)ߣⰯశಽሶߩ࿁ォ⋧㑐ଥᢙ (Dw,rotational diffusion coefficient) ߢ޽ࠅ‫ࠇߙࠇߘޔ‬ᑼ[4]ߣ[5]ߢ⴫ߐࠇࠆ‫ޕ‬S = (r /r0)1/2[4]
Dw = (r0 г r)/6Tr0[5]⤑ߩ⒎ᐨ߇ૐਅߔࠆߣ S ߪૐਅߒ‫࡞ࠪࠕޔ‬㎮ߩ࿁ォㆇേ߇ỗߒߊߥࠆߣ Dw ߪ㜞ߊߥࠆ‫ᦨޕ‬ㄭ‫ߜߚ⑳ޔ‬
ߪ 200 MPa ߹ߢߩ㜞࿶ജਅߢᤨ㑆ಽ⸃Ⱟశ஍శ⸃ᶖ
ᴺࠍⴕ߁ࠪࠬ࠹ࡓࠍ㐿⊒ߒ‫ ࠍࠇߎޔ‬High-pressure
time-resolved fluorescence anisotropy measurement
system (HP-TRFAM) ߣ๭ࠎߢ޿ࠆ 20)‫ޕ‬࿑ 2. ߦߘߩࠪࠬ࠹ࡓ᭴ᚑࠍ␜ߔ‫ߢߎߘޕ‬ᰴߦ‫ޔ‬HP-TRFAM ࠍ↪
޿ߡ S. violacea DSS12 ߩ⤑‛ᕈࠍ⸃ᨆߒߚ⚿ᨐߦߟ޿ߡ⚫੺ߔࠆ‫ޕ‬

࿑ 2. 㜞࿶ᤨ㑆ಽ⸃Ⱟశ஍శ⸃ᶖ᷹ቯࠪࠬ࠹ࡓ HP-TRFAM
߽ 150 MPa ߹ߢട࿶ߒߡ߽ߎߩ୯ߪᄢ߈ߊᄌൻߖߕ‫ޔ‬㊁↢ᩣߩ⚦⢩⤑ߪᏱߦ೰⋥ߢ޽ࠆࠃ߁ߛ 20)‫ޕ‬EPA ࠍᄬ߁ߣ⤑ߪ೰⋥ߦߥࠆߩ߆ߣ੍ᗐߒߡ޿ߚ߇‫ޔ‬ㅒߦ'pfaA ᩣߩ S ߪ㊁↢ᩣࠃࠅ߽߻ߒࠈૐ޿୯ࠍ␜ߒߚ‫ޕ‬ߟ߹ࠅ EPA ߇޽ࠆᣇ߇⤑ߪ࿕޿ߩߢ޽ࠆ‫(ޕ‬࿑ 4A)‫ޕ‬ߎࠇࠍᡰᜬߔࠆࠃ߁ߦ‫ޔ‬Dw ߪ'pfaA ᩣߢ㜞ߊࠕࠪ࡞㎮ߩ࿁ォㆇേ߇㜞߹ߞߡ޿ߚ (࿑ 4B)‫ޕ‬ ߢߪ‫ ߗߥޔ‬EPA ࠍ฽߻㊁↢ᩣߩ⚦⢩⤑ߪ‫ޔ‬EPA ࠍ฽߹ߥ޿'pfaA ᩣࠃࠅ೰⋥ߥߩߛࠈ߁㧫ታߪ‫ޔ‬㊁↢

ᅢ࿶ᕈ⚦⩶ S. violacea DSS12 ߩ⚦⢩⤑ߣ EPA ߩᓎഀ S. violacea DSS12 ߩ㊁↢ᩣߪ 50 MPa ߢ⦟ᅢߦჇᱺ
ߔࠆ߇‫'ޔ‬pfaA ᩣߢߪჇᱺㅦᐲ߇ૐਅߔࠆ (࿑ 3) 20)‫ޕ‬
TMA-DPH ߢ࡜ࡌ࡞ߒߚ↢⚦⢩ࠍ 10 ͠ߢ 0.1~150
MPa ߩ࿶ജਅߢ HP-TRFAM ࠍⴕߞߚ‫⚿ߩߘޕ‬ᨐ‫ޔ‬㊁↢ᩣߩ⚦⢩⤑ߪᄢ᳇࿶ਅߢ߽㕖Ᏹߦ೰⋥ߢ S ߪ
0.9 ߣ޿߁㜞޿୯ࠍ␜ߔߎߣ߇ࠊ߆ߞߚ (࿑ 4A)‫ޕ‬ᄢ⣺⩶ߥߤߣᲧߴߡߪࠆ߆ߦ⤑ߩᵹേᕈߪૐ޿‫߆ߒޕ‬86

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)᭴ㅧࠍߣࠅ‫ߩ⤑ޔ‬೰⋥ᕈࠍ㜞߼ߡ޿ࠆߩ߆߽ߒࠇߥ
޿‫৻ޕ‬ᣇ‫ޔ‬୚Ⴧߒߚࡄ࡞ࡒ࠻࡟ࠗࡦ㉄ߪ'pfaA ᩣߩ⤑ࠍ߆ߐ㜞ߊߒ‫ޔ‬೰⋥ᕈߩૐਅࠍᒁ߈⿠ߎߒߚߣ⸃
㉼ߐࠇࠆ‫'ޔߒߛߚޕ‬pfaA ᩣߢ⤑ߩ೰⋥ᕈ߇ૐਅߒ
ߚߣߪ޿߃‫ޔ‬ଐὼ‫ޔ‬37 ͠ߦ⥋ㆡჇᱺ᷷ᐲࠍᜬߟࠃ߁
ߥᄢ⣺⩶ߣᲧߴࠆߣ'pfaA ⤑ߪ࿶ୟ⊛ߦ࿕޿‫ޕ‬ᓥߞߡ‫ࠆ޽ޔ‬ᓸ↢‛ߦ߅޿ߡ⢽⾰⚵ᚑ߇ዋߥ߆ࠄߕᄌൻ
ߒߡ߽‫․ޔ‬ቯߩ▸࿐ౝߢ⤑‛ᕈ߇⛽ᜬߐࠇߡ޿ࠇ߫
ᱫߥߕߦ↢߈ߡ޿ࠄࠇࠆ‫ޕ߁ࠈ޽߽㉼⸃ߚߞ޿ߣޔ‬ᩣߩో⢽⢌㉄ߦභ߼ࠆ EPA ߩഀวߪ⚂ 11㧑ߥߩߛ߇‫'ޔ‬pfaA ᩣߢߪ EPA ߇ᶖᄬߔࠆߣߣ߽ߦ‫ࡒ࡞ࡄޔ‬࠻࡟ࠗࡦ㉄㧔palmitoleic acid, C16:1㧕ߩഀว߇⚂ 24%
߆ࠄ 49%ߦჇᄢߒߡ޿ߚ‫ޔߜࠊߥߔޕ‬EPA ߩᰳ៊ߦ઻޿⚦⢩⤑⢽⾰ࠍ᭴ᚑߔࠆ⢽⢌㉄߇ᄢⷙᮨߦౣ᭴▽ߐࠇߡ޿ߚߩߢ޽ࠆ 20)‫ޕ‬ಽሶࠪࡒࡘ࡟࡯࡚ࠪࡦߦࠃ
ࠆߣ‫ޔ‬DHA㧔C22:6㧕ߪ⤑ౝߢࠦࠗ࡞⁁ߣߥࠅᐔဋ
㐳߇ 0.82 nm ߥߩߦኻߒ‫㉄ࡦࠗ࡟ࠝޔ‬㧔oleic acid,
C18:1㧕ߢߪ 1.42 nm ߣႎ๔ߐࠇߡ޿ࠆ 7)‫ޕ‬EPA ߽⢽
⾰ੑ㊀ጀߩ⴫ጀઃㄭߢࠦࠗ࡞ߩࠃ߁ߦࠦࡦࡄࠢ࠻ߥ

࿑ 3. S. violacea ߩ㊁↢ᩣߣ'pfaA ᩣߩ㜞࿶ਅߦ߅ߌࠆჇᱺ

࿑ 4. S. violacea ߩ㊁↢ᩣߣ'pfaA ᩣߩ㜞࿶ਅߦ߅ߌࠆ⤑‛ᕈ(A) ⤑ߩ⒎ᐨ࿃ሶ S ߩ࿶ജଐሽᕈ (B) ⢽⾰ࠕࠪ࡞㎮ߩ࿁ォ᜛ᢔଥᢙ Dw ߩ࿶ജଐሽᕈ

87

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)6㧚̌⤑ߩᵹേᕈ̍߇ᜬߟᗧ๧
 Homeoviscous adaptation ߪ‫ޔ‬㜞࿶ജ߿ૐ᷷ⅣႺߢ߽⤑ߩ☼ᕈ㧔ᵹേᕈ㧕ࠍ৻ቯߦ଻ߟㆡᔕࠍᗧ๧ߒ‫ޔ‬৻⥸ߦࠕࠪ࡞㎮ߩਇ㘻๺ᐲߢ⺰ߓࠄࠇࠆ‫ޕ‬ᷓᶏߦ᫮߻ᅢ࿶ᕈ⚦⩶ߢ޽ࠆ߇ࠁ߃‫ޔ‬S. violacea DSS12 ߩ⚦⢩⤑ߪߐߙᵹേᕈߦን߻ߦ㆑޿ߥ޿ߣ⠨߃ࠄࠇߡ߈ߚ‫ ߇ࠈߎߣޕ‬HP-TRFAM ߦࠃࠆ⑳ߚߜߩ᷹ቯߪ‫ޔ‬S.
violacea DSS12 ߩ⤑߇㕖Ᏹߦ೰⋥ߢᵹേᕈߦਲߒ޿
ߎߣࠍ᣿ࠄ߆ߦߒߚ‫ޕ‬ታ㓙ߦ᷹ቯߒߡߺߡೋ߼ߡࠊ
߆ߞߚߎߣߢ޽ࠆ‫⁁ߩ⤑ߩߘߦࠄߐޕ‬ᘒߪ 150 MPa
߹ߢട࿶ߒߡ߽ߐ߶ߤᄌൻߒߥ޿‫ޕ‬ᓥߞߡ‫⤑ޔ‬ᵹേᕈߩ⛘ኻ୯߇㊀ⷐߥߩߢߪߥߊ‫ޔ‬࿶ജࠍ߆ߌߡ߽̌⤑߇೰⋥ߩ߹߹ߘࠇએ਄ᄌൻߒߥ޿̍ߎߣߩᣇ߇ᗧ๧
ࠍᜬߟߩߢߥ޿߆ߣ⑳ߚߜߪ⠨߃ߡ޿ࠆ‫ߚߒ߁ߎޕ‬ࠦࡦ࠮ࡊ࠻߇ᨐߚߒߡ᥉ㆉ⊛ߥߩ߆‫੹ޔ‬ᓟ‫ޔ‬ᷓᶏ↱᧪ߩ᭽‫ߥޘ‬ᓸ↢‛⚦⢩⤑ࠍ HP-TRFAM ߢ⸃ᨆߔࠆ
ߎߣߢ╵߃߇⷗ߟ߆ࠆߦ㆑޿ߥ޿‫ޔࠄ߇ߥߒ߆ߒޕ‬߹ߛᧄ⾰⊛ߥ໧㗴߇ᱷߐࠇߡ޿ࠆ‫․ࠍ⤑ޔߪࠇߘޕ‬ᓽߠߌࠆ⒎ᐨ࿃ሶ S ߿࿁ォ᜛ᢔଥᢙ Dw‫ࠇߘߦࠄߐޔ‬ࠄߩ࿶ജଐሽᕈ߇‫ޔ‬ታ㓙ߦߤ߁޿ߞߚ↢‛ᯏ⢻ߣ⋥ធ࡝ࡦࠢߒߡ޿ࠆߩ߆‫߫߃଀ޔ‬ᩕ㙃Ḯャㅍ૕ߩᵴᕈ
߿ࠗࠝࡦ࠴ࡖࡀ࡞ߩᯏ⢻ߥߩ߆‫ߩ⾰‛ࠆߔ੺ࠍ⤑ޔ‬න⚐᜛ᢔߥߩ߆‫⚦ߪ޿ࠆ޽ޔ‬⢩ಽⵚⵝ⟎ߩᯏ⢻ߥߩ߆‫⼂ࠍࠄࠇߘޔ‬೎ߔࠆᄙ㕙⊛ߥࠕࡊࡠ࡯࠴߇੹ᓟᔅ
ⷐߢ޽ࠈ߁‫ޕ‬

4)

5)

6)

7)

⻢ㄉ
 ᧄ⎇ⓥߪਥߦ⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ߢⴕࠊࠇߚ߽ߩߢߔ‫౒ޕ‬ห⎇ⓥ⠪ߩ⥓੗ߌ޿ሶ᳁‫ޔ‬㐿ବ᮸ඳ჻‫ޔ‬ട⮮ජ᣿ඳ჻‫ޔ‬⢻ᧁ⵨৻ඳ჻‫ߦ߮ࠄߥޔ‬੩ㇺᄢቇൻቇ⎇ⓥᚲߩᎹᧄ⚐ඳ჻ߣᩙේ㆐ᄦඳ჻‫ⵝޔ‬⟎㐿⊒ߦ߅޿ߡߪࠪࡦ࡮ࠦ࡯ࡐ࡟࡯࡚ࠪࡦ␠ߩ᧻ᧄ
㓷శඳ჻ߣ᳗Ỉ⋿᳁‫‛⤑ޔߡߒߘޔ‬ᕈߦ㑐ߔࠆ᦭⋉ߥߏഥ⸒ࠍ㗂޿ߚᓼፉᄢቇߩ᧻ᧁဋඳ჻ߣ㊄ຠ᣽ᔒඳ჻ߦᗵ⻢↳ߒ਄ߍ߹ߔ‫ߪ࡯ࡘࡆ࡟࠾ࡒᧄޔ߅ߥޕ‬‫ޟ‬㜞࿶ജߩ⑼ቇߣᛛⴚ‫⃻ߢ⹹ޠ‬࿷ශ೚ਛߩ╩⠪ߩ✚⺑‫ޟ‬㜞࿶ജਅߦ߅ߌࠆᓸ↢‛⚦⢩⤑ߩേ⊛᭴ㅧ‫ࠍޠ‬రߦ✬㓸ߒၫ╩ߒߚ߽ߩߢߔ‫ޕ‬

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Journal of Japanese Society for Extremophiles (2012) Vol.11, 90-95

Honda K.a,b

Construction of synthetic metabolic pathways
using thermo-tolerant biocatalytic modules⠴ᾲᕈ㉂⚛ࡕࠫࡘ࡯࡞ࠍ↪޿ߚวᚑઍ⻢Ꮏቇ߳ߩ᜸ᚢ
ᄢ㒋ᄢቇᄢቇ㒮Ꮏቇ⎇ⓥ⑼‫ޔ‬JST ߐ߈߇ߌᧄ↰ ቁ␭aDepartment of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka
565-0871, Japanb
JST PRESTO
Corresponding author: Kohsuke Honda, honda@bio.eng.osaka-u.ac.jp
Phone: +81-6-6879-7438, Fax: +81-6-6879-7439
Received: September 13, 2012 / Accepted: November 6, 2012㧝㧚ߪߓ߼ߦ ࡃࠗࠝΆᢱ↢↥ࠍ฽߻ฦ⒳⊒㉂↢↥ࡊࡠ࠮ࠬߩ㜞ല₸ൻࠍ⋡ᜰߔ߁߃ߢ‫ޔ‬ઍ⻢Ꮏቇࠍ೑↪ߒߚᓸ↢‛ࠥࡁࡓߩᡷᄌߪ‫߿੹ޔ‬ᰳ߆ߔߎߣߩߢ߈ߥ޿⎇ⓥ࡮㐿⊒ᚢ⇛ߩ߭ߣߟߣߥࠅߟߟ޽ࠆ 8)‫৻ߩߘޕ‬ᣇ‫ޔ‬ઍ⻢Ꮏቇ⊛ᚻᴺߦၮߠ߈⸳⸘࡮ዉ౉ߒߚᡷᄌ߇‫ᦼޔ‬ᓙߤ߅ࠅߩ↢↥ᕈะ਄ߦ⚿߮ઃ߆ߥ޿ߣ޿߁੐଀߽ዋ
ߥߊߥ޿‫ޕ‬ᡷ߼ߡ⠨߃ߡߺࠇ߫ߦߴ߽ߥ޿ߎߣߛ߇‫ޔ‬ᓸ↢‛ߦߣߞߡᦨ߽㊀ⷐߥ࠲ࠬࠢߣߪ‫࡞࡯ࡁ࠲ࠛޔ‬↢↥ߢ߽ࠕࡒࡁ㉄⊒㉂ߢ߽ߥߊ‫⥄ޔ‬り߇⟎߆ࠇߚⅣႺਅߢ޿߆ߦ↢ሽ࡮Ⴧᱺࠍ⛽ᜬߔࠆ߆ߦ޽ࠆ‫ߚߒޕ‬߇ߞߡ‫․߇ࠇࠊࠇࠊޔ‬ቯߩઍ⻢↥‛ߩ↢↥ᕈჇᄢࠍ⋡⺰ࠎߢᓸ↢‛ࠥࡁࡓߦੱὑ⊛ᡷᄌࠍട߃ࠃ߁ߣ߽‫ޔ‬ߘߩᡷᄌ߇ᓐࠄߩ↢ሽ࡮Ⴧᱺߦߣߞߡਇ೑ߦ௛ߊ߽
ߩߢ޽ߞߚ႐ว‫ޔ‬૗ࠄ߆ߩ࠮࡯ࡈ࠹ࠖ࡯ࡀ࠶࠻ࡢ࡯
ࠢ߇⊒േߒ‫ޔ‬ᡷᄌߩലᨐ߇ᛂߜᶖߐࠇࠆߎߣ߇޽ࠆ‫ޕ‬⊒ዷ⪺ߒ޿ฦ⒳ߩ-omics ᛛⴚࠍ㚟૶ߒ‫↢ޟࠆࠁࠊ޿ޔ‬๮ߩࡠࡃࠬ࠻ᕈ‫ߩޠ‬ේ࿃ߣߥߞߡ޿ࠆ৻ㅪߩࡀ࠶࠻
ࡢ࡯ࠢࠍࠃࠅᷓߊℂ⸃ߒ೙ᓮߒࠃ߁ߣ޿߁⹜ߺ߇‫ޔ‬ㄭᐕߩઍ⻢Ꮏቇಽ㊁ߦ߅ߌࠆầᵹߩ߭ߣߟߣߥߞߡ޿ࠆ 5)‫ޕ‬ ৻ᣇ‫⪺ޔ‬⠪ࠄߩࠣ࡞࡯ࡊߢߪ‫⚦ߚ߈↢ޔ‬⢩ߩઍ⻢⚻〝ࠍᡷᄌߔࠆߩߢߪߥߊ‫࡞࡯ࡘࠫࡕޔ‬ൻߒߚઍ⻢㉂⚛ࠍ in vitro ߢછᗧߦ⚵ߺวࠊߖ‫․ߦ↥↢⾰‛ޔ‬ൻߒߚ⚻〝ࠍౣ᭴▽ߔࠆߎߣߢ‫ޔ‬ᣢሽߩઍ⻢Ꮏቇ߇ᛴ߃ࠆ໧㗴ࠍస᦯ߔࠆߎߣࠍ⹜ߺߡ޿ࠆ‫ޕ‬

Abstract Metabolic engineering has emerged as apractical alternative to conventional chemical conversion
particularly for biocommodity production processes.
However, this approach is often hampered by as yet
unidentified inherent mechanisms of natural metabolism.
To overcome this limitation, we developed a novel,simple technology, designated as synthetic metabolicengineering, for freely designing a chimeric metabolic
pathway.
In this technology, genes encoding
thermophilic enzymes were overexpressed in amesophilic host (e.g., Escherichia coli), and therecombinant cells were heated to inactivate mesophilic
indigenous enzymes.
These operations enable a
one-step preparation of highly selective and stablebiocatalytic modules, and an in vitro synthetic metabolic
pathway can be readily constructed through the rational
combination of these modules.
Owing to their
independence from biological activities of living cells,
synthetic pathways no longer play physiological roles innatural metabolism involving the energy generation andenergy-consuming synthesis of biomolecules; thus, thebespoke pathway specialized for chemical conversion
can be designed. In this mini-review, several examplesof the chemical production systems developed usingsynthetic metabolic engineering are shown and thefeasibility of this new technology is discussed.
Keywords: synthetic metabolic engineering, pathway
design, thermo-tolerant enzyme, thermophile, archaea,㧞㧚วᚑઍ⻢Ꮏቇ ᓟㅀߔࠆࠃ߁ߦ‫ޔ‬න㔌㉂⚛ࠍ⚵ߺวࠊߖߚ in vitroߢߩઍ⻢⚻〝ߩ᭴▽ߣߎࠇࠍ↪޿ߚ‛⾰↢↥ߩ଀ߪ90

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ߡઍ⻢⚻〝ࠍ᭴▽ߔࠆ৻ㅪߩ⠴ᾲᕈ㉂⚛ࠍࡕࠫࡘ࡯
࡞ൻߒ‫ࠍࠄࠇߎޔ‬છᗧߦ⚵ߺวࠊߖࠆߎߣߦࠃߞߡ
ൻቇຠ↢↥ߩߚ߼ߩੱᎿઍ⻢⚻〝ࠍࠝࡦ࠺ࡑࡦ࠼ߢ
᭴▽ߔࠆߎߣ߇น⢻ߥࠆ㧔࿑㧝㧕‫ޔ߫ࠇߥ߁⸒ߡ߃଀ޕ‬ᓥ᧪ߩઍ⻢Ꮏቇ߇‫ޔ‬ᣢሽߩ࿾࿑㧔ઍ⻢⚻〝㧕ߩ਄ߦᣂߚߥ㆏ࠍᦠ߈⿷ߒߚࠅ‫ߪ޿ࠆ޽ޔ‬ਇⷐߥᮮ㆏ࠍᶖ
ߒ෰ࠆߎߣߢ಴⊒࿾㧔ၮ⾰㧕߆ࠄ⋡⊛࿾㧔↢↥‛㧕߳ߩ⚻〝ࠍᦨ⍴ൻߔࠆᚻᴺߛߣߔࠆߥࠄ߫‫ᧄޔ‬ᴺߪ
⊕⚕ߩࠠࡖࡦࡃࠬ਄ߦࡈ࡝࡯ࡂࡦ࠼ߢᣂߚߥ࿾࿑ࠍ
࠺ࠩࠗࡦߒࠃ߁ߣ޿߁߽ߩߢ޽ࠆ‫⪺ޕ‬⠪ࠄߩࠣ࡞࡯
ࡊߢߪ‫ᧄޔ‬ᴺࠍ‫ޟ‬วᚑઍ⻢Ꮏቇ‫ߣޠ‬ฬઃߌ‫ޔ‬᭽‫ߥޘ‬ൻቇຠ↢↥߳ߩㆡ↪ࠍ⹜ߺߡ޿ࠆ‫ᧄޕ‬Ⓜߢߪ‫ࠇࠊޔ‬ࠊࠇ߇ߎࠇ߹ߢߦขࠅ⚵ࠎߛวᚑઍ⻢Ꮏቇߦࠃࠆ߽
ߩߠߊࠅߩ੐଀ࠍ⚫੺ߒ‫ߩߘޔ‬น⢻ᕈߣ੹ᓟขࠅ⚵߻ߴ߈⺖㗴ߦߟ޿ߡ⼏⺰ࠍߒߚ޿‫ޕ‬ߎࠇ߹ߢߦ߽ᄙߊߩႎ๔߇ߥߐࠇߡ޿ࠆ‫ޔߒ߆ߒޕ‬ߎߎߢ໧㗴ߣߥࠆߩߪⶄᢙߩઍ⻢㉂⚛ࠍ೽෻ᔕࠍ઻
ࠊߥ޿࡟ࡌ࡞ߦ߹ߢ♖⵾ߔࠆ૞ᬺߩᾘ㔀ߐߦ޽ࠆ‫ޕ‬╩⠪ࠄߪߎߩᾘ㔀ߐࠍ࿁ㆱߔࠆߚ߼‫ޔ‬㧔⿥㧕ᅢᾲ⩶ߦ
↱᧪ߔࠆ⠴ᾲᕈ㉂⚛ㆮવሶߦ⌕⋡ߒߚ‫ޔߕ߹ޕ‬⠴ᾲᕈ㉂⚛ㆮવሶࠍᄢ⣺⩶ߥߤߩਛ᷷ᕈኋਥᓸ↢‛ౝߢㆊ೾⊒⃻ߐߖࠆ‫ޕ‬ᓧࠄࠇߚ⚵឵߃⩶ࠍ 70oC ⒟ᐲߩᾲಣℂߦଏߒߚᓟ‫⸅ޔ‬ᇦߣߒߡߘߩ߹߹೑↪ߔࠆ‫ߎޕ‬ߩ⚿ᨐ‫ޔ‬ኋਥ↱᧪㉂⚛ߩᄢㇱಽ߇ᾲᄌᕈߦࠃࠅਇน
ㅒ⊛ߦᄬᵴߒ‫⵾♖ޔ‬㉂⚛ߣห࡟ࡌ࡞ߩ㜞޿ㆬᛯᕈࠍ
᦭ߒߚ↢૕⸅ᇦ߇ኈᤃߦᓧࠄࠇࠆ‫ߚ߹ޕ‬ᄙߊߩਛ᷷
ᕈᓸ↢‛ߩ⚦⢩ᄖ⤑᭴ㅧߪᾲಣℂߦࠃࠅ⣀ᒙൻߐࠇࠆߚ߼‫ޔ‬ᓸ↢‛ᄌ឵෻ᔕߢߒ߫ߒ߫໧㗴ߣߥࠆၮ⾰࡮↢ᚑ‛ߩ⤑ㅘㆊᓞㅦ߇⸃ᶖߐࠇࠆ‫࡯ࡠࡊࠕᧄޕ‬࠴ߪ⇣⒳ኋਥౝߢߩᯏ⢻⊛⊒⃻ߐ߃น⢻ߢ޽ࠇ߫‫ޔ‬޽ࠄࠁࠆ⠴ᾲᕈ㉂⚛ߦኻߒߡㆡ↪น⢻ߢ޽ࠆ‫ޕ‬ᓥߞ࿑ 1 วᚑઍ⻢Ꮏቇߩᛛⴚ᭎ⷐ㧚91

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)‫ ࠄ߆㉄ࡦ࡝࡝ࡐޕ‬ADP ߳ߩ࡝ࡦ㉄ၮォ⒖෻ᔕࠍ⸅ᇦߔࠆ⠴ᾲᕈࡐ࡝࡝ࡦ㉄ࠠ࠽࡯࠯㧔PPK㧕ࠍ↪޿ߚ
ATP ౣ↢♽ 3)ߩሽ࿷ਅߢ‫ޔ‬DERA ࠍ฽߻ 5 ⒳ߩ㉂⚛
⟲ࠍࠕ࠶࠮ࡦࡉ࡞ߒ‫ ߩࠄ߆ࠬ࡯࠻ࠢ࡞ࡈޔ‬DR5P ↢↥⚻〝ࠍ᭴▽ߒߚ㧔࿑ 2㧕‫ޕ‬ᒰೋ‫ޔ‬ฦ㉂⚛ߩᦨㆡ෻ᔕ
᷷ᐲߦೣߒ‫ޔ‬70oC ߢߩ෻ᔕࠍታᣉߒߚߣߎࠈ‫ޔ‬෻ᔕ㐿ᆎ߆ࠄ 30 ಽ㑆ߢ 10 mM ߩࡈ࡞ࠢ࠻࡯ࠬ߆ࠄ 1.2
mM ⒟ᐲߩ DR5P ߩ↢↥߇⏕⹺ߐࠇߚ‫ߎޔߒ߆ߒޕ‬ࠇࠄߪᤨ㑆ߩ⚻ㆊߣߣ߽ߦᷫዋߒ‫ޔ‬෻ᔕ㐿ᆎ߆ࠄ 2ᤨ㑆߶ߤߢᬌ಴㒢⇇ࠍਅ࿁ߞߚ‫ߪࠇߎޕ‬ઍ⻢⚻〝ߩਛ㑆૕ߢ޽ࠆ GAP ߅ࠃ߮ߘߩ⇣ᕈ૕ߢ޽ࠆࠫࡅ࠼
ࡠࠠࠪࠕ࠮࠻ࡦ࡝ࡦ㉄㧔DHAP㧕߇ᾲߦኻߒߡਇ቟
ቯߢ޽ࠆߎߣߦ⿠࿃ߔࠆ‫ޕ‬DERA ߦࠃࠅ⸅ᇦߐࠇࠆ
෻ᔕߪนㅒ⊛ߦㅴⴕߔࠆߚ߼‫ޔ‬GAP ߩᾲಽ⸃ߦ઻޿‫ޔ‬෻ᔕᐔⴧ߇ࠕ࡞࠼࡯࡞ಽ⸃෻ᔕ஥ߦࠪࡈ࠻ߒ‫ޔ‬ઍ⻢ࡈࡠ࡯ߩㅒᵹ߇↢ߓߡߒ߹߁‫↥↢߼ߚߩߎޕ‬ታ㛎ߪ
PPK ߣ 2 ⒳㘃ߩࠠ࠽࡯࠯ߦࠃࠆࡈ࡞ࠢ࠻࡯ࠬ 1,6-ࡆ
ࠬ࡝ࡦ㉄㧔FBP㧕ߩ↢↥߹ߢࠍ 70oC ߢⴕߞߚᓟ‫ޔ‬ᱷࠅߩ㉂⚛ࡕࠫࡘ࡯࡞ࠍᷝടߒ‫ޔ‬30oC ߦߡᒁ߈⛯߈෻ᔕࠍታᣉߔࠆߣ޿߁ 2 Ბ㓏ߦಽߌߡታᣉߒߚ‫ߩߎޕ‬⚿ᨐ‫ޔ‬10 mM ߩࡈ࡞ࠢ࠻࡯ࠬ߆ࠄ 5.5 mM ߩ DR5P
ࠍ↢↥ߔࠆߎߣ߇น⢻ߢ޽ߞߚ‫ޕ‬㧟㧚วᚑઍ⻢Ꮏቇߦࠃࠆᩭ㉄วᚑਛ㑆૕ߩ↢↥ DNA ߩ↢วᚑߪࠣ࡞ࠦ࡯ࠬࠍ಴⊒‛⾰ߣߒߚ႐ว‫ࠬ࡯ࡏ࡝ޔ‬㛽ᩰߩวᚑߣߘߩㆶరࠍ฽߻ 20 ⒳㘃એ਄ߩ㉂⚛෻ᔕ߇㑐ਈߔࠆⶄ㔀ߥวᚑ⚻〝ࠍ⚻ࠆ‫৻ޕ‬ᣇ‫ޔ‬ㅢᏱߩ↢⚦⢩ౝߢߪࡑࠗ࠽࡯⚻〝ߣߒߡߒ߆ᯏ
⢻ߒߡ޿ߥ޿߽ߩߩ‫ߒߣ〝⚻ࠆߥ⇣ߊోߪߣࠇߎޔ‬ߡ࠺ࠝࠠࠪ࡝ࡏ࡯ࠬࠕ࡞࠼࡜࡯࠯㧔DERA㧕߇⸅ᇦߔࠆࠣ࡝࠮ࡠࠕ࡞࠺ࡅ࠼-3-࡝ࡦ㉄㧔GAP㧕ߣࠕ࠮࠻ࠕ࡞࠺ࡅ࠼ߩࠕ࡞࠼࡯࡞❗วߦࠃࠆ࠺ࠝࠠࠪ㛽ᩰߩ⋥ធᒻᚑࠍ⚻ࠆวᚑ⚻〝߇⍮ࠄࠇߡ޿ࠆ‫ߪ〝⚻ᧄޕ‬࡝ࡏ࡯ࠬࠍ⚻ࠆ‫ߦޠ〝⚻࡯ࡖࠫࡔޟ‬Ყߴߡዋᢙߩ㉂⚛⟲㧔11 ⒳㘃㧕ߦࠃߞߡ᭴ᚑߐࠇߡ޿ࠆ‫ޕ‬ᚲᦸߩઍ⻢⚻〝ࠍ⥄࿷ߦౣ᭴▽ߢ߈ࠆวᚑઍ⻢Ꮏቇ⊛ࠕࡊࡠ࡯࠴ࠍᜬߞߡߔࠇ߫‫ࠍ〝⚻࡯࠽ࠗࡑߥ߁ࠃߩߎޔ‬Ⓧᭂ⊛ߦᵴ↪ߒߚ‛⾰↢↥߇น⢻ߣߥࠆ‫ޕ‬ ⎇ⓥ㐿ᆎᒰᤨ‫ߦߢߔޔ‬ਛ᷷ᕈ DERA ࠍㆊ೾⊒⃻ߐߖߚᄢ⣺⩶ߣ GAP ଏ⛎Ḯߣߒߡ㉂Უߩ⸃♧♽ࠍࠞ࠶ࡊ࡝ࡦࠣߔࠆߎߣߦࠃࠅ‫ޔ‬ᩭ㉄วᚑਛ㑆૕ߢ޽ࠆ2’-࠺ࠝࠠࠪ࡝ࡏ࡯ࠬ 5-࡝ࡦ㉄㧔DR5P㧕ࠍ↢↥ߐߖ
ࠆᚻᴺ߇ႎ๔ߐࠇߡ޿ߚ߇ 2)‫ߥߦࠇߎ߽ࠇࠊࠇࠊޔ‬ࠄ޿ห⚻〝ࠍ᭴▽ߔࠆᔅⷐᦨዊ㒢ߩ⠴ᾲᕈ㉂⚛߆ࠄߥࠆวᚑઍ⻢⚻〝ࠍ᭴▽ߒ DR5P ↢↥ታ㛎ࠍⴕߞߚ1)

࿑ 2 ࡈ࡞ࠢ࠻࡯ࠬ߆ࠄߩ DR5P ↢↥ߩߚ߼ߩวᚑ⚻〝࠺ࠩࠗࡦ 1)㧚
ߩ෻ᔕߢ‫ޔ‬ℂ⺰෼₸ߢ޽ࠆ 2 M㧔9%㧕ߦㄼࠆࠛ࠲ࡁ
࡯࡞ࠍ↢↥ߔࠆߎߣߦᚑഞߒߡ߅ࠅ‫ޔ‬෻ᔕㅦᐲߣኻ
ၮ⾰෼₸ߩߺߢᲧセߔࠇ߫‫ޔ‬ㅢᏱߩ⊒㉂ᴺࠍಒ㚧ߔࠆ↢↥ᕈ߇ᓧࠄࠇߡ޿ࠆ‫ߚ߹ޕ‬ㄭᐕߢߪ‫ޔ‬ጤ↰ߣᅏߦࠃࠅ⠴ᾲᕈ㉂⚛ࠍ↪޿ߚࠛ࠲ࡁ࡯࡞↢↥߇ႎ๔ߐࠇߡ޿ࠆ 4)‫ޕ‬ᅢᾲ⩶ࠍㆮવሶ࠰࡯ࠬߣߔࠆߎߣߢ‫ޔ‬㉂⚛ߩ቟ቯᕈ߇ᩰᲑߦჇߒߚߎߣߦട߃‫࠹ࠬ⵾♖ޔ‬࠶ࡊࠍᄢ᏷ߦ◲⇛ൻߐߖࠆߎߣ߇น⢻ߣߥߞߡ߅ࠅ‫ޔ‬㧠㧚วᚑઍ⻢Ꮏቇߦࠃࠆࠠࡔ࡜ဳ⸃♧♽ߩ᭴▽ ਄ߢ߽ㅀߴߚ߇‫ޔ‬න㔌㉂⚛ࠍ↪޿ߚ in vitro ߢߩઍ⻢⚻〝ߩౣ᭴▽ߣߎࠇࠍ↪޿ߚ‛⾰↢↥ߩ⹜ߺߪ‫ޔ‬ߎࠇ߹ߢߦ߽ᢙᄙߊߩႎ๔߇ߥߐࠇߡ޿ࠆ‫ޕ‬ฎߊߪ
1985 ᐕߦ Welch ߣ Scopes ߦࠃߞߡႎߓࠄࠇߚ㉂Უ
↱᧪ߩ⸃♧♽㉂⚛⟲߅ࠃ߮ ATPase ࠍ↪޿ߚࠣ࡞ࠦ
࡯ࠬ߆ࠄߩࠛ࠲ࡁ࡯࡞↢↥ߩ଀߇᜼ߍࠄࠇࠆ 9)‫ᧄޕ‬ႎ๔ߦࠃࠇ߫‫ޔ‬1 M㧔18%㧕ߩࠣ࡞ࠦ࡯ࠬ߆ࠄ 8 ᤨ㑆
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)߫ࠣ࡞ࠦ࡯ࠬ 1 ಽሶ޽ߚࠅ 2 ಽሶߩ ATP ߇↢↥ߐࠇࠆ㧔ߔߥࠊߜ ADP 2 ಽሶ߇ᶖ⾌ߐࠇࠆ㧕‫৻ޕ‬ᣇ‫৻ޔ‬ㇱߩࠕ࡯ࠠࠕߦߪ ADP ଐሽဳࠠ࠽࡯࠯ߥߤ࡙࠾࡯
ࠢߥ⵬㉂⚛ⷐ᳞ᕈࠍ᦭ߒߚ⌀ᩭ࡮ේᩭ↢‛ߦߪ⷗ࠄ
ࠇߥ޿㉂⚛߆ࠄߥࠆᄌᒻ EM ⚻〝ߩሽ࿷߇⍮ࠄࠇߡ޿ࠆ 7)‫ޕ‬ේᩭ࡮⌀ᩭ↢‛ဳ EM ⚻〝ߦ߅޿ߡ GAP ߆ࠄ 3-ࡎࠬࡎࠣ࡝࠮࡝ࡦ㉄㧔3-PG㧕߳ߩᄌ឵ߪ‫ޔ‬GAP࠺ࡅ࠼ࡠࠥ࠽࡯࠯㧔GAPDH㧕ߦࠃࠆ࡝ࡦ㉄ଐሽဳߩ⣕᳓⚛෻ᔕߣࡎࠬࡎࠣ࡝࠮࡝ࡦ㉄ࠠ࠽࡯࠯㧔PGK㧕ߦࠃࠆ⣕࡝ࡦ㉄ൻ෻ᔕߩ 2 Ბ㓏߆ࠄᚑࠅ┙ߞߡ߅ࠅ‫ޔ‬ߎߩ৻ㅪߩ෻ᔕߢ GAP 1 ಽሶ޽ߚࠅ 1 ಽሶߩ ATP߇↥↢ߐࠇࠆ‫ߦࠇߎޕ‬ኻߒ‫৻ޔ‬ㇱߩࠕ࡯ࠠࠕߪ‫࡝ޔ‬ࡦ㉄㕖ଐሽ⊛ߦ GAP ࠍ⣕᳓⚛ߔࠆ GAP ࡈࠚ࡟࠼ࠠ
ࠪࡦ㉄ൻㆶర㉂⚛㧔GAPOR㧕߿ non-phosphorylating
GAPDH㧔GAPN㧕ߦࠃࠅ GAP ߆ࠄ 1 Ბ㓏ߩ෻ᔕߢ⋥ធ 3-PG ߳ߩᄌ឵ࠍⴕ߁‫ޕ‬GAPN ߇⸅ᇦߔࠆ෻ᔕ
ߢߪ ATP ↢↥ߪ઻ࠊߥ޿ߚ߼‫ޔ‬ේᩭ࡮⌀ᩭ↢‛ဳ
EM ⚻〝ਛߩ GAPDH ߅ࠃ߮ PGK ࠍࠕ࡯ࠠࠕ↱᧪ߩGAPOR ߿ GAPN ߦ⟎឵ߔࠆߎߣߢ ATP ෼ᡰߩว⥌ߒߚࠠࡔ࡜ဳ EM ⚻〝ࠍ᭴▽ߔࠆߎߣ߇น⢻ߣߥࠆ
㧔࿑ 3㧕
‫ޕ‬

ߎߩࠕࠗ࠺ࠕߪᚒ‫ߩޘ‬ᛛⴚࠍ┙᩺ߔࠆ਄ߢ߽ᰳ߆ߖ
ߥ޿߽ߩߢ޽ߞߚ‫ߪ♽ᧄޕ‬૛೾ ATP ߩౣ↢෻ᔕࠍ฽߹ߥ޿ߚ߼‫ޔ‬㜞Ớᐲၮ⾰ߩ೑↪ߦߪㆡߐߥ޿߇‫ޔ‬5mM ߩࠣ࡞ࠦ࡯ࠬ߆ࠄ 9 mM ⒟ᐲߩࠛ࠲ࡁ࡯࡞↢↥߇น⢻ߢ޽ࠅ‫ޔ‬ኻၮ⾰෼₸ߩ㜞ߐߪ Welch ߣ Scopesߩࠪࠬ࠹ࡓߣห╬ߢ޽ࠆ‫ޕ‬ߚߛߒ‫ߩࠄࠇߎޔ‬ႎ๔଀ߢౣ᭴▽ߐࠇߚઍ⻢⚻〝ߪ‫ޔ‬޿ߕࠇ߽ᄤὼߩࠛ࠲ࡁ࡯࡞⊒㉂⚻〝ࠍߘߩ߹߹ᮨ୮ߒ ߚ ߽ ߩ ߢ ޽ ࠆ ‫ ޕ‬ઍ ⻢ ߩ ↢ ℂ ⊛ ᗧ ⟵ ߇ ATP ߿
NAD(P)H ߥߤߩ⵬㉂⚛ࠍㅢ⽻ߣߒߚࠛࡀ࡞ࠡ࡯߿
ㆶరജߩ₪ᓧ㧔⇣ൻઍ⻢㧕ߥࠄ߮ߦ‫ߢߎߎޔ‬ᓧࠄࠇߚࠛࡀ࡞ࠡ࡯ࠍ೑↪ߒߚ⚦⢩᭴ᚑ‛⾰ߩวᚑ㧔หൻઍ⻢㧕ߦ޽ࠆὐࠍ⠨ᘦߔࠇ߫‫ޔ‬ᄤὼߩઍ⻢⚻〝ߩ৻ㇱࠍή૞ὑߦ‫࡯ࡇࠦޟ‬㧒ࡍ࡯ࠬ࠻‫ߩߌߛߚߒޠ‬วᚑ⚻〝ߢߪ‫⵬ޔ‬㉂⚛ߩㆊਇ⿷ߦࠃࠆ෻ᔕ஗ᱛ߇ㆱߌࠄ
ࠇߥ޿ߎߣߪ⥄᣿ߢ޽ࠆ‫ޕ‬PPK ߿ ATPase ߥߤߩㆡᒰߥ⵬㉂⚛ౣ↢↪㉂⚛෻ᔕߣߩࠞ࠶ࡊ࡝ࡦ߽ࠣ৻ߟ
ߩ⸃᳿╷ߣ޿߃ࠆ߇‫ޔ‬ᓸ↢‛ߩઍ⻢㉂⚛ߩᄙ᭽ᕈࠍ
ᵴ↪ߔࠇ߫‫ߩࠄࠇߎޔ‬ᣇᴺߦ㗬ࠄߥߊߣ߽⵬㉂⚛෼ᡰߩ໧㗴ࠍస᦯ߔࠆߎߣ߇น⢻ߣߥࠆ‫࡞ࠣ߫߃଀ޕ‬ࠦ࡯ࠬ߆ࠄߩࠛ࠲ࡁ࡯࡞↢↥⚻〝ߩ႐ว‫⌀ޔ‬ᩭ࡮ේᩭ↢‛ߦ߅ߌࠆ Embden-Meyerhof㧔EM㧕⚻〝ࠍ⚻ࠇ࿑ 3 ࠠࡔ࡜ဳ⸃♧♽ߩ࠺ࠩࠗࡦ㧚⌀ᩭ/ේᩭ↢‛ဳ EM ⚻〝㧔Ꮐ㧕‫ဳࠕࠠ࡯ࠕޔ‬ᄌᒻ EM ⚻〝㧔ਛ㧕‫߮ࠃ߅ޔ‬╩⠪ࠄ߇᭴▽ߒߚࠠࡔ࡜ဳ EM ⚻〝㧔ฝ㧕‫ޕ‬ᄌᒻ EM ⚻〝ߦ․ᓽ⊛ߥ㉂⚛෻ᔕࠍ⿒⍫ශߢ␜ߒߚ㧚93

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)NAD+ߦࠃࠆ㒖ኂ૞↪ࠍⵍࠆߎߣߥߊ LDH ߣห᭽ߩ෻ᔕࠍ⸅ᇦน⢻ߥ೎ߩ㉂⚛ࠍត⚝ߒ‫㉄ࠧࡦ࡝ޔ‬/੃㉄࠺ࡅ࠼ࡠࠥ࠽࡯࠯㧔MLDH㧕ࠍㆬᛮߒߚ‫⚦ߚ߈↢ޕ‬⢩ࠍ↪޿ߥ޿วᚑઍ⻢Ꮏቇߢߪ‫ޔ‬ォ౮࡮⠡⸶࡟ࡌ࡞ߢߩ⺞▵૞↪ߦᏀฝߐࠇߥ޿‛⾰↢↥߇น⢻ߢ޽ࠆߎߣߪ⸒߁߹ߢ߽ߥ޿߇‫ࠆߥ⇣ߦ߁ࠃߩߎޔ‬ઍ⻢⚻〝㧔޽ࠆ޿ߪ⇣ߥࠆᓸ↢‛㧕ߦ↱᧪ߔࠆ㉂⚛ࠍࠕ࠶࠮ࡦࡉ࡞ߔࠆߎߣߦࠃࠅ࠲ࡦࡄࠢ⾰࡟ࡌ࡞ߢ૞↪ߔࠆࠕࡠࠬ࠹࡝࠶ࠢ⺞▵ࠍ߽࿁ㆱߢ߈ࠆὐߦߪ⌕⋡ߔߴ߈ߢ޽ࠆ‫ޕ‬ ߎ߁ߒߡ᭴▽ߐࠇߚࠠࡔ࡜ဳ EM ⚻〝ࠍ↪޿ߡ‫ޔ‬↢↥ታ㛎ࠍⴕߞߚߣߎࠈ‫ޔ‬෻ᔕ㐿ᆎ߆ࠄ 5 ᤨ㑆⒟ᐲ߹ߢ⛮⛯⊛ߥ੃㉄ߩ⫾Ⓧ߇⹺߼ࠄࠇߚ㧔࿑ 4㧕‫ߩߘޕ‬ᓟ‫↥↢ޔ‬ㅦᐲߥࠄ߮ߦࡕ࡞෼₸ߩૐਅ߇⹺߼ࠄࠇߚ
߇‫ߪࠇߎޔ‬㉂⚛ߩᄬᵴߦࠃࠆ߽ߩߢߪߥߊ‫⵬ޔ‬㉂⚛
ߢ޽ࠆ NAD+߅ࠃ߮ NADH ߩᾲಽ⸃ߦ⿠࿃ߔࠆߎߣ
߇᣿ࠄ߆ߣߥߞߚ‫ޕ‬෻ᔕ㐿ᆎ 5 ᤨ㑆ᓟߦ 1 mM ߩ
NADH ࠍౣᷝടߔࠆߎߣߦࠃࠅ‫ޔ‬෻ᔕㅦᐲߪ࿁ᓳߒ‫ޔ‬
10 ᤨ㑆ߩ෻ᔕߢ 6 mM ߩࠣ࡞ࠦ࡯ࠬ߆ࠄࡕ࡞෼₸100%ߩᄌ឵‫ ߜࠊߥߔޔ‬12 mM ߩ੃㉄ࠍᓧࠆߎߣ߇
น⢻ߣߥߞߚ‫ߩߎޕ‬㓙‫〝⚻ޔ‬ౝߢߩ ATP ࠲࡯ࡦࠝ࡯
ࡃ࡯ᢙߪ 31 ߣ▚಴ߐࠇߚ‫ޕ‬ ߎߎߢ㊀ⷐߣߥࠆߩߪ‫〝⚻ဳ࡜ࡔࠠߥ߁ࠃߩߎޔ‬ߪวᚑઍ⻢Ꮏቇ⊛ᚻᴺࠍ↪޿ߥߌࠇ߫᭴▽ߢ߈ߥ޿ὐߦ޽ࠆ‫ޔߊߥ߽ߢ߹߁⸒ޕ‬EM ⚻〝ߪ‫ߊࠃ߽ᦨޔ‬⎇ⓥߐࠇߚ‫߽ᦨߡߞߣߦ‛↢ߡߒߘޔ‬㊀ⷐߥઍ⻢⚻〝ߩ߭ߣߟߢ޽ࠅ‫ޔ‬ၮ⾰㧔ࠣ࡞ࠦ࡯ࠬ㧕߇᦭ߔࠆ⥄↱↢ᚑࠛࡀ࡞ࠡ࡯ࠍ ATP ߩᒻߢขࠅ಴ߔߎߣࠍ↢ℂ⊛ᓎഀߣߒߡ޿ࠆ‫ޔߜࠊߥߔޕ‬ATP ෼ᡰߩว⥌ߒߚ
㧔ATP ࠍ↢↥ߒߥ޿㧕EM ⚻〝ߪ‫↢ߩߘ߿ߪ߽ޔ‬ℂ⊛ᓎഀࠍᄬߞߡ߅ࠅ‫↢ࠍ〝⚻ဳ࡜ࡔࠠߥ߁ࠃߩߎޔ‬߈ߚᓸ↢‛ౝߢౣ᭴▽ߔࠆߣ޿߁ߎߣߪ‫ޔ‬ขࠅ߽⋥ߐߕᒰ⹥ᓸ↢‛ߩ EM ⚻〝ߩᯏ⢻ࠍήലൻߔࠆߎߣ
ߦઁߥࠄߥ޿߆ࠄߢ޽ࠆ‫ޕ‬ ╩⠪ࠄߪ‫ޔ‬Thermococcus kodakarensis ↱᧪ߩ GAPN
6)ߥࠄ߮ߦ Thermus thermophilus ↱᧪ߩේᩭ↢‛ဳEM ⚻〝㉂⚛⟲ࠍ⚵ߺวࠊߖߚࠠࡔ࡜ဳ EM ⚻〝ࠍ᭴▽ߒ‫ߦ↥↢㉄੃ߩࠄ߆ࠬ࡯ࠦ࡞ࠣߚ޿↪ࠍࠇߎޔ‬ขࠅ⚵ࠎߛ 10)‫ࠆߥߣࡊ࠶࠹ࠬ⚳ᦨߩ〝⚻ᧄޔ߅ߥޕ‬ࡇ࡞ࡆࡦ㉄߆ࠄ੃㉄߳ߩ NADH ଐሽ⊛ㆶర෻ᔕߪ
ㅢᏱ‫࠯࡯࠽ࠥࡠ࠼ࡅ࠺㉄੃ޔ‬㧔LDH㧕ߦࠃߞߡ⸅ᇦ
ߐࠇࠆ߇‫ޔ‬Thermus thermophilus ↱᧪⠴ᾲᕈ LDH ߪ‫ޔ‬Ყセ⊛ૐỚᐲ㧔0.5 mM એ਄㧕ߩ NAD+ሽ࿷ਅߢᒝߊ
㒖ኂࠍฃߌ‫ޔ‬ห⚻〝ౝߢߪ߶ߣࠎߤᯏ⢻ߒߥ޿ߎߣ
߇᣿ࠄ߆ߣߥߞߚ‫ߢߎߘޕ‬Ṷ⠪ࠄߪ‫ޔ‬ห⚦⩶ࠃࠅ

࿑ 4 ࠠࡔ࡜ဳ⸃♧♽ࠍ↪޿ߚࠣ࡞ࠦ࡯ࠬ߆ࠄߩ੃㉄↢↥ 10)㧚੃㉄㧔٤㧕߅ࠃ߮ NAD+/NADH ߩ✚Ớᐲ㧔‫غ‬㧕ߩ⚻ᤨᄌൻࠍ␜ߒߚ㧚෻ᔕ㐿ᆎ 5 ᤨ㑆ᓟߦ 1 mM NADH ࠍᷝടߒߚ႐วࠍࠝ࡟ࡦࠫ‫ޔ‬ਇᷝടߩ႐วࠍ✛ߢ␜ߒߚ㧚ߪ‫↥ߛ߹ߛ߹ޔ‬ᬺᔕ↪น⢻ߥ࡟ࡌ࡞ߦߪ⒟㆙޿‫ߘޕ‬
ߎߢᧄⓂߩᦨᓟߦ‫ޔ‬วᚑઍ⻢Ꮏቇߩࡄࡈࠜ࡯ࡑࡦࠬ
ࠍታ↪࡟ࡌ࡞ߦ߹ߢ᛼ߒ਄ߍࠆߚ߼‫੹ޔ‬ᓟࠊࠇࠊࠇ
߇ขࠅ⚵߻ߴ߈⺖㗴ߦߟ޿ߡ⸅ࠇߚ޿‫ޕ‬ ੹࿁⚫੺ߒߚ⎇ⓥ੐଀ߢߪ‫ⶄޔ‬ᢙߩ⠴ᾲᕈ㉂⚛ࡕࠫࡘ࡯࡞ࠍ⚵ߺวࠊߖࠆ㓙‫ޔ‬ฦ㉂⚛ㆮવሶࠍ୘೎ߦ
⊒⃻ߐߖߚ⚵឵߃ᄢ⣺⩶ࠍᷙวߒߡ૶↪ߒߚ‫ߩߎޕ‬㧡㧚߅ࠊࠅߦ એ਄ߩ⎇ⓥᚑᨐࠍㅢߓߡ‫ޔ‬ઍ⻢⚻〝ߩࡈ࡝࡯࠺ࠩ
ࠗࡦ߿೽↥‛ࠍ઻ࠊߥ޿㧔ߔߥࠊߜࡕ࡞෼₸ 100%ߢߩ㧕‛⾰↢↥߇น⢻ߣߥࠆ‫ߚߞ޿ߣޔ‬ၮᐙࠦࡦ࠮
ࡊ࠻߇߅߅߻ߨታ⸽ߐࠇߚ෻㕙‫ޔ‬ฦ⎇ⓥ੐଀ߦ߅ߌ
ࠆᦨ⚳↢↥‛Ớᐲߥߤ߆ࠄ߽߅ࠊ߆ࠅ޿ߚߛߌࠆߣ
߅ࠅ‫ޔ‬ൻቇຠ↢↥ᛛⴚߣߒߡߩวᚑઍ⻢Ꮏቇߩജଔ
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ᣇ╷ࠍขࠆߎߣߦࠃߞߡ‫ޔ‬୘‫ߩޘ‬㉂⚛ᵴᕈ࡟ࡌ࡞ߩ
ࡈࠔࠗࡦ࠴ࡘ࡯࠾ࡦࠣ߇น⢻ߣߥࠆ෻㕙‫ޔ‬᭴▽ߒߚ޿ઍ⻢⚻〝߇ⶄ㔀ߦߥࠆ߶ߤၭ㙃࡮㓸⩶ߦ߆߆ࠆᚻ
㑆߇ᾘ㔀ߣߥࠆ‫ߦࠇߎޕ‬ኻߒ‫ޔ‬ᔅⷐߥ⠴ᾲᕈ㉂⚛ㆮવሶ⟲ࠍ౒⊒⃻ߐߖߚ⚵឵߃ᩣࠍ⸅ᇦߣߒߡ↪޿ࠆߎߣ߇ߢ߈ࠇ߫‫ޔ‬ၭ㙃ߦⷐߔࠆᚻ㑆ࠍシᷫߔࠆߩߺ
ߥࠄߕ‫ޔ‬ᔅⷐߥ㉂⚛ࠍ⚦⢩ౝߣ޿߁ᓸዊߥⓨ㑆ౝߦ஍࿷ߐߖࠆߎߣߦࠃࠅ‫ޔ‬ઍ⻢ਛ㑆૕ߩ᜛ᢔ〒㔌ࠍ❗
߼‫✚ޔ‬᜝ߩ෻ᔕㅦᐲࠍ㜞߼ࠆ‫” ࠆࠁࠊ޿ޔ‬spatialorganization” ߇น⢻ߦߥࠆߣᦼᓙߢ߈ࠆ‫ޕ‬ ߹ߚ‫ߩ↥↢㉄੃ࠆࠃߦ♽♧⸃ဳ࡜ࡔࠠޔ‬ㆊ⒟ߢ᣿
ࠄ߆ߦߥߞߚ⵬㉂⚛ߩᾲಽ⸃ߩ໧㗴ߪ‫ޔ‬‫ޟ‬㧔⿥㧕ᅢᾲ⩶ߪ޿߆ߦߒߡߎࠇࠄߩ‛⾰ߩ⚦⢩ౝỚᐲࠍ଻ߞߡ
޿ࠆߩߛࠈ߁߆㧫‫߁޿ߣޠ‬ᣂߚߥ⇼໧ࠍࠊࠇࠊࠇߦ
ឭ␜ߔࠆ߽ߩߛߣ߽޿߃ࠆ‫ޕ‬ᓸ↢‛ߩ↢⢒᷷ᐲߣߎࠇࠄߩ⚦⢩ౝߦ߅ߌࠆ⵬㉂⚛↢วᚑㅦᐲߦߪ⋧㑐߇޽ࠆߩߛࠈ߁߆㧫޽ࠆ޿ߪ‫ޔ‬ᅢᾲ⩶ߦߪ‫ᧂޔ‬⍮ߩ⵬㉂⚛቟ቯൻ࿃ሶ߇ሽ࿷ߔࠆߩߛࠈ߁߆㧫ߎࠇࠄߩὐ
ߦߟ޿ߡࠃࠅᷓ޿⍮⷗߇ᓧࠄࠇࠇ߫‫ޔ‬วᚑઍ⻢Ꮏቇߩࡈࠖ࡯ࠫࡆ࡝࠹ࠖ࡯ࠍะ਄ߐߖࠆ਄ߢ߽ᄢ߈ߥ৻
ᱠߣߥࠆߪߕߢ޽ࠆ‫ޕ‬ߘߒߡ‫ߩࠕࠠ࡯ࠕޔ‬ᄌᒻ EM ⚻〝ਛߩ㉂⚛⟲߇ࠠࡔ࡜ဳ⸃♧♽᭴▽ߩߚ߼ߩࠠ࡯࠷࡯࡞ߣߥߞߚࠃ߁ߦ‫ޔ‬⃻࿷߽♖ജ⊛ߦㅴ߼ࠄࠇࠆᅢᾲ⩶ߩઍ⻢⚻〝߅ࠃ߮ߘࠇࠍߟ߆ߐߤࠆ⠴ᾲᕈ㉂⚛⟲ߩ․ᕈ⹏ଔߣ࡜ࠗࡉ
࡜࡝࡯ൻ߇‫ޔ‬วᚑઍ⻢Ꮏቇߦࠃࠆ‫ࠅࠃޔ‬ᄙ᭽ߢ⥄࿷ߥઍ⻢⚻〝࠺ࠩࠗࡦߦᰳ߆ߖߥ޿ߎߣߪ⸒߁߹ߢ߽
ߥ߆ࠈ߁‫ޕ‬

3)

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)Journal of Japanese Society for Extremophiles (2012) Vol.11, 96-104

㪤㪠㪥㪠㩷㪩㪜㪭㪠㪜㪮㩷

Takai K.

What is life?
Lesson from boundary between the habitable and the uninhabitable in the Earth↢๮ߣߪ૗߆㧫г↢๮㧔࿤㧕ߩ㒢⇇߆ࠄ⠨߃ࠆг
⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ ᶏᵗ࡮ᭂ㒢ⅣႺ↢‛࿤㗔ၞ߅ࠃ߮ࡊ࡟ࠞࡦࡉ࡝ࠕࡦࠛࠦࠪࠬ࠹ࡓ࡜ࡏ
㜞੗ ⎇Subsurface Geobiology Advanced Research (SUGAR) Project and Precambrian Ecosystem Laboratory (PEL),
Japan Agency for Marine-Earth Science & Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan
Corresponding author: Ken Takai, kent@jamstec.go.jp
Phone: +81-46-867-9677, Fax: +81-46-867-9715
Received: October 29, 2012 / Accepted: November 7, 2012✜⸒ ࠕ࡝ࠬ࠻࠹࡟ࠬߦᆎ߹ࠆߣߐࠇࠆ‫ޔ‬ᚒ‫⥄ޘ‬りࠍ฽߼ߚ࿾⃿ߦ↢߈᳗ࠄ߃ࠆࡕࡁ‫ࠍ—‛↢ࠅ߹ߟޔ‬ℂ⸃
ߒࠃ߁ߣߔࠆ⍮ߩ૕♽ߣߒߡߩ‫‛↢ޟ‬ቇ‫߈⸃ޔ߇ޠ‬᣿߆ߘ߁ߣߔࠆⓥᭂ⊛ߥ೔㆐ὐߩ৻ߟߪ‫ޔ‬‫↢ޟ‬๮ߣߪ
૗߆‫߁޿ߣޠ‬໧޿ߦኻߒߡ‫⑼ޔ‬ቇ⊛ߥ৻⥸ᕈේℂߣ
ߒߡߩ╵߃ࠍឭ␜ߔࠆߎߣߢ޽ࠈ߁‫╵ߩߘߡߒߘޕ‬߃ߦ೔㆐ߒࠃ߁ߣߔࠆߎߣߪ‫‛↢ޔ‬ቇߛߌߦ⇐߹ࠄߕ‫⑼ࠆࠁࠄ޽ޔ‬ቇߦ߅ߌࠆ‫ੱߪߦࠄߐޔ‬㑆ߩ⍮⊛ᅢᄸᔃߩᩮḮ⊛ߥ᰼᳞ߦၮߠ޿ߚ৻ߟߩ⍮ߩ⋡ᮡߣ⸒߃ࠆߩߢߪߥ޿ߛࠈ߁߆‫৻ߒ߆ߒޕ‬ᣇߢ‫ߩߘޔ‬๮㗴ߦኻߔࠆ⑼ቇࠕࡊࡠ࡯࠴߇᛽⽎⊛߆ߟⷰᔨ⺰⊛ߥ᭎ᔨ߿⠨ኤߦ⇐߹ࠆ㒢ࠅߦ߅޿ߡߪ‫ޔ‬⍮⊛ᅢᄸᔃߩべേᗵ߿㝯ജࠍᄬߞߡߒ߹߁น⢻ᕈุ߽ቯߢ߈ߥ޿‫ޕ‬ ߟ߹ࠅ⪺⠪ߪ‫ޔ‬‫↢ޟ‬๮ߣߪ૗߆‫߁޿ߣޠ‬໧޿ߦ╵߃
ࠆ⑼ቇࠍᒝജߦផㅴߔࠆߚ߼ߦߪ‫ⷰޔ‬ᔨ⺰⊛ߥ᭎ᔨߣߣ߽ߦ‫ޔ‬Ყセ⊛ࠊ߆ࠅ߿ߔ޿ᬌ⸽ߔߴ߈઒⺑ߦၮߠ޿ߚታ⃻น⢻߆ߟ㝯ജ⊛ߥ⎇ⓥ࠹࡯ࡑ߿ࡊࡠࠫࠚ
ࠢ࠻߇ᔅⷐߢߪߥ޿߆ߣ⠨߃ࠆ‫ⷰߥ߁ࠃߩߘޕ‬ὐ߆ࠄୄ⍑ߔࠇ߫‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮߇ሽ࿷ߢ߈ߥ޿ࠃ
߁ߦ⷗߃ࠆᭂ㒢⊛ߥⅣႺߦ߅޿ߡ‫↢ޔ‬๮ᵴേࠍሽ⛯ߔࠆ↢‛‫ࠅ߹ߟޔ‬ᭂ㒢ⅣႺ↢‛—ߩ⎇ⓥߪ‫ޔ‬‫↢ޟ‬๮ߣ
ߪ૗߆‫ࠍޠ‬⠨߃ࠆ਄ߢ৻ߟߩౕ૕⊛ߥᚻ߇߆ࠅߦߥ
ࠆ‫↢ޟ‬๮ߩ㒢⇇‫ߦޠ‬ኻߔࠆᧄ⾰⊛ℂ⸃ࠍਈ߃ࠆ੐߇
ߢ߈ࠆ⎇ⓥಽ㊁ߣ⸒߃ࠃ߁‫ޕ‬ ߹ߚᚒ‫߇ޘ‬໑৻⍮ߞߡ޿ࠆ‫ޟ‬࿾⃿↢๮‫ߥߢߌߛޠ‬ߊ‫ߊࠄߘ߅ޔ‬ቝቮߦ߅޿ߡ߽ᐞᄙߦሽ࿷ߔࠆߣ⠨߃ࠄࠇࠆ࿾⃿ᄖ↢๮ߩ޽ࠅᣇߣߩᲧセࠍㅢߓߡ‫ޔ‬‫↢ޟ‬๮ߣߪ૗߆‫ࠍޠ‬ℂ⸃ߒࠃ߁ߣߔࠆ‫ޟ‬ᄥ㓁♽ߦ߅ߌࠆ࿾
⃿ᄖ↢๮តᩏ‫ߦޠ‬ઍ⴫ߐࠇࠆࠃ߁ߥࠕࠬ࠻ࡠࡃࠗࠝ

Abstract I believe that one of the fundamental goals in
research on extremophiles is to have a general principle
explaining the question, ‘what is life?’. Extremophiles
living in certain boundary biospheres, biospheres present
in fringes between habitable and uninhabitableenvironments in this planet, have greater potentials tooriginate, proliferate, evolve, survive and propagate inpotentially habitable environments in extraterrestrial
bodies and moons in the Universe. Thus, if we havechances to find such extraterrestrial extremophiles in the
future, comparative studies between the terrestrial andthe extraterrestrial extremophiles will provide a
revolutionary insight into understanding what life is.
However, we should establish certain definitions andcriteria to perceive existence and function of life in
future extraterrestrial life exploration. Of particular,
concepts for limits of life and biospheres and habitability
in this planet build the basis. Here, I overview recent
progress in extremophiles’ research to extend our viewon limits of life and biosphere in this planet. In addition,a currently proposed energy mass balance concept of
habitability and the subsequent several estimations forthe realistic minimal energy quantum and quantity tosustain the terrestrial lives are introduced. At last, thedifficulties that we now encounter and the future
prospects are discussed.
Keywords: Extremophiles, limits of life, limits ofbiosphere, energy mass balance, Astrobiologyࠠ࡯ࡢ࡯࠼㧦ᭂ㒢ⅣႺ↢‛‫↢ޔ‬๮ߩ㒢⇇‫↢ޔ‬๮࿤ߩ㒢⇇‫ޔ⺰࡯ࠡ࡞ࡀࠛޔ‬ቝቮ↢‛ቇ

96

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ࡠࠫ࡯ߣ޿߁ᣂߒ޿⎇ⓥ㗔ၞߪ‫⑼ޔ‬ቇ⊛ߦ߽␠ળ⊛ߦ߽ᭂ߼ߡ᳞ᔃജߦንࠎߛ⎇ⓥಽ㊁ߣߒߡߩࡐ࠹ࡦ
ࠪࡖ࡞ࠍ⒁߼ߡ޿ࠆ‫ޕ‬ታ㓙ߦߎࠇ߹ߢߩ⑼ቇ⊛ᚑᨐߦၮߠ޿ߚౕ૕⊛ߥ࿾⃿ᄖ↢๮តᩏ⸘↹ߣߒߡ‫ޔ‬਎⇇ฦ࿖ߢ‫ޔ‬Ἣᤊ‫ᧁޔ‬ᤊߩⴡᤊߢ޽ࠆ࡙࡯ࡠࡄ‫ޔ‬࿯ᤊߩⴡᤊࠛࡦ࠮࡜࠳ࠬ࡮࠲ࠗ࠲ࡦ‫↢ߩޔ‬๮តᩏ߇ផㅴ‫ޔ‬ዷ㐿ߐࠇ‫ޔ‬ᄢ߈ߥᵈ⋡ࠍᶎ߮ߡ޿ࠆ‫ߩࠇߕ޿ޕ‬୥⵬
ⅣႺߦ߅޿ߡ߽‫ߩߘޔ‬૞ᬺ઒⺑ߩၮ⋚ߪ‫(ޔ‬1)↢๮⃻⽎ࠍᡰ߃ࠆ⥄↱ࠛࡀ࡞ࠡ࡯߇චಽߦሽ࿷ߒߡ޿ࠆ
㧔޿ߚ㧕ߎߣ‫(ߡߒߘޔ‬2)ⓨ㑆⊛࡮ᤨ㑆⊛ߥࠛࡀ࡞ࠡ
࡯ࡐ࠹ࡦࠪࡖ࡞ࠍᒻᚑߔࠆ‫ޟ‬඙ಾࠅ᭴ㅧ
(compartment)‫߿ޠ‬ᄙ᭽ߥήᯏ‛߿᦭ᯏ‛෸߮ߘߩൻ
ቇ෻ᔕࠍᡰ߃ࠆ‫ޟ‬ṁᇦ(solvent)‫߇ޠ‬චಽߦሽ࿷ߒߡ
޿ࠆ㧔޿ߚ㧕ߎߣ‫߇ޔ‬ᗐቯߐࠇߡ޿ࠆ‫ޟߡߒߘޕ‬ṁᇦ‫ߦޠ‬㑐ߒߡߪ‫ࠍࡦ࠲ࠗ࠲ޔ‬㒰޿ߡߪ‫ޔ‬‫ޟ‬ᶧ૕ߩ᳓‫ޠ‬߇೨ឭ᧦ઙߣߒߡ᜼ߍࠄࠇߡ޿ࠆ‫ޕ‬ ߟ߹ࠅ‫ޔ‬ᄥ㓁♽ߦ߅ߌࠆ࿾⃿ᄖ↢๮តᩏߣߪ‫ޔ‬‫⥄ޟ‬↱ࠛࡀ࡞ࠡ࡯‫ޟߣޠ‬ᶧ૕ߩ᳓‫ߩޠ‬ሽ࿷ࠍ࿾⃿ᄖ↢๮ሽ࿷ߩ೨ឭ᧦ઙߣߒߡᝒ߃ߡ޿ࠆߣ⸒ߞߡࠃ޿‫ߘޕ‬ߒߡߘࠇߪ࿾⃿ߩᭂ㒢ⅣႺߦ↢ᕷߔࠆ↢‛ߩ⎇ⓥߦ
ࠃߞߡⓍߺ਄ߍࠄࠇߡ߈ߚ࿾⃿↢๮ሽ࿷ߩᔅⷐᦨૐ
೨ឭ᧦ઙߢ޽ࠆ‫ࠎࠈߜ߽ޕ‬࿾⃿ᄖ↢๮ሽ࿷᧦ઙߪ‫ޔ‬࿾⃿ᭂ㒢ⅣႺ↢‛ߩሽ࿷น⢻᧦ઙࠃࠅ߽ᄢ߈ߥࡐ࠹
ࡦࠪࡖ࡞ࠍ᦭ߒߡ޿ࠆน⢻ᕈ߇චಽ⠨߃ࠄࠇࠆ‫ߒޕ‬߆ߒหᤨߦᭂ㒢ⅣႺ↢‛ࠍ฽߼ߚ࿾⃿↢๮ߪ‫ߘ߅ޔ‬ࠄߊቝቮ↢๮ߣߒߡߪ଀ᄖ⊛ߦ❥ᩕ࡮ᜬ⛯ߒߡ޿ࠆᚑഞ଀ߩ৻ߟߢ޽ࠅ‫߇ࠇߘޔ‬ቝቮߦ߅޿ߡᭂ߼ߡᏗ
ߥ․ᱶߥ↢๮ࠪࠬ࠹ࡓࠍၮ⋚ߣߒߡ޿ࠆ‫ߣޔ‬⠨߃ࠆᔅὼᕈߪᭂ߼ߡૐ޿‫ࠈߒ߻ޕ‬࿾⃿↢๮ߩሽ࿷น⢻᧦ઙߪ‫ޔ‬ቝቮߦ߅ߌࠆ↢๮ሽ࿷᧦ઙߩၮ⋚ߣߥࠆߣ⠨߃ࠆᣇ߇㆝߆ߦℂߦ߆ߥߞߡ޿ࠆ‫ޕ‬ ߎߩࠃ߁ߥⷰὐ߆ࠄ‫ߪߢ࡯ࡘࡆ࡟࠾ࡒᧄޔ‬ᭂ㒢ⅣႺ߿ߘߎߦ↢ᕷߔࠆ↢‛ߩ⎇ⓥࠍㅢߓߡ⷗߃ߡ߈ߚ
࿾⃿ߩ↢๮ሽ࿷᧦ઙߩႺ⇇ߦߟ޿ߡ᭎⺑ߒߚ޿‫ߎޕ‬ࠇ߹ߢ࿾⃿߿ቝቮߦ߅ߌࠆ↢๮ሽ࿷㗔ၞࠍតࠈ߁ߣ
ߔࠆ⎇ⓥߪ‫ޔ‬ੑߟߩ஥㕙߆ࠄㅴ߼ࠄࠇߡ߈ߚ‫ߟ৻ޕ‬ߪ࿾⃿ߦ߅ߌࠆ↢๮ᵴേߩ㒢⇇(limits of life)ࠍ⷗ᭂ
߼‫↢ߩߘޔ‬๮↢⢒ⅣႺߩ㒢⇇(limits of biosphere)߆ࠄ
↢๮ሽ࿷น⢻ⅣႺࠍតࠈ߁ߣߔࠆࠕࡊࡠ࡯࠴ߢ޽ࠅ‫ޔ‬߽߁৻ߟߪ‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮ሽ࿷᧦ઙ(habitability)ߣ޿߁߽ߩࠍ‛ℂ࡮ൻቇ⊛ߦⷙቯߒ‫‛ߩߘޔ‬ℂ࡮ൻ
ቇ᧦ઙࠍḩߚߔⅣႺࠍតࠈ߁ߣߔࠆࠕࡊࡠ࡯࠴ߢ޽
ࠆ‫ߩߎޕ‬ੑߟߩࠕࡊࡠ࡯࠴ߦࠃߞߡ⷗߃ߡ߈ߚ࿾⃿↢๮ሽ࿷᧦ઙߩᦨᣂ௝ߦㄼߞߡߺࠃ߁‫ޕ‬࿾⃿ߦ߅ߌࠆ↢๮ሽ࿷㒢⇇ ࿾⃿ߦ߅ߌࠆ↢๮ᵴേߪᭂ߼ߡᄙߊߩⷐ࿃ߩ⋧੕૞↪ߦࠃߞߡⷙቯߐࠇࠆ߽ߩߢߪ޽ࠆ߇‫⃻ޔ‬ታߩ࿾
⃿ⅣႺߢߪߔߢߦᄙߊߩⷐ࿃߇ߔߢߦ↢๮ሽ࿷น⢻97᧦ઙߩ▸࿐ౝߦ޽ࠅ‫↢ߩߘޔ‬๮ᵴേ߿↢⢒ⅣႺߩ㒢⇇ࠍⷙቯߔࠆⷐ࿃߇ήᢙߦ޽ࠆߣ޿߁ߎߣߪߥ޿ߛ
ࠈ߁‫ޕ‬ታ㓙ᚒ‫⋥߇ޘ‬ធⷰኤߢ߈ࠆ࿾⃿⴫ጀⅣႺ㧔ᷓᶏ߿࿾Ზ਄ㇱ߽฽߼ߡ㧕ߦ߅޿ߡߪ‫↢ߊోࠈߒ߻ޔ‬๮ᵴേ߇ߥ޿‫↢߿ߩ߽ߩߘ‛↢ߪ޿ࠆ޽ޔ‬๮ᵴേߩ∥〔‛⾰߇฽߹ࠇߥ޿ࠃ߁ߥ↢๮㕖ሽ࿷ⅣႺࠍតߔ
߶߁߇㆔߆ߦ㔍ߒ޿‫ޕ‬࿾⃿ᄖ↢๮ሽ࿷ߩᔅⷐᦨૐ೨
ឭ᧦ઙ߇‫ޟߣޠ࡯ࠡ࡞ࡀࠛ↱⥄ޟ‬ᶧ૕ߩ᳓‫ߩޠ‬ሽ࿷ߢ޽ࠆߣ઒ቯߒߚ႐ว‫߽ࠄߜߤޔ‬࿾⃿⴫ጀⅣႺߦ߅޿ߡ⼾ንߦሽ࿷ߔࠆ᧦ઙߢ޽ࠆ‫ࠇߘޕ‬એᄖߩᭂ߼ߡ
᳿ቯ⊛ߥ᧦ઙߣߒߡߪ‫ޔ‬᷷ᐲ‫ޔ‬࿶ജߣ޿ߞߚ‛ℂ⊛
᧦ઙ߿ pH‫ޔ‬ႮỚᐲߣ޿ߞߚൻቇ⊛᧦ઙࠍ᜼ߍࠆߎߣ
߇ߢ߈ࠆ‫ޕ‬࿾⃿ߦ߅ߌࠆ↢๮ᵴേߩ㒢⇇ࠍㅀߴࠆ೨
ߦ‫↢ߕ߹ޔ‬๮ߦߣߞߡ೑↪น⢻ߥ‫ޟ‬ᶧ૕ߩ᳓‫߇ޠ‬ሽ࿷ߒ߁ࠆᭂ㒢⊛ߥ‛ℂ࡮ൻቇ᧦ઙⅣႺࠍ⠨߃ߡߺࠆ‫ޕ‬ ⴫ 1 ߦߪ‫ޔ‬࿾⃿ߦ߅޿ߡߎࠇ߹ߢߦ⏕⹺ߐࠇߚᶧ
૕ߩ᳓߇↢‛ߦߣߞߡ೑↪น⢻ߥⅣႺߦ߅ߌࠆ᷷ᐲ‫ޔ‬࿶ജ‫ޔ‬pH‫ޔ‬ႮỚᐲߩ᧦ઙߩᭂ㒢ⅣႺ଀߇␜ߐࠇߡ޿
ࠆ 36)‫ߦߢ߹ࠇߎޕ‬⍮ࠄࠇࠆ࿾⃿ⅣႺⅣႺߦ߅ߌࠆᶧ
૕ߩ᳓ߩᦨ㜞᷷ᐲߪ 407 C ߢ޽ࠅ‫ޔ‬ᄢ⷏ᵗਛᄩᶏᎨߦ⷗ߟ߆ߞߚᷓᶏᾲ᳓ྃ಴ሹ߆ࠄྃ಴ߔࠆᾲ᳓ߩ᷷
ᐲߢ޽ࠆ‫ߩߎޕ‬᷷ᐲߪᶏ᳓ߩ⥃⇇ὐ᷷ᐲ(407 C)ߢ޽
ࠅ 3)‫ޔ‬ᶧ૕ߩ᳓ߩℂ⺰⊛ᦨ㜞᷷ᐲߣ⸒߃ࠆ‫ޕ‬ૐ᷷஥ߪ᳖ὐਅߢߪሽ࿷ߔࠆ᳓ߩ߶ߣࠎߤ߇࿕૕ߦߥߞߡ
ߒ߹߁ߩߢᶧ૕ߩ᳓ߩ೑↪߇ᭂ߼ߡ࿎㔍ߦߥࠆ߇‫ޔ‬᳖ਛߦ߅޿ߡ߽ࠊߕ߆ߥᶧ૕ߩ᳓ߩ೑↪น⢻ᕈ߇⠨߃ࠄࠇߡ߅ࠅ‫ޔ‬ᭂ㒢ⅣႺߣߒߡߪ⸥㍳߇ᱷߞߡ޿ࠆ
᷹ቯᦨૐ᳇᷷ࠍ᜼ߍߚ‫ޕ‬࿶ജߦߟ޿ߡ⏕⹺ߐࠇߚ⥄ὼⅣႺߩᦨᄢ㒢⇇ߪ‫࠽ࠕ࡝ࡑޔ‬ᶏḴ࠴ࡖ࡟ࡦࠫࡖ࡯
ᶏᷗߩၸⓍ‛ⅣႺߩ 110 MPa ߢ޽ࠆ 16)‫ޕ‬ૐ࿶஥ߪ‫ޔ‬ቝቮߣ࿾⃿ߩᄢ᳇࿤ߩႺ⇇ߢ޽ࠆᚑጀ࿤਄㒢ߩ᳇࿶ࠍ␜ߔ‫ޕ‬pH ߦߟ޿ߡߪ‫࠽ࠕ࡝ࡑޔ‬೨ᒐၞධ࠴ࡖࡕࡠ
ᶏጊߩⰬ⚉ጤᵹ૕߇ pH 12.5 ࠍ␜ߒ‫ߩߘޔ‬ᒝࠕ࡞ࠞ࡝ᕈᵹ૕߇ᭂ߼ߡᄢ߈ߥᶏᐩਅⓨ㑆ࠍභ߼ߡ޿ࠆߎ
ߣ߇⍮ࠄࠇࠆ 32)‫ޕ‬㒽ၞߢߪ‫ޔ‬ർ࡛࡞࠳ࡦߩࡑࠞ࡝ࡦ
ߦಽᏓߔࠆ㜞᷷ᄌᚑ૞↪ࠍฃߌߚࡆ࠴ࡘ࡯ࡔࡦ⾰ᵆἯጤ࡮⍹Ἧጤ࿾Ꮺߦ߅޿ߡ‫ޔ‬pH 12.9 ߩᄤὼߩ࠮ࡔࡦ
࠻࿾ਅ᳓ߩḝ಴߇⹺߼ࠄࠇߡ߅ࠅ‫ޔ‬࿾ਅߢᲧセ⊛ᄢ߈ߥⓨ㑆ࠍᒻᚑߒߡ޿ࠆߎߣ߇⍮ࠄࠇࠆ 27)‫㉄ޕ‬ᕈ஥ߢߪ‫ޔ‬਎⇇ฦ࿾ߢ pH 0 ߩ᷷ᴰ߇⍮ࠄࠇߡ޿ࠆ 34)ߛߌߢߥߊ‫ࠕ࠾࡞ࠜࡈ࡝ࠞߩࠞ࡝ࡔࠕޔ‬Ꮊߩ࿾ਅ㋶ጊ〔ߩ㋶⣂࿾ਅឃ᳓߇ pH -3.6 ߣ޿߁ࡑࠗ࠽ࠬ୯ࠍ␜
ߔߎߣ߇⍮ࠄࠇߡ޿ࠆ 22, 23)‫ޕ‬ႮỚᐲߢߪ‫ޔ‬㘻๺ႮಽỚᐲࠍ⿥߃ߚႮḓ߇਎⇇ਛߦሽ࿷ߒ‫ޔ‬Ⴎࠍ߶߷ోߊ
฽߹ߥ޿⚐᳓ⅣႺ߽޽ࠅ߰ࠇߡሽ࿷ߔࠆ‫ޔઁߩߘޕ‬⴫ 1 ߦߪ␜ߒߡ޿ߥ޿߇᭽‫‛ߥޘ‬ℂ࡮ൻቇ᧦ઙߢߩ
ᭂ㒢ⅣႺ߇⠨߃ࠄࠇࠆ‫ޕ‬

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)⴫ 1 ࿾⃿ߦ߅ߌࠆᶧ૕ߩ᳓߇ሽ࿷ߒ߁ࠆᭂ㒢ⅣႺߣ↢๮ߩሽ࿷߇ᬌ಴ߐࠇ߁ࠆᭂ㒢ⅣႺ
‛ℂ䊶ൻቇⷐ࿃

ᶧ૕䈱᳓䈏ሽ࿷䈚䈉䉎ᭂ㒢ⅣႺ↢๮䈱ሽ࿷䈏ᬌ಴䈘䉏䈉䉎ᭂ㒢ⅣႺૐ᷷ධᭂ䊗䉴䊃䊷䉪ၮ࿾-89 C䈱᳖

᳖ᐥ䈱᳖ਛ

㜞᷷

407 C䈱ᄢ⷏ᵗਛᄩᶏᎨᷓᶏᾲ᳓

365 C䈱ਛᄩ䉟䊮䊄ᵗᶏᎨᷓᶏᾲ᳓

ૐ࿶ᚑጀ࿤䈱᳖(<0.6 KPa)

58 km਄ⓨ䈱ᚑጀ࿤㜞࿶㉄ᕈ䊙䊥䉝䊅ᶏḴ䉼䊞䊧䊮䉳䊞䊷ᶏᷗ䈱ᶏᐩ(110 MPa)䉦䊥䊐䉤䊦䊆䉝Ꮊ࿾ਅ㋶ጊᑄ᳓(pH -3.6)

䊙䊥䉝䊅ᶏḴ䉼䊞䊧䊮䉳䊞䊷ᶏᷗ䈱ᶏᐩ(110 MPa)䉦䊥䊐䉤䊦䊆䉝Ꮊ࿾ਅ㋶ጊᑄ᳓(pH 0)䉝䊦䉦䊥ᕈർ䊣䊦䉻䊮䊙䉦䊥䊮ᄤὼ䉶䊜䊮䊃࿾ਅ᳓(pH 12.9)

ർ䊣䊦䉻䊮䊙䉦䊥䊮ᄤὼ䉶䊜䊮䊃࿾ਅ᳓(pH 12.7)

㜞ႮỚᐲ

Ⴎḓ䉇ጤႮ

Ⴎḓ䉇ጤႮ

ߢ‫ޔ‬ૐ᷷஥ߩ↢ሽ㒢⇇᷷ᐲߪ᳓ߩ᦭ήߦ㑐ࠊࠄߕน⢻ߥ㒢ࠅૐ᷷ߢ޽ࠇ߫⦟޿‫ޕ‬ ↢‛ߩᦨ㜞↢⢒࿶ജߪ‫࠽ࠕ࡝ࡑޔ‬ᶏḴ߆ࠄಽ㔌ߐ
ࠇߚ Colwellia sp. MT41 ᩣߣ޿߁ࡃࠢ࠹࡝ࠕߩ᦭ߔ
ࠆ 130 MPa ߣ⸒߁⸥㍳߇޽ࠆ 42)‫ޕ‬㜞࿶ਅߢߩ↢‛ߩ↢ሽߦߟ޿ߡߪ‫ࡓ࡜ࠣޔ‬㓁ᕈࡃࠢ࠹࡝ࠕߩᒻᚑߔࠆ
⢩ሶߦߟ޿ߡ⦟ߊ⎇ⓥߐࠇߡ߅ࠅ‫ޔ‬ታ᷹ߢߪ 120 C‫ޔ‬
1.4 GPa ߩ᳓ṁᶧਛߢᢙಽ㑆ಣℂߒߡ߽‫ޔ‬ቢోߦᱫṌߒߥ޿଀߇⍮ࠄࠇߡ޿ࠆ 18)‫ߩߎޕ‬ታ᷹୯߆ࠄ੍ᗐߐ
ࠇࠆ᳓ṁᶧਛߩℂ⺰㒢⇇↢ሽ࿶ജߪ 120 C ߢ 2 GPaߢ޽ࠅ‫ߪࠇߎޔ‬࿾⃿ߢߪ 150km એ਄ߩᷓᶏ߿ 50km
એ਄ߩᷓㇱߩ࿾ਅ᳓ߦ⋧ᒰߔࠆ࿶ജߢ޽ࠅ‫⃻ޔ‬ታߘߩࠃ߁ߥ᳓࿤ߪሽ࿷ߒߥ޿‫ߚ߹ޕ‬⢩ሶߢߪߥߊᓸ↢‛ᩕ㙃⚦⢩ߢ߽‫ޔ‬700 MPa ߢߪ↢ሽߢ߈ࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ 17)‫ߦࠄߐޕ‬᳓ṁᶧਛߢߪߥߊੇ῎⁁ᘒߩ↢‛ߩ࿶ജߦኻߔࠆ↢ሽ⢻ജߪࠃࠅ㜞ߊ‫ࠪࡓࡑࠢޔ‬ߩ৻⒳ߢ޽ࠆ࡛ࠦ࠭࠽ࠢࡑࡓࠪߪ‫⁁⌁ੇޔ‬ᘒߢߪ 7.5
GPa ߩ࿶ജਅߢ 13 ᤨ㑆ಣℂߒߡ߽ᱫṌߖߕ 24)‫ࠃޔ‬ࠅࠨࠗ࠭ߩዊߐ޿ࠦࠤ㘃ߩ⢩ሶߪߘࠇએ਄ߩ㜞࿶⠴ᕈࠍ␜ߔࠄߒ޿‫ߊࠄߘ߅ޕ‬ᓸ↢‛ߩੇ῎⩶૕߿ੇ῎
⢩ሶߩ㜞࿶⠴ᕈߪ‫ߦࠄߐޔ‬ᩴ㆑޿ߦ㜞޿ߎߣ߇੍ᗐ
ߐࠇࠆ߇‫ޔ‬ᱷᔨߥ߇ࠄ⎇ⓥ଀ߪႎ๔ߐࠇߡ޿ߥ޿‫ޕ‬ૐ࿶᧦ઙߦߟ޿ߡߪ‫ޔ‬ૐ᷷ߣหߓࠃ߁ߦ೑↪ߒᓧࠆᶧ૕ߩ᳓ߩሽ࿷㊂߇↢⢒ߩᓞㅦߦߥࠅ‫ޔ‬ㅒߦᶧ૕ߩ
᳓߇ૐ࿶ߦࠃߞߡ⫳᳇ߣߥࠆߎߣߦࠃߞߡ‫ࠁࠊ޿ޔ‬ࠆੇ῎⁁ᘒߦㄭߠ޿ߡ޿ߊߚ߼㐳ᦼ↢ሽߦߪ᦭೑ߦ
௛ߊߎߣ߇⠨߃ࠄࠇࠆ‫ޕ‬ታ㓙࿾਄߆ࠄ 58 km ਄ⓨߩᚑጀ࿤߆ࠄ↢߈ߚᓸ↢‛߇ណขߐࠇߡ߅ࠅ 12)‫ޔ‬ᓸ↢‛⚦⢩߿ᓸ↢‛߅ࠃ߮ᬀ‛ߩ⢩ሶߩ↢ሽߪᒰὼߣߒ
ߡ‫࡞࠱ࡠࠕࠛޔ‬ਛߢߩ↢⢒ߩน⢻ᕈ߽චಽ⠨߃ࠄࠇࠆ‫ޕ‬ ↢‛ߩ↢⢒ pH 㒢⇇ߦߟ޿ߡߪ‫㉄ޔ‬ᕈ஥ߢߪ pH 0ߢ↢⢒ߢ߈ࠆࠕ࡯ࠠࠕ߇⍮ࠄࠇߡ޿ࠆ 8, 34)‫ ߚ߹ޕ‬pH
0 ㄭߊߩ᧦ઙߢ↢⢒ߢ߈‫ޔ‬pH0 ߢ↢ሽน⢻ߥ⌀ᩭ↢‛߽⷗ߟ߆ߞߡ޿ࠆ 7, 31)‫࡝ࠞ࡞ࠕޕ‬ᕈ஥ߢߪ‫ޔ‬ධࠕ
ࡈ࡝ࠞ㊄㋶ߩ࿾ਅ᳓ߩࠦࡦࠢ࡝࡯࠻⾂᳓ᮏ(pH 10.5)߆ࠄಽ㔌ߐࠇߚࠣ࡜ࡓ㓁ᕈࡃࠢ࠹࡝ࠕ Alkaliphilustransvaalensis ߇ pH 12.4 ߹ߢ↢⢒ߔࠆߎߣ߇ߢ߈‫ޔ‬ߎࠇ߇ಽ㔌⩶ᩣߦࠃࠆ↢⢒ pH ࠕ࡞ࠞ࡝ᕈ㒢⇇ߢ޽ࠆ 37)‫৻ޕ‬ᣇ‫ޔ‬ᣢߦ⚫੺ߒߚർ࡛࡞࠳ࡦߩࡑࠞ࡝ࡦߦ
ಽᏓߔࠆ㜞᷷ᄌᚑ૞↪ࠍฃߌߚࡆ࠴ࡘ࡯ࡔࡦ⾰ᵆἯጤ࡮⍹Ἧጤ࿾Ꮺߩ pH 12.7 ࠍ␜ߔ࿾ਅ᳓ߦߪ‫ᦨޔ‬ᄢ
3.3 x 105 cells/ml ߩᓸ↢‛⚦⢩߇ⷰኤߐࠇߡ޿ࠆ 27)‫ޕ‬ ᰴߦ‫ߩࠄࠇߎޔ‬ᶧ૕ߩ᳓߇೑↪น⢻ߥᭂ㒢ⅣႺߩ
‛ℂ࡮ൻቇ᧦ઙ㒢⇇ߦኻߔࠆ↢‛ߩ↢⢒㒢⇇ߣ↢ሽ㒢⇇ߩᐢ߇ࠅߟ޿ߡ⠨߃ࠆ(࿑ 1)36)‫↢ޕ‬๮ሽ࿷น⢻᧦ઙߣߪ‫⃻߇‛↢ߪߢ⟵⁜ޔ‬ሽࡃࠗࠝࡑࠬࠍჇᄢߐߖ
ࠆ(growth)߽ߒߊߪ⛽ᜬ(maintenance)ߢ߈ࠆ᧦ઙߢ
޽ࠅ‫ߩߘޔ‬㒢⇇᧦ઙ߇↢⢒㒢⇇ߢ޽ࠆ‫৻ޕ‬ᣇ‫ޔ‬ᐢ⟵ߩ↢๮ሽ࿷น⢻᧦ઙߦߪ‫↢ޔ‬⢒㒢⇇ߛߌߢߥߊ‫↢ޔ‬‛ߩ⃻ሽࡃࠗࠝࡑࠬ߿୘૕(⚦⢩)߇৻ቯᤨ㑆ߦቢోߦᱫṌߒߥ޿᧦ઙ‫↢ޔ‬ሽ㒢⇇߹ߢ߇฽߹ࠇߡ޿ࠆ႐
ว߇ᄙ޿‫ޕ‬ ⃻࿷߹ߢߦ⍮ࠄࠇࠆ↢‛ߩᦨ㜞↢⢒᷷ᐲߪ‫ࡦࠗޔ‬࠼ᵗߩᷓᶏᐩᾲ᳓ᵴേၞߩ㜞᷷ᾲ᳓ਛ(365 C)ߢ↢ᚑߐࠇߚ㋶‛߆ࠄಽ㔌ߐࠇߚࠕ࡯ࠠࠕ Methanopyrus
kandleri 116 ᩣߩ㜞᳓࿶ਅߢߩᦨ㜞Ⴧᱺ᷷ᐲ 122 Cߢ޽ࠅ‫ ߪ⩶ᧄߚ߹ޔ‬108 cells/ml ߩ⚦⢩ኒᐲࠍ᦭ߔࠆ
ၭ࿾ਛߢߪ‫ޔ‬130 C ߢ 3 ᤨ㑆ಣℂߒߡ߽ቢోᱫṌߔࠆߎߣߪߥ޿ 38)‫⃻ޔߒߛߚޕ‬႐ⅣႺߩ 365 C ߩᾲ᳓ߦჇᱺน⢻ߥᓸ↢‛߇ᬌ಴ߐࠇߡ޿ࠆߩߢ 38)‫ޔ‬᷷ᐲߦߟ޿ߡߪ᷷ᐲ࡮࿶ജએᄖߩ⥄ὼⅣႺߩ᧦ઙࠍౣ⃻ߒߡ⹏ଔߔࠆᔅⷐ߇޽ࠆ 36)‫ޕ‬ᶧ૕ߩ᳓߇߶ߣࠎߤሽ
࿷ߒߥ޿ੇ῎᧦ઙߢߪ‫ޔ‬㜞᷷ߢߩ↢ሽน⢻ᕈߪࠃࠅჇᄢߔࠆߣ⠨߃ࠄࠇߡ޿ࠆ‫ޕ‬㘩ຠⴡ↢ቇߩಽ㊁ߢߪ‫ޔ‬300 C‫ޔ‬30 ಽߩੇᾲṌ⩶ߢߪࠣ࡜ࡓ㓁ᕈࡃࠢ࠹࡝ࠕߩੇ῎⢩ሶࠍቢోߦᱫṌߐߖࠆߎߣ߇ߢ߈ߥ޿⃻⽎߇ࠃߊ⍮ࠄࠇߡ߅ࠅ‫⁁⌁ੇޔ‬ᘒߩ✭ᱠേ‛㐷ߩࠢࡑ
ࡓࠪߩ৻⒳ߪ‫ޔ‬151 C ߢ 35 ಽߩੇᾲಣℂᓟ߽ᱫṌߒ
ߥ޿ 30)‫ߒ߆ߒޕ‬ᱷᔨߥ߇ࠄ‫ޔ‬ఝࠇߚ㜞᷷⠴ᕈࠍ␜ߔ
⿥ᅢᾲ⩶ߩੇ῎⩶૕߇‫޿ࠄߋࠇߤޔ‬㜞᷷࡮ੇ῎⁁ᘒߢߩ↢ሽ⢻ജࠍ᦭ߔࠆߩ߆ߦߟ޿ߡߪ߶ߣࠎߤోߊ
⎇ⓥ଀߇ߥ޿‫ޕ‬ ෻ኻߦ↢‛ߩᦨૐ↢⢒᷷ᐲߪ‫ޔ‬ታ㓙ߩჇᱺታ㛎ߢ᷹ቯߐࠇߚ⸥㍳ߣߒߡߪࡃࠢ࠹࡝ࠕ Psychromonas
ingrahamii ߩᦨૐჇᱺ᷷ᐲ-12 C6)‫ޔ‬Ⴧᱺㅦᐲቯᢙߣ᷷ ᐲ ߩ ℂ ⺰ ⊛ ੍ ᗐ ߆ ࠄ ࡃ ࠢ ࠹ ࡝ ࠕ Shewanella
gelidimarina ߩផቯᦨૐჇᱺ᷷ᐲ-27 C5)‫߇ޔ‬⍮ࠄࠇߡ޿ࠆ‫ߚ߹ޕ‬ᓸ↢‛ߩ๭ๆᵴᕈߪ‫ޔ‬-33 C ߢ߽ᬌ಴ߐࠇߡ޿ࠆ 1)‫ޕ‬ૐ᷷ߢߩ↢ሽ㒢⇇ߦߟ޿ߡߪ‫⥸৻ޔ‬ߦᓸ↢‛ߦ㒢ࠄߕ‫⚦ޔ‬⢩ߪ಄ಓ⁁ᘒߦ޽ࠆߣᭂ߼ߡ
㐳ᦼ㑆↢ሽߔࠆ‫ޕ‬಄ಓ⁁ᘒߦ޽ࠆ⚦⢩ߩ↢ሽࠍ᳿ቯߠߌࠆ࿃ሶߣߒߡ‫ޔ‬᭴ᚑ࠲ࡦࡄࠢ⾰ਛߩࠕࡒࡁ㉄ߩ࡜࠮ࡒൻߣ᭴ᚑ DNA ߩ⣕ࡊ࡝ࡦൻ߇ᜰ៰ߐࠇߡ޿ࠆ 20)‫ߩࠄࠇߎޕ‬ൻቇ෻ᔕߪ᳓ಽᵴᕈߩૐਅߦᓥ޿‫ޔ‬޽ࠆ޿ߪ᷷ᐲߩૐਅߦᓥ޿ߘߩ෻ᔕߪᛥ೙ߐࠇࠆߩ
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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)෼ߐࠇߚᦨ߽ฎ޿ᓸ↢‛ߩႎ๔ߪ‫ޔ‬2.5 ంᐕ೨ߩBacillus ዻࡃࠢ࠹࡝ࠕ 40)‫ޔ‬ฎઍ DNA ߣߒߡߩႎ๔ߪ‫ޔ‬
4.2 ంᐕ೨ߩ㜞ᐲᅢႮࠕ࡯ࠠࠕߩㆮવሶ 25)߇޽ࠅ‫ޔ‬޿ߕࠇ߽଻ሽߩ⦟޿࿾ਅጤႮ߆ࠄ࿁෼ߐࠇߡ޿ࠆ‫ޕ‬ߚߛߒ‫ߩࠄࠇߘޔ‬ႎ๔ߪ੐ታߣߒߡᐢߊฃߌ޿ࠇࠄ
ࠇߡ޿ࠆࠊߌߢߪߥ޿‫ޕ‬ એ਄‫ޔ‬ฦ‫ߩޘ‬ਥⷐߥ‛ℂ࡮ൻቇ᧦ઙߦ߅ߌࠆ↢๮ߩ↢⢒㒢⇇ߣ↢ሽ㒢⇇ߦߟ޿ߡ᭎⺑ߒߚ‫ࠄࠇߎޕ‬࿾⃿↢๮ߩ↢⢒߿↢ሽߦ߅ߌࠆන⁛ߩ‛ℂ࡮ൻቇⅣႺ᧦ઙߩ㒢⇇ߪ‫ᤨ⃻ߢ߹ߊ޽ޔ‬ὐߢߩ೔㆐ὐߢ޽ࠅ‫ޔ‬੹ᓟ߽⎇ⓥߩㅴዷߦࠃߞߡ᜛ᒛߐࠇߡࠁߊߎߣߪ㑆㆑޿ߥ޿‫ߩࠄࠇߎޕ‬㒢⇇୯ߪ‫ޔ‬ዋߥߊߣ߽ฦ‛ℂ࡮
ൻቇ᧦ઙ߇ߎߩ▸࿐ౝߦ޽ࠇ߫‫ޔ‬࿾⃿↢๮߇↢⢒޽ࠆ޿ߪሽ࿷ߒᓧࠆߣ޿߁ᦨዊߩ▸࿐ࠍ␜ߔ߽ߩߢ޽
ࠆ‫↢ߩߎޕ‬๮ሽ࿷᧦ઙ▸࿐ࠍ࿾⃿ߦ߅޿ߡ⏕⹺ߐࠇߚᭂ㒢ⅣႺߣᲧセߒߚ႐ว(࿑ 1)‫ޔ‬᷷ᐲࠍ㒰ߊ߶ߣࠎߤߔߴߡߩ᧦ઙߪ‫↢ߦߢߔޔ‬⢒น⢻ߥ▸࿐ߦ޽ࠆߎ
ߣ߇ࠊ߆ࠆߢ޽ࠈ߁‫ޔ߽ࠄ߆ߣߎߩߎޕ‬㜞᷷ⅣႺࠍ
㒰ߌ߫࿾⃿ߦ߅޿ߡ↢๮߇ሽ࿷ߢ߈ߥ޿ⅣႺ߇߶ߣࠎߤߥ޿ߣ޿߁ߎߣ߽ℂ⸃ߢ߈ࠆ‫ࠄࠇߎޔߒߛߚޕ‬ߩ⠨ኤߪ޽ߊ߹ߢ‫ޔ‬න⁛ߩ᧦ઙߦ㒢ቯߒߚ႐วߢ޽
ࠅ‫ޔ‬᭽‫᧦ߥޘ‬ઙ߇ⶄวߒߚ႐ว‫ߩࠇߙࠇߘޔ‬㒢⇇୯ߪ⋧ਸ਼⊛ߦᓇ㗀ࠍฃߌࠆ⚵ߺวࠊߖ߽޽ࠆ‫ޕ‬ታ㓙ߩⶄวᭂ㒢ⅣႺ᧦ઙߢߪ‫ޔ‬න⁛ߩ᧦ઙࠃࠅ߽㒢⇇୯߇߆ߥࠅૐߊߥࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ 21)‫ޕ‬ߎߩ਎⇇ᦨ㜞 pH ࠍ␜ߔ࿾ਅ᳓ߦ᦭ᯏ㉄ࠍᷝടߒ⃻႐ၭ㙃ߒߚߣߎࠈ‫ޔ‬ᓸ↢‛ߩࡃࠗࠝࡑࠬߩჇട߇ⷰ
ኤߐࠇߡ޿ࠆ 27)‫ߩᤨߩߘޕ‬࿾ਅ᳓ߩ pH ߪ 12.5 ߢ޽ࠅ‫ޔ‬ᓸ↢‛⟲㓸ߩఝභ⒳ߪ Pseudomonas ዻࡃࠢ࠹࡝
ࠕߢ޽ߞߚߣႎ๔ߐࠇߡ޿ࠆ 27)‫⚿ߩߎߒ߆ߒޕ‬ᨐࠍ↢‛ߩ↢⢒ pH ࠕ࡞ࠞ࡝ᕈ㒢⇇ߣ⠨߃ࠆߦߪ߹ߛ⸽
᜚߇ਇචಽߢ޽ࠆ‫ ߚ߹ޕ‬Alkaliphilus transvaalensisߥߤߩᅢࠕ࡞ࠞ࡝ࡃࠢ࠹࡝ࠕߩ↢ሽߦ෸߷ߔ pH ߩᓇ㗀ߪ⺞ߴࠄࠇߚ⎇ⓥ଀ߪߥ޿߇‫↢ޔ‬⢒㒢⇇ࠍ⿥߃ࠆ pH ߢ↢ሽน⢻ߢ޽ࠆߎߣߪኈᤃߦᗐ௝ߢ߈‫߆ޔ‬ߟ Alkaliphilus transvaalensis ߪ⢩ሶࠍᒻᚑߔࠆߎߣ
߇ߢ߈ࠆ‫ࡓ࡜ࠣޕ‬㓁ᕈࡃࠢ࠹࡝ࠕߩᒻᚑߔࠆ⢩ሶߩኻൻቇ࿃ሶ⠴ᕈߪ‫ޔ‬ᾲ߿࿶ജߣ޿ߞߚኻ‛ℂ࿃ሶ⠴
ᕈࠍ㆔߆ߦ਄࿁ࠆߎߣ߇⍮ࠄࠇߡ޿ࠆߩߢ‫ࠄߘ߅ޔ‬ߊ Alkaliphilus transvaalensis ߩ⢩ሶߩ pH ⠴ᕈߪ pH14ߢ޽ߞߡ߽ߔߊߥߊߣ߽ᢙᣣ߆ࠄᢙࡩ᦬ߪኈᤃߦ↢ሽߢ߈ࠆߣផ᷹ߐࠇࠆ‫ޕ‬ ↢‛ߩ↢⢒߿↢ሽߦ߅ߌࠆႮỚᐲߩ㒢⇇᧦ઙߣߒߡߪ‫ޔ‬ᩕ㙃ߣߒߡߩήᯏ࡮᦭ᯏ‛Ⴎࠍ㒰෰ߒߥ޿ߣ
޿߁೨ឭߦ┙ߡ߫‫ޔ‬ᄙߊߩ㒽᳓ᕈ↢‛߿ᓸ↢‛ߪႮỚᐲ߶߷ 0 ߢ↢⢒น⢻ߢ޽ࠅ‫ޔ‬㘻๺ႮỚᐲ(NaCl ߢ឵▚ߒߡ⚂ 30 % w/v)ߢ޽ߞߡ߽‫ޔ‬ᄙߊߩ㜞ᐲᅢႮࠕ࡯ࠠࠕߣ޿ߊߟ߆ߩ㜞ᐲᅢႮࡃࠢ࠹࡝ࠕߪ↢⢒น⢻ߢ޽ࠆ‫↢ߚ߹ޕ‬๮ߩ↢ሽߦ㑐ߒߡߪ‫ޔ‬㜞Ⴎಽ᳓߆ࠄ
↢ߓࠆႮߩ⚿᥏(ጤႮ)ߪ‫⥄ޔ‬ὼⅣႺߦሽ࿷ߔࠆᩰᅢ
ߩᓸ↢‛↢ሽⅣႺߣߥࠆ 40)‫ߚ߈↢ޕ‬ᓸ↢‛ߣߒߡ࿁࿑ 1 ࿾⃿ߦ߅ߌࠆᶧ૕ߩ᳓߇೑↪น⢻ߥᭂ㒢ⅣႺߩ‛ℂ࡮ൻቇ᧦ઙ㒢⇇ߣ↢‛ߩ↢⢒㒢⇇ߣ↢ሽ㒢⇇‫(ޕ‬A)ᶧ૕ߩ᳓߇೑↪น⢻ߥᭂ㒢ⅣႺߩ‛ℂ࡮ൻቇ᧦ઙ㒢⇇(⭯޿Ἧ⦡ߩ㗔ၞ)ߦኻߔࠆታ㛎ቶౝߢౣ⃻น⢻ߥᓸ↢‛ߩ↢⢒(Ⴧᱺ)㒢⇇᧦ઙ(Ớ޿Ἧ⦡ߩ㗔ၞ)ߩᐢ߇ࠅ‫(ޕ‬B) ᶧ૕ߩ᳓߇೑↪น⢻ߥᭂ㒢ⅣႺߩ‛ℂ࡮ൻቇ᧦ઙ㒢⇇(⭯޿Ἧ⦡ߩ㗔ၞ)ߦኻߔࠆታ㛎ቶౝߢౣ⃻น⢻ߥ᳓ਛߦ߅ߌࠆᓸ↢‛ߩ↢ሽ㒢⇇᧦ઙ(Ớ޿Ἧ⦡ߩ㗔ၞ)ߩᐢ
߇ࠅ‫ޕ‬

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ᭂ 㒢 ⅣႺ ↢‛ ߩ ߩࠛ ࡀ࡞ ࠡ ࡯⺰ ⊛↢ ๮ ሽ࿷ ᧦ઙ(habitability) ߎߎ߹ߢ⚫੺ߒߚฦਥⷐߥ‛ℂ࡮ൻቇ᧦ઙߦ߅ߌ
ࠆ࿾⃿ߦ߅ߌࠆ↢‛ߩ↢⢒㒢⇇ߣ↢ሽ㒢⇇ߪ‫ޔ‬ᭂ߼ߡ㒢ቯߐࠇߚ㗄⋡᧦ઙߣߪ⸒߃‫ޔ‬᣿⏕ߥ↢๮ሽ࿷᧦ઙ㗔ၞࠍ⴫ߔ߽ߩߣ⸒߃ࠆ‫ޔߢ߹ߊ޽ޔ߃޿ߪߣޕ‬‫ᤨ⃻ޟ‬ὐ߹ߢߦ⍮ࠄࠇࠆ࿾⃿↢๮ߩᦨ߽ዊߐߊ⷗Ⓧ߽ࠄࠇߚ↢๮ሽ࿷᧦ઙߩ৻┵‫ߒ߆ߒޕ޿ߥ߉ߔߦޠ‬࿾⃿ߦ߅ߌࠆ↢๮ᵴേߩ㒢⇇߿࿾⃿ᄖ↢๮ߩ↢๮ᵴേࠍ⷗ᭂ߼ࠆߦߪ‫ޔ‬‫ޟ‬࿾⃿↢๮ߩ↢๮ሽ࿷᧦ઙߩ৻┵‫ޠ‬ߛߌߢߥߊ‫ޔ‬ቝቮ౒ㅢේℂߣߒߡ⑼ቇ⊛ߦᬌ⸽น⢻ߥ⏕࿕ߚࠆ᜚ᚲߣߥࠆ᭎ᔨ߇ᔅⷐߢ޽ࠆ‫߆ߥߩߘޕ‬ߢ⠨߃಴ߐࠇߚ᭎ᔨ߇‫ޔ‬habitability(↢๮ሽ࿷᧦ઙ)ߢ޽ࠆ‫ޕ‬
 ߘߒߡߎߩ↢๮ሽ࿷᧦ઙߣ޿߁᭎ᔨࠍೋ߼ߡ౏ᑼ
ൻߒߚߩߪ Hoehler10)ߢ޽ࠆ‫ޕ‬ᓐߩឭ᩺ߔࠆ‫࡞ࡀࠛޟ‬ࠡ࡯⺰ߣߒߡߩ↢๮ሽ࿷᧦ઙ‫߁޿ߣޠ‬᭎ᔨߪ‫ޔ‬ᭂ߼ߡⶄ㔀ߥ♽ߣߒߡߩ↢๮ᵴേࠍ‫ࡦ࠹ࡐ࡯ࠡ࡞ࡀࠛޔ‬ࠪࡖ࡞(㔚࿶)ߣࠛࡀ࡞ࠡ࡯㊂(㔚ജ)ߩੑߟߩⷐ⚛ߛߌߢ೙⚂ߔࠆ߽ߩߢ޽ߞߚ(࿑ 2)‫ޕ‬Hoehler10)ߪ‫ࡀࠛޔ‬࡞ࠡ࡯⺰ߣߒߡߩ↢๮ሽ࿷᧦ઙߪ‫ޔ‬એਅߩ 4 ߟߩߒ߈޿୯ߢ✼ขࠄࠇࠆࠛࡀ࡞ࠡ࡯㗔ၞߢ޽ࠆߣቯ⟵ߒߚ‫ޕ‬(1)↢๮ᵴേߩේേജߦߥࠆ‫ⶄߩ♽ޟ‬㔀ߐ‫⛽ࠍޠ‬ᜬߔࠆ ᦨ ዊ ߩ ࠛ ࡀ ࡞ ࠡ ࡯ ࡐ ࠹ ࡦ ࠪ ࡖ ࡞ (biologicalenergy quantum: BEQ)‫ࠇߎࠅ߹ߟޕ‬એਅߩࠛࡀ࡞
ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞ߢߪ↢๮ᵴേߩߚ߼ߦ೑↪ߢ߈ߥ޿ߣ޿߁㒢⇇୯‫ޕ‬(2)‫ⶄߩ♽ޟ‬㔀ߐ‫⛽߇ޠ‬ᜬߐࠇᓧࠆᦨᄢߩࠛࡀ࡞ࠡ࡯
ࡐ࠹ࡦࠪࡖ࡞‫ࠇߎࠅ߹ߟޕ‬એ਄ߩࠛࡀ࡞ࠡ࡯ࡐ࠹
ࡦࠪࡖ࡞ߢߪ♽߇೙ᓮਇ⢻ߦߥࠆ㒢⇇୯‫ޕ‬(3) ‫ ⶄ ߩ ♽ ޟ‬㔀 ߐ ‫ ⛽ ࠍ ޠ‬ᜬ ߔ ࠆ ᦨ ዊ ߩ ઀ ੐ ㊂(maintenance energy: ME)‫↢ޕ‬๮ᵴേߩ⛽ᜬߦᔅⷐߥᦨዊߩࠛࡀ࡞ࠡ࡯㊂‫ޕ‬(4)‫ⶄߩ♽ޟ‬㔀ߐ‫߇ޠ‬ฃߌ౉ࠇࠆߎߣߩߢ߈ࠆᦨᄢߩ
ࠛࡀ࡞ࠡ࡯㊂‫↢ࠅ߹ߟޕ‬๮ᵴേࠍ᥸⿛ߐߖߥ޿ᦨ
ዊߩࠛࡀ࡞ࠡ࡯㊂‫ޕ‬ ߎߩ 4 ߟߩߒ߈޿୯ߩ߁ߜ‫ࠪࡦ࠹ࡐ࡯ࠡ࡞ࡀࠛޔ‬ࡖ࡞ߣࠛࡀ࡞ࠡ࡯㊂ߦ㑐ߔࠆ਄ᣇߩႺ⇇ߪ‫↢ޔ‬๮ᵴേߩṁᇦߢ޽ࠆ᳓߿ߘߩઁߩน⢻ᕈߩ޽ࠆṁᇦߩሽ࿷⁁ᘒߦᄢ߈ߊଐሽߒ‫↢ߩߘޔߚ߹ޔ‬๮ᵴേࠍᡰ߃
ࠆ♽⥄૕ߩ᭴ᚑ߿᭴ㅧ߇᣿⏕ߢߥ޿ߣߘߩ㒢⇇ࠍᱜ⏕ߦ⷗ᭂ߼ࠆߎߣ߇㔍ߒ޿߇‫ޔ‬࿾⃿ߩ↢๮ߦߟ޿ߡ
⸒߃߫‫ޔ‬਄⸥ߩᦨ㜞↢⢒᷷ᐲ߿᡼኿✢↢ሽ⠴ᕈ㒢⇇߿⚡ᄖ✢↢ሽ⠴ᕈ㒢⇇ߩࠃ߁ߥ‫↢ޟ‬๮ᵴേࠍᡰ߃ࠆ
♽‫ߩޠ‬቟ቯᕈߩ㒢⇇ߣᝒ߃ࠆ੐߇ߢ߈ࠆ‫ޕ‬ ߻ߒࠈ↢๮ᵴേߩ⛽ᜬߦᔅⷐߥᦨዊߩࠛࡀ࡞ࠡ࡯
ࡐ࠹ࡦࠪࡖ࡞߿ࠛࡀ࡞ࠡ࡯㊂ߩᣇ߇‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮ሽ࿷㗔ၞࠍ⠨߃ࠆ਄ߢ߽‫ޔ‬࿾⃿ᄖ↢๮តᩏࠍ⠨߃ࠆ਄ߢ߽㊀ⷐߢ޽ࠆ‫↢ߦ․ޕ‬๮ᵴേߩ⛽ᜬߦᔅⷐߥࠛࡀ࡞ࠡ࡯߇ᰳਲߒߡ޿ࠆߣ⠨߃ࠄࠇߡ߈ߚᄢ㒽࿾Ზߩጤ⍹ 27, 35)߿ᷓㇱᶏᐩਅၸⓍ‛ਛ 26)ߦ੍ᗐᄖߩ
ᓸ↢‛ࡃࠗࠝࡑࠬ߇ሽ࿷ߔࠆߣ޿߁⊒⷗߇‫ࠃߩߘޔ‬߁ߥⅣႺߦ߅޿ߡ߽↢๮ሽ࿷᧦ઙ߇ḩߚߐࠇߡ޿ࠆ‫ޔ‬ߔߥࠊߜ BEQ ߿ ME ߇⏕଻ߐࠇߡ޿ࠆ‫߁޿ߣޔ‬੐ታࠍឭ␜ߔࠆߎߣߣߥߞߚ‫ޕ‬࿾⃿ߩᭂ߼ߡ↢๮߇ሽ࿷ߒ㔍޿ߣ⠨߃ࠄࠇߡ߈ߚⅣႺߦ߅޿ߡ↢๮ሽ࿷᧦ઙ
߇ḩߚߐࠇߡ޿ࠆߥࠄ߫‫ޔ‬࿾⃿ᄖᖺᤊ߿ⴡᤊߦ߅޿
ߡ߽↢๮ሽ࿷᧦ઙߪḩߚߐࠇߡ޿ࠆน⢻ᕈߪჇᄢߔ
ࠆ‫ޔ߇ࠇߘޕ‬࿾⃿ߦ߅ߌࠆ࿾⃿↢๮ߩ↢๮ሽ࿷᧦ઙ
ߩ㒢⇇‫↢ࠅ߹ߟޔ‬๮࿤ߩႺ⇇‫ࠍޔ‬តᩏߔࠆ⋡⊛ߩ৻ߟߢ޽ࠆ‫ޕ‬࿑ 2 Hoehler10)ឭ᩺ߔࠆࠛࡀ࡞ࠡ࡯⺰ߣߒߡߩ↢๮ሽ࿷น⢻᧦ઙߩ᭎ᔨ࿑(ේ࿑ࠍᡷᄌ)‫ޕ‬Hoehler10)ߪ‫ࡀࠛޔ‬࡞ࠡ࡯⺰⊛↢๮ሽ࿷น⢻㗔ၞߪ 4 ߟߩߒ߈޿୯ߢ✼ขࠄࠇࠆࠛࡀ࡞ࠡ࡯㗔ၞߢ޽ࠆߣ⚿⺰ߠߌߚ‫ޕ‬100

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2) ߢߪ‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮ߩࠛࡀ࡞ࠡ࡯⺰⊛ሽ࿷᧦ઙ‫ ߦ․ޔ‬BEQ ߿ ME‫ߢ߹ߎߤߪޔ‬᣿ࠄ߆ߦߐࠇߚߩ
ߢ޽ࠈ߁߆㧫BEQ‫↢ࠅ߹ߟޔ‬๮ᵴേࠍ⛽ᜬߔࠆߩߦ
ᔅⷐߥᦨዊ㒢ߩࠛࡀ࡞ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞(ࠡࡉࠬ⥄↱ࠛࡀ࡞ࠡ࡯ᄌൻ)‫↢ߕ߹ޔߪߡ޿ߟߦޔ‬๮ᵴേߩ౒
ㅢࠛࡀ࡞ࠡ࡯⾂⬿‛⾰ߢ޽ࠆࠕ࠺ࡁࠪࡦਃ࡝ࡦ㉄(ATP)߇↢‛⤑ߦ߅޿ߡ↢ᚑߐࠇࠆ㓙ߦᔅⷐߥࠛࡀ
࡞ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞߆ࠄ⷗Ⓧ߽ࠄࠇߚ 33)‫ࠪࡁ࠺ࠕޕ‬ࡦੑ࡝ࡦ㉄(ADP)߆ࠄ ATP ࠍࡊࡠ࠻ࡦ൨㈩㚟േ ATPวᚑ㉂⚛ߢวᚑߔࠆ㓙‫ޔ‬ᔅⷐߥᦨዊ㒢ߩࠡࡉࠬ⥄↱ࠛࡀ࡞ࠡ࡯ᄌൻߪ⚂-60 KJ/molATP ߢ޽ࠅ‫ޔ‬㧝ಽሶߩ ATP ᒰߚࠅ 3 ߟߩࡊࡠ࠻ࡦ(㔚ሶ)߇ᔅⷐߣߐࠇࠆ
ߚ߼‫ޔ‬1 㔚ሶ޽ߚࠅߩࠡࡉࠬ⥄↱ࠛࡀ࡞ࠡ࡯ᄌൻߪ
-20 KJ/mol ߣ੍ᗐߐࠇࠆ‫ޕ‬ታ㓙ߩᓸ↢‛ၭ㙃ታ㛎ߦࠃߞߡ‫ޔ‬ᓸ↢‛ߩჇᱺࠍᡰ߃ࠆᦨዊߢߩࠛࡀ࡞ࠡ࡯
ࡐ࠹ࡦࠪࡖ࡞ࠍ⷗Ⓧ߽ࠆߣ‫ߦ߆⏕ޔ‬-20 KJ/mol ⒟ᐲߢ޽ࠆߎߣ߇᣿ࠄ߆ߦߥߞߚ 33)‫⚿ߩߎޕ‬ᨐߪߟ߹ࠅ‫ޔ‬‫ޟ‬-20 KJ/mol ⒟ᐲ(㔚૏ߢ⸒߃߫-200 mV ⒟ᐲ)એ਄ߩ
ࠡࡉࠬ⥄↱ࠛࡀ࡞ࠡ࡯ᄌൻࠍ↢ߺ಴ߖߥ޿ࠃ߁ߥᾲ
ജቇ⊛㕖ᐔⴧⅣႺ᧦ઙߢߪ↢๮߇ሽ࿷ߢ߈ߥ޿‫ߣޠ‬޿߁ߎߣࠍᗧ๧ߔࠆ‫⃻ߒ߆ߒޕ‬ታߩ࿾⃿ⅣႺ߿ታ㛎ቶౝၭ㙃ⅣႺߢߪ‫ޔ‬-20 KJ/mol એਅߩࠛࡀ࡞ࠡ࡯ࡐ
࠹ࡦࠪࡖ࡞ߒ߆↢ߺ಴ߖߥ޿ࠃ߁ߥ᧦ઙਅߢ߽‫ޔ‬ᓸ↢‛ߪჇᱺ޽ࠆ޿ߪࡃࠗࠝࡑࠬࠍ⛽ᜬߔࠆߎߣ߇ߢ
߈ࠆߎߣ߇᣿ࠄ߆ߦߥߞߚ 14)‫ߥ߁ࠃߩߘޕ‬ᓸ↢‛ߩ↢⢒ߪ౒ᩕ㙃౒↢ߣ޿߁ࡔࠞ࠾࠭ࡓߦଐሽߒߡ߅ࠅ‫ޔ‬౒ᩕ㙃౒↢ᓸ↢‛ၭ㙃ታ㛎♽ߢߩ⚿ᨐ߆ࠄ BEQ ࠍ
⷗Ⓧ߽ߞߚ႐ว‫ᦨޔ‬ዊ୯ߪ-5 KJ/mol ⒟ᐲߣ޿߁୯߇
ᓧࠄࠇߚ 14)‫ ߦࠄߐޕ‬Hoehler11)ߪ‫⥄ޔ‬ὼⅣႺߦ߅ߌࠆ
ࠛࡀ࡞ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞ߩ⷗Ⓧࠅߣᓸ↢‛⟲㓸ߩᵴᕈߦၮߠ߈‫ߘࠃ߅ޔ‬-10 KJ/mol ⒟ᐲ߇ BEQ ߣߥࠆߎߣࠍᜰ៰ߒߡ޿ࠆ‫ࠅ߹ߟޕ‬ᓸ↢‛ታ㛎߿⥄ὼⅣႺਛ
ߩᓸ↢‛⟲㓸ߩᵴᕈ߆ࠄߪ‫ޔ‬ዋߥߊߣ߽-5ޯ-10
KJ/mol ⒟ᐲߩࠛࡀ࡞ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞߇⏕଻ߐࠇࠆࠛࡀ࡞ࠡ࡯ⅣႺߢ޽ࠇ߫‫↢ޔ‬๮ᵴേߩ⛽ᜬ߇น⢻ߢ޽ࠆߎߣ߇ಽ߆ߞߚ‫ޕ‬ BEQ ߪ޽ߊ߹ߢࠛࡀ࡞ࠡ࡯ࡐ࠹ࡦࠪࡖ࡞ߩߒ߈
޿୯ߢ޽ࠅ‫ޔ‬ታ㓙ߦᔅⷐߥࠛࡀ࡞ࠡ࡯㊂ߢߪߥ޿‫ޕ‬↢๮ᵴേࠍ⛽ᜬߔࠆᦨዊߩࠛࡀ࡞ࠡ࡯㊂ߢ޽ࠆߣቯ
⟵ߐࠇࠆ ME ߩᣇ߇‫⃻ޔ‬ታߩⅣႺߢߪࠃࠅᡰ㈩⊛ߢ޽ࠈ߁‫ ߩߎޕ‬ME ߦߟ޿ߡߪ‫ޔ‬ㅦᐲ⺰⊛ 29)෸߮ᾲജ
ቇ⊛ 19)ߥࠕࡊࡠ࡯࠴ߦࠃࠅ⷗Ⓧ߽ࠅ߇ႎ๔ߐࠇߡ޿
ࠆ‫ޕ‬Price and Sower29)ߪ‫ޔ‬ᓸ↢‛⟲㓸ߩ৻ቯࡃࠗࠝࡑ
ࠬᒰߚࠅ(gC)ߩ὇⚛ขࠅㄟߺㅦᐲߣⅣႺ᷷ᐲߩ㑐ଥ߆ࠄ‫ޔ‬࿾⃿ߩ᭽‫↢ߥޘ‬๮ᵴേߩ⛽ᜬ߇࿎㔍ߣ⠨߃ࠄࠇࠆᭂ㒢ⅣႺߦ߅ߌࠆᓸ↢‛⟲㓸ߩሽ࿷⁁ᘒ߇‫ޔ‬
growth(Ⴧᱺ)‫ޔ‬maintenance(⛽ᜬ)‫ޔ‬survival(↢ሽ)ߦಽ
ߌࠄࠇࠆߎߣࠍ⷗಴ߒߚ(Hoehler ߩቯ⟵ߔࠆ ME ߪߎߩ႐ว survival ⁁ᘒߦ⋧ᒰߔࠆ)‫↢ߡߒߘޕ‬๮ᵴേࠍ⛽ᜬߔࠆߚ߼ߩ᧦ઙߪ‫⾰ࠢࡄࡦ࠲ޔ‬ਛߩࠕࠬࡄ࡜
ࠡࡦ㉄ߩ࡜࠮ࡒൻߩ෻ᔕㅦᐲ਄࿁ࠆ⒟ᐲߩઍ⻢ㅦᐲ߇⏕଻ߐࠇࠆᔅⷐ߇޽ࠆߣ޿߁⷗Ⓧ߽ࠅࠍⴕߞߚ‫ޕ‬ߘߩ⚿ᨐߪ‫↢ޔ‬๮ᵴേࠍ⛽ᜬߔࠆߚ߼ߩઍ⻢ㅦᐲߪჇᱺߦᔅⷐߥઍ⻢ㅦᐲߩ 10-3ޯ10-6 ⒟ᐲߦߔ߉ߥ޿
ߎߣࠍ␜ߒߚ(࿑ 3)‫ߩߘޕ‬ઍ⻢ㅦᐲߣߪ‫߫߃଀ޔ‬-40C101ߩ᳖ߦ㐽ߓㄟ߼ࠄࠇߚᓸ↢‛߇↢ሽߔࠆߚ߼ߦߪ‫ޔ‬1ంᐕ㑆ߢ 1 ࿁⒟ᐲߩ୚ടᤨ㑆ࠍ⏕଻ߔࠇ߫↢ሽน⢻
ߣߥࠆ⒟ᐲߢ޽ࠆ(࿑ 3)34)‫ޕ‬୘ੱ⊛ߦߪ‫⷗ߩߎޔ‬Ⓧ߽
ࠅߪ޽߹ࠅߦਇ⏕ቯⷐ⚛ߩᄢ߈ߥ઒ቯ߇ዉ౉ߐࠇߡ
߅ࠅ‫৻ޔ‬ంᐕߩ࠲࡯ࡦࠝ࡯ࡃ࡯ㅦᐲߣ޿߁ౕ૕⊛ߥᢙ୯ߪ޽߹ࠅାᙀᕈ߇ߥ޿ߣ⠨߃ࠆ‫ߒ߆ߒޕ‬㊀ⷐߥߩߪ‫ޔ‬Ⴧᱺߣ↢ሽߦ߅޿ߡᔅⷐߥઍ⻢ㅦᐲ߇ᭂ߼ߡ
Ꮕ߇ᄢ߈޿ߣ޿߁ߎߣߢ޽ࠅ‫ޔ‬ૐ᷷ⅣႺߢ޽ࠇ߫↢๮ᵴേߩ⛽ᜬߪᭂ߼ߡኈᤃߢ޽ࠆߣ޿߁ߎߣࠍ᣿⏕ߦ␜ߒߚὐߢ޽ࠈ߁‫ޕ‬ ৻ᣇ McCollom and Amend19)ߪ‫⚦ޔ‬⢩ࠍ᭴ᚑߔࠆ᦭
ᯏ‛ࠍߔߴߡวᚑ(ⶄ⵾)ߔࠆߩߦᔅⷐߥᦨዊ㒢ߩࠛ
ࡀ࡞ࠡ࡯㊂ࠍ‫⚦ޔ‬⢩ࠍ᭴ᚑߔࠆ᦭ᯏ‛ߩ⚵ᚑߣሽ࿷㊂߅ࠃ߮⃻࿷೑↪น⢻ߥ᦭ᯏ‛วᚑൻቇ෻ᔕߩᾲജ
ቇ⊛࠺࡯࠲ࡌ࡯ࠬ߆ࠄㄭૃ⸘▚ߦࠃߞߡ⷗Ⓧ߽ߞߚ‫ޕ‬ᓐࠄߩ⸘▚ߦࠃࠆߣ‫ޔ‬1g ߩ⚦⢩ࠍቢోߦⶄ⵾ߔࠆઍ
⻢ߦᔅⷐߥࠛࡀ࡞ࠡ࡯㊂ߪ‫ޔ‬ᅢ᳇᧦ઙਅߢߪ 18.4 KJ‫ޔ‬ህ᳇᧦ઙਅߢߪ 1.4 KJ ߢ޽ࠆߣ⷗Ⓧ߽ࠄࠇߚ‫ߩߎޕ‬⎇ⓥߩᭂ߼ߡ㊀ⷐߥὐߪ‫↢ޔ‬๮ᵴേߩ⛽ᜬߦᔅⷐߥ
ᦨዊߩࠛࡀ࡞ࠡ࡯㊂ࠍ‫ޔ‬ᬌ಴ߦ⛘ኻ⊛ߥ㒢⇇ߩ޽ࠆ᷹ⷰ࡮ಽᨆ࡮ቯ㊂ߦଐሽߔࠆࡄ࡜ࡔ࡯࠲࡯(଀߃߫‫ޔ‬ઍ⻢ᵴᕈㅦᐲ߿⚦⢩ኒᐲ߿ൻቇ‛⾰ߩỚᐲ)ࠍ૶ࠊ
ߕߦ‫ޔ‬ൻቇ෻ᔕߦ࿕᦭ߩᾲജቇ⊛ߥࡄ࡜ࡔ࡯࠲࡯ࠍ
૶߁ߎߣߦࠃߞߡ‫ޔ‬ೋ߼ߡቯ㊂⊛ߥ ME ߩ✚㊂ࠍ⷗
Ⓧࠆߎߣ߇ߢ߈ߚὐߢ޽ࠆ‫↢ޕ‬๮ᵴേߩ⛽ᜬߦᔅⷐߥᦨዊߩࠛࡀ࡞ࠡ࡯✚㊂ߩౕ૕⊛ߥ୯߇␜ߐࠇߚߎ
ߣߦࠃߞߡ‫↢ޔ‬๮ߩᦨዊන૏ߢ޽ࠆ⚦⢩ߩ㊂ߣߘߩࠛࡀ࡞ࠡ࡯㊂߇ଏ⛎ߐࠇࠆߴ߈ᤨ㑆න૏߇઒ቯߐࠇ
ࠇ߫‫ߩߘޔ‬ⅣႺߦ߅ߌࠆ ME ࠍ▚಴ߔࠆߎߣ߇ߢ߈
ࠆࠃ߁ߦߥߞߚ‫ޕ‬ ߎࠇࠄ↢‛ߩࡃࠗࠝࡑࠬߩ⛽ᜬߦᔅⷐߥ⚦⢩ߩઍ⻢࿁ォㅦᐲߣ ME ߩ⷗Ⓧ߽ࠅ߆ࠄ‫ޔ‬࿾⃿ߩ↢‛(ᓸ↢‛⚦⢩)ߩ↢๮ሽ࿷น⢻᧦ઙࠍౕ૕⊛ߦ▚಴ߒߡߺࠆ‫⪺ߡߒߣ଀ޕ‬⠪߇ಽ㔌ߒߚᅢᾲᕈࡔ࠲ࡦ⩶Methanotorris formicicum39)ࠍ⠨߃ࠆ‫⋥ߪ⩶ᧄޕ‬ᓘ 1
μm ߩ⃿⩶ߢ‫ᦨޔ‬ㆡ᧦ઙߢߪ 70 C ߢ 30 ಽߩ୚ടᤨ
㑆ࠍ᦭ߔࠆ᳓⚛⾗ൻࡔ࠲ࡦ⩶ߢ޽ࠆ‫ ⩶ᧄޕ‬1 ⚦⢩޽ߚࠅߩᐔဋ⾰㊂ߪ 4.7 x 10-12 g ߢ޽ࠅ‫⩶ࡦ࠲ࡔߩߎޔ‬ߩ 1 ⚦⢩߇‫ ߫߃଀ޔ‬25 C ߢ⥄Ꮖ⛽ᜬߔࠆߚ߼ߦᔅ
ⷐߥࠛࡀ࡞ࠡ࡯㊂߇‫ޔ‬McCollom and Amend19)ߩ⷗Ⓧ߽ࠅߢ޽ࠆ 1.4 KJ/gcells ߩ୯߆ࠄ 6.6 x 10-9 J ߣ▚಴ߢ߈ࠆ‫ޕ‬ᰴߦ Price and Sower29)ߩ 25C ߦ߅ߌࠆࠕࠬ
ࡄ࡜ࠡࡦ㉄ߩ࡜࠮ࡒൻㅦᐲߦ඘ᢜߔࠆઍ⻢ㅦᐲߪ 3
x 10-6 gC/gC•h ߣ઒ቯߢ߈ࠆ‫ ࠅ߹ߟޕ‬3 x 106 ᤨ㑆ߢ
6.6 x 10-9 J ߩࠛࡀ࡞ࠡ࡯ࠍᓧࠄࠇࠇ߫‫ࡦ࠲ࡔߩߎޔ‬⩶ 1 ⚦⢩ߪ 25 C ߢ↢๮ᵴേࠍ⛽ᜬߢ߈ࠆߣ޿߁⸘
▚߇ᚑࠅ┙ߟ‫ߪ⩶ࡦ࠲ࡔߩߎޕ‬᳓⚛ߣੑ㉄ൻ὇⚛߆ࠄࠛࡀ࡞ࠡ࡯ࠍ₪ᓧߒ‫߫߃଀ޔ‬ᮡḰ⁁ᘒߢ 4 mol ߩ᳓⚛ߣ 1 mol ߩੑ㉄ൻ὇⚛߆ࠄᓧࠄࠇࠆࠛࡀ࡞ࠡ࡯
ߪ 1.3 x 105 J ߢ޽ࠆ‫⩶ࡦ࠲ࡔߩߎߡߞࠃޕ‬㧝⚦⢩߇
25C ߢ↢๮ࠍ⛽ᜬߔࠆߚ߼ߦߪ‫ޔ‬㧝⚦⢩ⓨ㑆ߦ 340ᐕ㑆ߦ᳓⚛߇ 2 x 10-13 mol‫ޔ‬ੑ㉄ൻ὇⚛߇ 0.5 x 10-13
mol ⒟ᐲଏ⛎ߐࠇࠆᔅⷐ߇޽ࠆߣ⸒߁ߎߣߢ޽ࠆ‫ޕ‬߽ߜࠈࠎߎߩ႐ว‫ޔ‬ᵹ૕߇േߊߎߣߦࠃߞߡ߽‫ߘޔ‬ߩ႐ߢߩൻቇ෻ᔕߢ↢ᚑߐࠇߡ߽ࠃ޿‫ߩߘߡߒߘޕ‬Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ቮⅣႺߦ߅ߌࠆᦨ߽᥉ㆉ⊛ߥࠛࡀ࡞ࠡ࡯Ḯߣߒߡߩ᳓⚛ଏ⛎㊂߆ࠄ⠨߃ߚౕ૕⊛ߥ↢๮ሽ࿷น⢻᧦ઙߩ
⷗Ⓧ߽ࠅߪ‫ޔ‬࿾⃿ߦ߅޿ߡࠛࡀ࡞ࠡ࡯⺰⊛ߦ↢๮߇↢ሽ㧔ዋߥߊߣ߽↢๮ᵴേࠍ⛽ᜬ㧕ߢ߈ߥ޿ⅣႺࠍ
⷗ߟߌࠆ੐߇㔍ߒ޿ߎߣࠍ᣿⏕ߦ␜ߔ߽ߩߢ޽ࠆ‫ޕ‬᳓⚛ଏ⛎㊂ߪ‫ޔ‬࿾⃿⴫ጀߩ(ᶏᵗ߿࿾Ზ਄ㇱࠍ฽߻)
᳓࿤ⅣႺߦ߅޿ߡ‫ޔ‬ਥⷐߥ㕖↢‛ቇ⊛᳓⚛↢ᚑࡔࠞ࠾࠭ࡓ‫⚉Ⱜ߫߃଀ޔ‬ጤൻ෻ᔕ 2)‫ޔ‬᳓ߩ᡼኿✢ಽ⸃ 4)‫ޔ‬ጤ⍹⎕უ߿៺ᡂߦ઻߁ᯏ᪾ൻቇ෻ᔕ 9)‫ߞࠃߦߤߥޔ‬ߡኈᤃߦ㆐ᚑߐࠇࠆߣ⠨߃ࠄࠇࠆ‫ࠎࠈߜ߽ޕ‬᳓⚛ߛߌ߇↢๮ᵴേߩࠛࡀ࡞ࠡ࡯Ḯߢߪߥ޿‫ޔߒ߆ߒޕ‬ቝ

࿑ 3  Price and Sower29)ߦࠃࠆ᷷ᐲߣᓸ↢‛ߩჇᱺ(growth)‫⛽ޔ‬ᜬ(maintenance)‫↢ޔ‬ሽ(survival)ߦᔅⷐߥઍ⻢ㅦᐲ(਎ઍ੤ઍㅦᐲ)ߩ㑐ଥߦߟ޿ߡߩᮨᑼ࿑(ේ࿑ࠍᡷᄌ)‫↢ޕ‬ሽ(survival)ߦᔅⷐߥઍ⻢ㅦᐲߪ‫ޔ‬ዋߥߊߣ߽ Lࠕࠬࡄ࡜ࠡࡦ㉄ߩ⥄ὼ࡜࠮ࡒൻㅦᐲࠍ਄࿁ࠆᔅⷐ߇޽ࠆ‫ޕ‬
 McCollom and Amend19)ߦࠃߞߡ⷗Ⓧ߽ࠄࠇߚ MEߪ‫ޔ‬ታ㓙ߩᓸ↢‛ߩჇᱺታ㛎߆ࠄ⷗Ⓧ߽ࠄࠇࠆ ME
ߦᲧߴߡ㆔߆ߦዋߥ޿ࠛࡀ࡞ࠡ࡯㊂ߢ޽ࠆߎߣ߇⍮ࠄࠇߡ޿ࠆ‫ޔ߫߃଀ޕ‬਄⸥ߩᅢᾲᕈࡔ࠲ࡦ⩶Methanotorris formicicum ߩ 1 ⚦⢩߇⥄Ꮖ⛽ᜬߔࠆߚ߼ߦᔅⷐߥࠛࡀ࡞ࠡ࡯㊂ߪ‫ޔ‬McCollom and Amend19)
ߩ ME ࠍ↪޿ߚ႐ว‫ޔ‬6.6 x 10-9 J ߢ޽ߞߚ‫৻ޕ‬ᣇ‫ޔ‬Takai et al.39)ߩၭ㙃ታ㛎ߦ߅޿ߡ‫ޔ‬1 ⚦⢩߇୚ടߔࠆ
ߩߦᔅⷐߥ᳓⚛ᶖ⾌㊂߆ࠄ⷗Ⓧ߽ࠄࠇࠆࠛࡀ࡞ࠡ࡯
㊂ߪ‫ ߘࠃ߅ޔ‬4.9 x 10-3 J ߢ޽ࠆ‫ࠎࠈߜ߽ޕ‬ᭂ߼ߡࠛࡀ࡞ࠡ࡯⺰⊛ߦ᦭೑ߥၭ㙃᧦ઙਅߢᓸ↢‛ߪࠛࡀ࡞ࠡ࡯ઍ⻢ߦࠃߞߡᓧࠄࠇߚࠛࡀ࡞ࠡ࡯ࠍߔߴߡჇᱺ
ߦ↪޿ࠆࠊߌߢߥߊ‫࡯ࠡ࡞ࡀࠛޔ‬ᶖ⾌߇㕖ല₸⊛ߢ޽ࠆߎߣߪ੍ᗐ߇ߟߊ‫߽ߘ߽ߘޔߦࠄߐޕ‬Methanotorris formicicum ߩࠃ߁ߥᅢᾲᕈࡔ࠲ࡦ⩶ߪ‫ޔ‬102ࠛࡀ࡞ࠡ࡯Ḯߢ޽ࠆ᳓⚛߇ᭂ߼ߡ⼾ንߥᷓᶏᾲ᳓ⅣႺߦ↢ᕷߔࠆᓸ↢‛ߢ޽ࠅ‫↢ߊࠄߘ߅ޔ‬⢒ߩߚ߼ߩࠛࡀ࡞ࠡ࡯ല₸߇ᦨ߽ᖡ޿ᓸ↢‛ߩࠣ࡞࡯ࡊߢ޽ࠆ
ߣ⠨߃ࠄࠇࠆ‫ޔߕࠄࠊ߆߆߽ߦޕ‬ᾲജቇ⊛⸘▚ߦࠃ
ߞߡ⷗Ⓧ߽ࠄࠇߚ ME ߣታ㓙ߩታ㛎ߦ߅޿ߡⷰኤߐ
ࠇࠆ↢⢒ߦᔅⷐߥࠛࡀ࡞ࠡ࡯㊂ߩᩰᏅߪᭂ߼ߡᄢ߈
޿‫੹ޕ‬ᓟ‫↢ߩ‛↢ޔ‬⢒߿↢ሽߦߣߞߡᭂ߼ߡ෩ߒ޿
ᭂ㒢ⅣႺߦ߅ߌࠆࠛࡀ࡞ࠡ࡯⺰߆ࠄ⷗ߚ↢๮ሽ࿷᧦ઙߩ⎇ⓥ߇ㅴ߻ߎߣߦࠃߞߡ‫ߩߘޔ‬ᩰᏅࠍߤߎ߹ߢ
❗ዊߔࠆߎߣ߇ߢ߈ࠆ߆ࠍ⷗ᭂ߼ࠆᔅⷐ߇޽ࠆ‫޿ޕ‬ߕࠇߦߖࠃ‫ߦ⺰࡯ࠡ࡞ࡀࠛޔ‬ၮߠ޿ߚ↢๮ሽ࿷น⢻ᕈߩℂ⸃߇‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮߿↢๮࿤ߩ㒢⇇߿࿾⃿ᄖ↢๮ߩሽ࿷ߩតᩏߦ߅޿ߡ‫ޔ‬ᭂ߼ߡ㊀ⷐߥ᭎ᔨ߆ߟ⎇ⓥᚢ⇛ߣߥࠆߎߣߪ㑆㆑޿ߥ޿‫ޕ‬

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)੹ᓟߩዷᦸ ᧄ⺰ߢߪ‫ޔ‬࿾⃿↢๮ߩ㒢⇇߿↢๮࿤ߩ㒢⇇ࠍ᣿ࠄ߆ߦߔࠆ⎇ⓥߩ೔㆐ὐ߿໧㗴ὐߦߟ޿ߡ⚫੺ߒߡ߈ߚ‫ޕ‬࿾⃿↢๮ߩ㒢⇇߿↢๮࿤ߩ㒢⇇ࠍ᣿ࠄ߆ߦߔࠆ
⎇ⓥߪ‫ޔ‬዁᧪ߩ࿾⃿ᄖᖺᤊ߿ⴡᤊߦ߅ߌࠆ↢๮តᩏߦਇนᰳߩ⑼ቇℂ⺰߿⎇ⓥᚢ⇛‫ޔ‬ಽᨆᛛⴚ߿ᣇᴺ⺰ߩᩮᐙࠍ᭴▽ߔࠆߛߌߢߥߊ‫ޔ‬‫↢ޟ‬๮ߣߪ૗߆‫ࠍޠ‬ℂ⸃ߔࠆၮ⋚ࠍᚑߔ߽ߩߢ޽ࠆ‫ޕ‬ ߒ߆ߒ࿾⃿↢๮ߩ㒢⇇߿↢๮࿤ߩ㒢⇇ࠍ᣿ࠄ߆ߦ
ߔࠆ⎇ⓥߦߪ߹ߛ߹ߛస᦯ߔߴ߈໧㗴߇ሽ࿷ߔࠆ‫ޕ‬଀߃߫ᚒ‫ߩߎޔߪޘ‬࿾⃿ߦ߅޿ߡᧂߛ↢๮ሽ࿷ⅣႺߣ↢๮㕖ሽ࿷ⅣႺߩႺ⇇߿ߘߩ᧦ઙߦߟ޿ߡ‫ౕޔ‬૕⊛ߥ⸽᜚ࠍ᣿␜ߔࠆߎߣ߇ߢ߈ߡ޿ߥ޿‫ޔߚ߹ޕ‬ᶏᐩਅ࿾ᲖౝⅣႺߦߪ࿾⃿ߦ߅ߌࠆᦨᄢߩẜ࿷ࡃࠗࠝࡑࠬ߇ሽ࿷ߔࠆߣ޿߁઒⺑ߩឭ␜ 41)߆ࠄ 10 ᐕએ਄⚻ㆊߒߡ‫⷗ߩߘޔ‬Ⓧ߽ࠅ߇ᄢ߈ߊਅᣇୃᱜߐࠇ 15)‫ޔ‬ᧂߛߘߩᱜ⏕ߥࡃࠗࠝࡑࠬࠍቯ㊂ߔࠆߦ⥋ߞߡ޿ߥ޿‫ޕ‬ห᭽ߦ‫ޔ‬࿾ᲖౝⅣႺߦߪ࿾⃿⴫ጀⅣႺߣߪ♽⛔
⊛ߦᄢ߈ߊ⇣ߥࠆᄙ᭽ߥᓸ↢‛߇ሽ࿷ߒߡ޿ࠆߎߣ
ࠍ␜ߒߚߦ߽߆߆ࠊࠄߕ 13)‫ߩߘޔ‬ఝභᓸ↢‛⟲ߩታ㓙ߩᯏ⢻ࠍ⃻႐ⅣႺߦ߅޿ߡᝒ߃ࠆߎߣ߇ߢ߈ߕ‫ޔ‬ߘࠇߤߎࠈ߆ᯏ⢻ߩᄙߊߪᧂ⸃᣿ߩ߹߹ߢ޽ࠆ‫ޕ‬ ‫↢ޟ‬๮ߣߪ૗߆‫ࠍޠ‬⍮ࠆߚ߼ߦߪ‫ޔ‬ᚒ‫߇ޘ‬໑৻⍮ߞߡ޿ࠆ‫ޟ‬࿾⃿↢๮‫ߊࠄߘ߅ޔߊߥߢߌߛޠ‬ቝቮߦ߅޿ߡ߽ᐞᄙߦሽ࿷ߔࠆߣ⠨߃ࠄࠇࠆ࿾⃿ᄖ↢๮ߩ޽ࠅᣇߣߩᲧセࠍㅢߓߡ‫ޔ‬ቝቮ౒ㅢේℂߣߒߡߩ‫↢ޟ‬๮⺰‫ࠍޠ‬᭴▽ߔࠆᔅⷐ߇޽ࠆߣ޿߁ࠕࠬ࠻ࡠࡃࠗࠝ
ࡠࠫ࡯ߩᄢ߈ߥℂᔨߪᭂ߼ߡ⺑ᓧജߦንߺ‫ߩߘޔ‬ℂ
ᔨ߿⑼ቇ⋡ᮡߪ᳞ᔃജߩ޽ࠆ㝯ജ⊛ߥ⎇ⓥ࠹࡯ࡑ߿
⎇ⓥࡊࡠࠫࠚࠢ࠻ࠍ↢ߺ಴ߔේേജߢ޽ࠆ‫ߒ߆ߒޕ‬৻ᣇߢ‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮ࠍචಽߦℂ⸃ߔࠆߎߣߥ
ߊ‫ޔ‬࿾⃿ᄖ↢๮ࠍតᩏ࡮⼂೎࡮ᬌ಴ߔࠆߎߣߪ৻⒳ߩ⾲ߌߢߒ߆ߥߊ‫ߟ߆ޔ‬ᄢ߈ߥℂᔨࠍ㆐ᚑߔࠆߎߣ
ߪਇน⢻ߢ޽ࠈ߁‫⪺ޕ‬⠪ߪ‫ޔ‬࿾⃿ߦ߅ߌࠆ↢๮࿤ߣ㕖↢๮࿤ߩႺ⇇ߦ↢ሽߔࠆᭂ㒢ⅣႺ↢‛ߦኻߔࠆᧄ
⾰⊛ߥℂ⸃ࠍㅊ᳞ߔࠆߎߣߎߘ‫ߩߘޔ‬ᄢ߈ߥℂᔨ߿⑼ቇ⋡ᮡߦ೔㆐ߔࠆߚ߼ߩᦨ߽㊀ⷐߥ╙৻ᱠߢ޽ࠆ
ߣାߓߡ޿ࠆ‫ޕ‬

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83: 9542-9546.

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ᭂ㒢ⅣႺ↢‛ቇળ╙ 13 ࿁ᐕળႎ๔
╙ 13 ࿁ᐕળߪ‫ޔ‬12 ᦬ 1-2 ᣣ㧔࿯࡮ᣣ㧕ߦᣣᧄᄢቇᢥℂቇㇱ㧔਎↰⼱඙᪉਄᳓㧕‫ޔ‬3 ภ㙚 5 㓏ߦߡ㐿௅ߐࠇ߹ߒߚ‫ޕ‬ᒰᣣߪ‫ޔ‬ᄙߊߩળຬ࡮㕖ળຬߩ⊝᭽ߦߏෳ㓸޿ߚߛ߈‫ޔ‬ญ㗡⊒⴫ 23 ઙ‫ ⴫⊒࡯࠲ࠬࡐޔ‬69 ઙߩ৻⥸⊒⴫ߩઁ‫ޔ‬᳇㍈ߩᣣᄢߩ 3 ฬߩవ↢ᣇߦࠃࠆ․೎࠮࠶࡚ࠪࡦ㧔ᣣᄢ࠮࠶࡚ࠪࡦ㧕ࠍᐕળࠝ࡯ࠟ࠽ࠗࠩ࡯ߩὑᚒ
੗వ↢㧔ᣣᄢ㧕ߦડ↹ߒߡ޿ߚߛ߈‫᧲ޔߦᦝޔ‬੩⮎⑼ᄢߩᷰㆺవ↢ߦࠃࠆ․೎⻠Ṷ‫ޔ‬ቇળᅑബ⾨ฃ⾨⻠Ṷ 2ઙߣᄢᄌ⋓ࠅߛߊߐࠎߥળߣߥࠅ߹ߒߚ‫ޕ‬ෳട⠪ว⸘ߪ 273 ฬ㧔ౝ⸶ࠍએਅߩ⴫ߦ␜ߔ㧕ߣߥࠅ‫ޔ‬೨࿁ߩ㐳ፒᐕળෳട⠪ 204 ฬࠍᄢ߈ߊ਄࿁ࠆෳട⠪ੱᢙߣߥࠅ߹ߒߚ‫ޕ‬ᐕળߩ߅਎⹤ࠍ޿ߚߛ߈߹ߒߚ‫ޔ‬ᣣᧄᄢቇ࡮
ὑᚒ੗వ↢㧔ቇⴚᐙ੐㧕ߦߪ‫ޔ‬ᐕળ㐿௅ߦะߌ♖ജ⊛ߦᅁ⿛ߒߡ޿ߚߛ߈߹ߒߚ‫ޔߢߎߎޕ‬ὑᚒ੗వ↢ߦᗵ⻢↳ߒ਄ߍࠆߣߣ߽ߦ‫ޔ‬ᒰᣣߏෳട޿ߚߛ޿ߚ⊝᭽ᣇߦ੐ോዪ৻หᔃࠃࠅᗵ⻢↳ߒ਄ߍ߹ߔ‫ޕ‬

৻⥸
ቇ↢
✚⸘

੐೨↳ߒㄟߺੱᢙ
83
㧔ળຬ 76‫ޔ‬㕖ળຬ 7㧕
131
㧔ળຬ 23‫ޔ‬㕖ળຬ 108㧕214㧔ળຬ 99‫ޔ‬㕖ળຬ 115㧕ᒰᣣ↳ߒㄟߺੱᢙ
36
㧔ળຬ 9‫ޔ‬㕖ળຬ 27㧕
23
㧔ળຬ 0‫ޔ‬㕖ળຬ 23㧕
59
㧔ળຬ 9‫ޔ‬㕖ળຬ 50㧕ว⸘119
㧔ળຬ 85‫ޔ‬㕖ળຬ 34㧕
154
㧔ળຬ 23‫ޔ‬㕖ળຬ 131㧕273
㧔ળຬ 108‫ޔ‬㕖ળຬ 165㧕 ੹࿁߽‫ޔ‬ડᬺᛛⴚࠦ࡯࠽࡯ߦߪએਅߩ 6 ␠ߦߏෳട޿ߚߛ߈߹ߒߚ‫ޕ‬๺శ⚐⮎Ꮏᬺᩣᑼળ␠
ᣣᧄ㔚ሶᩣᑼળ␠
ኅ↰⾏ᤃᩣᑼળ␠+╳ᵄኅ↰ᩣᑼળ␠࡜ࠗࠞ ࡑࠗࠢࡠࠪࠬ࠹ࡓ࠭ᩣᑼળ␠
ᩣᑼળ␠࠻ࡒ࡯♖Ꮏᣣ㋕૑㊄ⅣႺᩣᑼળ␠ ቇળ੐ോዪࠃࠅ‫ޔ‬ᐕળ߳ߩߏදജߦ㑐ߒߡᔃࠃࠅᗵ⻢↳ߒ਄ߍ߹ߔ‫ޕ‬‫․ޣ‬೎⻠Ṷ‫ޤ‬᧲੩⮎⑼ᄢቇߩᷰㆺ৻຦వ↢ߦ‫ߩ↢౒ޟ‬ᭂߺ‫ߣޠ‬㗴ߒߡ‫⻠ߏޔ‬Ṷࠍ޿ߚߛ߈߹ߒߚ‫ޕ‬ᓸ↢‛㔚ᳰߦ㑐ߔࠆߏ⥄りߩ⎇ⓥ߆ࠄ‫ޔ‬ᓸ↢‛㑆ߩ⋧੕૞↪‫⥝ߏߦ⽎⃻ߚߞ޿ߣ↢౒ޔ‬๧ࠍ߽ߚࠇ‫ޔ‬ᓸ↢‛ߩ⢻ജࠍ೑↪ߒߚ㔚᳇౒↢ߥߤᰴ਎ઍࠛ࠽ࠫ࡯߳ߣะ߆߁␠ળߩⷐ⺧ߦขࠅ⚵߻ᦨవ┵ߩߏ⎇ⓥࠍߏ⚫੺޿ߚߛ߈߹ߒߚ‫ޕ‬ὑᚒ੗వ↢‫⮮ޔ‬ේవ↢‫ޔ‬ᷰㆺవ↢105

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)‫ޣ‬ᣣᄢ࠮࠶࡚ࠪࡦ‫ޤ‬ὑᚒ੗వ↢ߩડ↹ߦࠃࠅ‫ޔ‬એਅߩ 3 ฬߩ᳇㍈ߩవ↢ᣇߦࠃࠆߏ⻠Ṷࠍ޿ߚߛ߈߹ߒߚ‫ޕ‬Ỉ↰ඳมవ↢㧔ᣣᧄᄢቇᢥℂቇㇱ✚วᢥൻ⎇ⓥቶ㧕‫ߩࠦࠗࠞޟ‬ભ⌁ߣભ⌁㒖ኂߩಽሶᯏ᭴‫ޠ‬ᯅᧄિ຦వ↢㧔ᣣᧄᄢቇᢥℂቇㇱൻቇ⑼㧕
‫ޟ‬ᓸ↢‛ߦࠃࠆࡂࡠࠞ࡯ࡏࡦߩ↢ᚑ‫ޠ‬⩲ේᱜ㓶వ↢㧔ᣣᧄᄢቇᢥℂቇㇱൻቇ⑼࡮2012 ᐕᐲᣣᧄಽᨆൻቇળቇળ⾨㧕‫ޟ‬ᓸዊ㉂⚛࠮ࡦࠨ࡯ߦࠃࠆູ੃㘃ᄢ⣖ᶏ㚍ਛߩࠣ࡞࠲ࡒࡦ㉄ߩ⸘᷹‫ޠ‬‫ޣ‬ᅑബ⾨ฃ⾨⻠Ṷ‫ޤ‬
2012 ᐕᐲߩቇળᅑബ⾨ߪᰴߩ 2 ฬߦ᳿ቯߒ‫ޔ‬ฃ⾨⻠Ṷࠍ޿ߚߛ߈߹ߒߚ‫ޕ‬࡮੗਄⻞๋ వ↢㧔ችፒᄢቇ࡮IR ផㅴᯏ᭴㧕‫ޟ‬ᓸ↢‛ߩ㔚ሶવ㆐ߦࠃࠆⅣႺᳪᨴ‛⾰ಽ⸃ߣ⊒㔚‫ޠ‬࡮ㇹ↰⑲৻㇢వ↢㧔㐳ፒᄢ㒮࡮Ꮏቇ⎇ⓥ⑼㧕‫⿥ޟ‬ᅢᾲ⩶↱᧪ࠣ࡞࠲ࡒࡦ㉄⣕᳓⚛㉂⚛ߩᾲᵴᕈൻᯏ᭴ߩ⸃᣿‫ޠ‬੗਄ஜ๋వ↢

ㇹ↰⑲৻㇢వ↢੗਄వ↢ߪ‫ޔ‬ᄢቇ㒮↢ᤨઍᒰቇળߢ૗࿁߽ߏ⊒⴫ߐࠇߡ߅ࠅ‫ߩߘޔ‬ᓟ☨࿖⇐ቇࠍ⚻ߡᄢ߈ߊᚑ㐳ߐࠇ‫ޔ‬ᒰቇળߦᏫߞߡ߈ߡ޿ߚߛ޿ߚవ↢ߢߔ‫ߚ߹ޕ‬ㇹ↰వ↢ߪ‫ޔ‬2002 ᐕᐲߦⴕࠊࠇߚᓼፉᐕળߢ㐿௅ࠍࠨࡐ࡯࠻ߒߡ
޿ߚߛ޿ߡએ᧪‫ޔ‬ᒰቇળߢߏᵴべ޿ߚߛ޿ߡ߅ࠅ߹ߔ‫੹ޕ‬࿁ߩߏฃ⾨ᄢᄌߦ߅߼ߢߣ߁ߏߑ޿߹ߒߚ‫ޕ‬‫ޣ‬ఝ⑲ࡐࠬ࠲࡯⾨‫ޤ‬ఝ⑲ࡐࠬ࠲࡯⾨ߪ‫ޔ‬ክᩏຬߦࠃࠆ෩ᱜߥክᩏߩ⚿ᨐ‫ޔ‬એਅߩ 6 ฬߦ᝼⾨ߐࠇ߹ߒߚ‫ޕ‬
P-20 ᰏ↰ථᔒ㧔᧲ᵗᄢቇᄢቇ㒮࡮↢๮⑼ቇ⎇ⓥ⑼㧕Thermococcus kodakarensis ߩ polB ㆮવሶ⎕უᩣߩ⴫⃻ဳ
⸃ᨆ
P-32 ựፉ⵨ノ㧔┙ᢎᄢቇᄢቇ㒮㨯↢๮ℂቇ㧕ᷓᶏ↱᧪⛘ኻᅢ࿶⩶↱᧪ߩࠗ࠰ࡊࡠࡇ࡞࡝ࡦࠧ㉄⣕᳓⚛㉂⚛ߩ⠴࿶ᕈߪ 1 ࠕࡒࡁ㉄ߦ⿠࿃ߔࠆ
P-42 ౷↰ብ⍫㧔੩ㇺᄢቇᄢቇ㒮 Ꮏቇ⎇ⓥ⑼㧕ࠕ࡯ࠠࠕߩ⵬㉂⚛ A ↢วᚑߦ㑐ߔࠆ㉂⚛ቇ⊛⸃ᨆ߅ࠃ߮೙ᓮ
ᯏ᭴ߩ⸃᣿
P-46 ዊ㊁ኹᆭᢥ㧔᧲੩⮎⑼ᄢ㒮࡮↢๮⑼ቇ㧕Deinococcus ߣ Thermus ߦ౒ㅢߔࠆᣂⷙ DNA ㎮㑆᨞ᯅୃᓳㆮવ
ሶߩㆮવቇ⊛⸃ᨆ
P-58 ᧻Ỉᷕ㧔᧲੩ᄢቇ࡮↢‛↢↥Ꮏቇ⎇ⓥ࠮ࡦ࠲࡯㧕Carbazole 1,9a-dioxygenase ߦ߅ߌࠆ㉄ൻ㉂⚛-ferredoxin㑆ߩ⚿ว⸃㔌ࡔࠞ࠾࠭ࡓߩ⸃᣿
P-64 ⧯੗ᥙ㧔ᐢፉᄢቇᄢቇ㒮࡮↢‛࿤⑼ቇ⎇ⓥ⑼㧕㜞ᐲᅢႮᕈ⩶ Haloarcula japonica ߩࡇࡠ࡝ࡦ㉄ട᳓ಽ⸃ߩᾲജቇ⊛ⷞὐ߆ࠄ⷗ߚႮㆡᔕ

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)‫ޣ‬ฬ⹷ળຬ೙ᐲߩഃ⸳‫ޤ‬ᐕᰴ✚ળߦ߅޿ߡ‫ޔ‬ᒰቇળߩฬ⹷ળຬ೙ᐲ*߇ᣂߚߦഃ⸳ߐࠇࠆߎߣ߇ᛚ⹺ߐࠇ߹ߒߚ‫੹ޕ‬ᓟቇળ੐ോዪࠍਛ
ᔃߦㆬ⠨૞ᬺ߇ㅴ߼ࠄࠇ߹ߔ‫ޕ‬*ળೣ╙ 4 ᧦ 3 㗄㧦ฬ⹷ળຬߪ‫ޔ‬ᒰቇળߩ⊒ዷ߿ಽ㊁ߦഞഭߩ޽ߞߚ⠪ߢ‫ޔ‬ᐙ੐ળߦࠃࠆផ⮈‫⼏⹏ޔ‬ຬળߦࠃࠆᛚ⹺ࠍ⚻ߡ‫ޔ‬ળ㐳ߩᜰฬߦࠃࠅ᳿ቯߐࠇ‫ޔ‬ᒰቇળߩ௅ߔࠪࡦࡐࠫ࠙ࡓ࡮ᐕળ╬ߦ᜗ᓙߐࠇࠆ‫ޕ‬‫ޣ‬2013 ᐕᐲࠃࠅቇળ੐ോዪ߇ᄌᦝߣߥࠅ߹ߔ‫ޤ‬ߎࠇ߹ߢߩ੗਄ᐙ੐㐳㧔᧲ᵗᄢ㧕࡮ട⮮ᐼോᐙ੐㧔JAMSTEC㧕ߩቇળ੐ോዪ૕೙߇‫ޔ‬2013 ᐕᐲࠃࠅ⧯㄰ࠅ߹
ߔ‫ޕ‬ળ㐳ᜰฬߦࠃࠅ‫ޔ‬ਛ᧛⡡ᐙ੐㐳㧔᧲Ꮏᄢ㧕࡮દ⮮᡽ඳᐼോᐙ੐㧔᧲ᵗᄢ㧕ࠍਛᔃߦᣂቇળ੐ോዪ߇✬ᚑߐࠇ‫ޔ‬੐ോዪࠍ᧲Ꮏᄢߔߕ߆ߌบࠠࡖࡦࡄࠬߦ⒖ⴕߔࠆߎߣ߇㧘✚ળߦߡႎ๔ߐࠇ߹ߒߚ‫ޕ‬ψᣂ੐ോዪߩㅪ⛊వ㧦‫ޥ‬226-8501 ᮮᵿᏒ✛඙㐳ᵤ↰↸ 4259-J2-13᧲੩Ꮏᬺᄢቇ ᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼↢‛ࡊࡠ࠮ࠬኾ᡹ ਛ᧛⡡⎇ⓥቶౝᭂ㒢ⅣႺ↢‛ቇળ੐ോዪ ਛ᧛ ⡡ (ᐙ੐ળᐙ੐㐳)
FAX: 045-924-5837 E-mail: kyokugenjm10@extremophiles.jp
ᭂ㒢ⅣႺ↢‛ቇળ੐ോዪ

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ቇળ⹹ᛩⓂⷙቯ
ᭂ㒢ⅣႺ↢‛ቇળ⹹ᛩⓂⷙቯ㧝㧚ᛩⓂ⠪ߩ⾗ᩰᭂ㒢ⅣႺ↢‛ቇળ⹹㧔ISSN 1348-5474㧕 (Journal ofJapanese Society for Extremophiles)㧔એਅ‫޿ߣ⹹ᧄޔ‬߁㧕߳ߩᛩⓂߪᭂ㒢ⅣႺ↢‛ቇળ㧔એਅ‫ᧄޔ‬ቇળߣ޿߁㧕ળຬߦ㒢ࠆ‫ޔ߅ߥޕ‬ㅪฬߦࠃࠆᛩⓂߩ႐วߦߪ‫ޔ‬ዋߥߊߣ߽⪺⠪ߩ৻ੱ߇ᧄቇળળຬߢߥߌࠇ߫
ߥࠄߥ޿‫ޔߒߛߚޕ‬ᣣᧄ࿖એᄖ߆ࠄߩᛩⓂਗ߮ߦᧄቇળ߇ነⓂࠍଐ㗬ߒߚ႐วߪ଀ᄖߣߔࠆ‫ޕ‬ߌઃߌࠆ‫ߩߎޕ‬႐ว‫ޔ‬ᭂ㒢ⅣႺ↢‛ቇળࡎ࡯ࡓࡍ࡯ࠫߦឝタߐࠇߡ޿ࠆᛩⓂⷐ㗔ߦᓥ߁ߎߣ‫ޕ‬‫ޥ‬113-8657 ᧲੩ㇺᢥ੩඙ᒎ↢ 1-1-1 ᧲੩ᄢቇᄢቇ
㒮ㄘቇ↢๮⑼ቇ⎇ⓥ⑼ᔕ↪↢๮Ꮏቇኾ᡹㧔᧲ᄢ㒮ㄘ↢⑼ᔕ↢Ꮏߣ⋭⇛น㧕ᭂ㒢ⅣႺ↢‛ቇળ⹹✬㓸ᆔຬ㐳  ⍹ ੗ ᱜ ᴦ ( 㔚 ⹤ 㧦 03-5841-8258, FAX 㧦
03-5841-5272) (e-mail: amishii@mail.ecc.u-tokyo.ac.jp)㧞㧚⺰ᢥߩ⒳㘃
ᧄ⹹߳ߩᛩⓂ⺰ᢥߪ‫ޔ‬ᭂ㒢ⅣႺ↢‛㑐ㅪಽ㊁ߩේ⪺⺰ᢥ㧔ㅢᏱ⺰ᢥ‫ޔ‬⍴⺰ᢥ㧕‫ޔ⺑✚ޔ‬ਗ߮ߦ⻉ႎߣߒ‫ޔ‬޿ߕࠇ߽ᧂ⊒⴫ߩ߽ߩߦ㒢ࠆ‫ޕ‬㧔㧝㧕ේ⪺⺰ᢥ (original papers)㧦ᭂ㒢ⅣႺ↢‛ߦ㑐ߔࠆ⺰ᢥߢએਅߩ㧞ߟߩ޿ߕࠇ߆ߦ⹥ᒰߔࠆ⺰ᢥ‫ޕ‬㧝㧕ㅢᏱ⺰ᢥ (regular papers)㧦ᭂ㒢ⅣႺ↢‛ߦ㑐ߔࠆ⺰ᢥߢ‫⥄ࠇߘޔ‬૕⁛┙ߒߚ‫ޔ‬ଔ୯ߩ޽ࠆ⚿⺰޽ࠆ޿ߪ੐ታࠍ฽߻⺰ᢥ‫ޕ‬㧞㧕⍴⺰ᢥ (short communications)㧦ᭂ㒢ⅣႺ↢‛ߦ㑐ߔࠆ⺰ᢥߢ‫ޔ‬ᢿ ⊛ߥ⚿ᨐߢߪ޽ࠆ߇ᣂߒ޿੐ታ
߿ଔ୯ߩ޽ࠆ࠺࡯࠲ࠍ฽߻⍴޿⺰ᢥ‫ޕ‬㧔㧞㧕✚⺑ (reviews)㧦ᭂ㒢ⅣႺ↢‛ߩ޽ࠆ․ቯߩ⎇ⓥಽ㊁ߦߟ޿ߡ‫◲ޔ‬නߥᱧผ⊛⢛᥊ࠍ฽߼‫ᦨޔ‬ㄭߩㅴᱠࠍⷐ⚂ߒ‫ޔ‬น⢻ߢ޽ࠇ߫‫ޔ‬዁᧪ߩ⎇ⓥᣇะᕈࠍ߽ᜰߒ␜ߒߚ⺰ᢥ‫ޕ‬㧔㧟㧕ࡒ࠾࡟ࡆࡘ࡯ (mini-reviews)㧦✚⺑ߩౝ‫ޔ‬Ყセ⊛⍴ߊ➙߼ߚ߽ߩ‫ޕ‬㧔㧠㧕⻉ႎ㧦Ꮞ㗡⸒‫⎇ޔ‬ⓥᦨ೨✢‫⎇ޔ‬ⓥߩ࡞࡯࠷ࠍ
តࠆ‫ޔ‬ቇળ㑐ㅪ⸥੐╬‫ޕ‬㧡㧚⺰ᢥߩክᩏ
ᛩⓂߐࠇߚ⺰ᢥߩౝ‫ޔ‬ේ⪺⺰ᢥ‫ࡘࡆ࡟࠾ࡒޔ⺑✚ޔ‬࡯ߩክᩏߦߪ‫ޔ‬ᭂ㒢ⅣႺ↢‛ቇળ⹹✬㓸ᆔຬߩౝ㧝ฬ߇⽿છ✬㓸ᆔຬߣߒߡ㑐ਈߔࠆ‫ޕ‬ේ⪺⺰ᢥ߅ࠃ߮✚⺑ߩክᩏߪ‫⽿ޔ‬છ✬㓸ᆔຬ߇ㆡቱᜰฬߔࠆⶄᢙߩ
ᩏ⺒⠪ߦࠃࠅߥߐࠇࠆ‫ޕ‬ឝタߩนุߦ㑐ߔࠆᦨ⚳್ቯߪ‫ޔ‬ᩏ⺒⠪߆ࠄߩᩏ⺒⚿ᨐႎ๔ߦၮߠ߈⽿છ✬㓸ᆔຬ߇ⴕ߁‫ߩߘޕ‬ᓟ‫✬ޔ‬㓸ᆔຬ㐳߇⪺⠪ߦክᩏ⚿ᨐࠍㅢ⍮ߔࠆ‫ޔ߅ߥޕ‬ౝኈ࡮૕ⵙߦ໧㗴߇޽ࠆߣ್ᢿ
ߐࠇߚේⓂߦߟ޿ߡߪ‫⪺ޔ‬⠪ߦࠃࠆୃᱜࠍ᳞߼ࠆ‫ޕ‬ୃᱜࠍ᳞߼ࠄࠇߚේⓂߪ‫ޔ‬㧞ࡩ᦬એౝߦ⽿છ✬㓸ᆔຬߦ㄰ㅍߔࠆߎߣߣߒ‫ޔ‬㧞ࡩ᦬ࠍㆊ߉ߡ㄰ㅍߐࠇߥ
߆ߞߚේⓂߪ‫⪺ޔ‬⠪߇ේⓂࠍขࠅਅߍߚ߽ߩߣߒߡ
ಣℂߔࠆ‫ޔ߅ߥޕ‬ᛩⓂߐࠇߚ⺰ᢥߪ㄰ළߒߥ޿‫ޕ‬㧟㧚⺰ᢥߩᦠ߈ᣇ
⺰ᢥߩ↪⺆ߪᣣᧄ⺆߹ߚߪ⧷⺆ߣߒ‫ޔ‬೎ㅜቯ߼ࠆ‫ޟ‬ේⓂၫ╩ⷐ㗔‫ߦޠ‬ᓥ޿◲ẖߦಽ߆ࠅ߿ߔߊ૞ᚑߔࠆߎ
ߣ‫ߪ⺑✚ޕ‬ේⓂ㧟㧜㗁એౝ‫ޔ‬ㅢᏱ⺰ᢥߪේⓂ㧞㧜㗁એౝ‫ߪ࡯ࡘࡆ࡟࠾ࡒޔ‬ේⓂ㧝㧡㗁એౝ‫ޔ‬⍴⺰ᢥߪේ
Ⓜ㧝㧜㗁એౝߣߔࠆ‫⻉ޔ߅ߥޕ‬ႎߩ㗁ᢙߦ㑐ߒߡߪ✬㓸ᆔຬળߢㆡቱ್ᢿߔࠆ‫ޕ‬㧣㧚೎೚ࠅ೎೚ࠅߪ‫ޔ‬pdf ࡈࠔࠗ࡞ߣߒߡ㈩Ꮣߔࠆ‫ޕ‬㧢㧚ᩞᱜᩞᱜߪೋⓂߩߺ⪺⠪ߦㅍઃߔࠆ‫ޕ‬೔⌕ᣣࠃࠅ㧟ᣣએౝߦ‫ޔ‬ේⓂߣߣ߽ߦᜰቯߐࠇߚㅍઃవߦ㄰ㅍߔࠆߎ
ߣ‫ޔ߅ߥޕ‬ේ⪺⺰ᢥߥࠄ߮ߦ✚⺑ߦ㑐ߒߡߪ‫⪺ޔ‬⠪ᩞᱜᤨߩᢥ┨࡮࿑⴫ߩㅊട߿೥㒰ߪ৻ಾ⹺߼ߥ޿‫ޕ‬㧤㧚⪺૞ᮭᧄ⹹ߦឝタߐࠇߚ⺰ᢥߩ⪺૞ᮭߪᧄቇળߦᏫዻߔࠆ‫ޕ‬㧥㧚ߘߩઁᭂ㒢ⅣႺ↢‛ቇળ⹹ᛩⓂⷙቯߥࠄ߮ߦᭂ㒢ⅣႺ↢‛ቇળ⹹⺰ᢥၫ╩ⷐ㗔ߢਇ᣿ߥㇱಽߦ㑐ߒߡߪ‫ޔ‬ᭂ㒢ⅣႺ↢‛ቇળ⹹✬㓸ᆔຬળߦ߅޿ߡㆡቱኻᔕߔࠆ‫ޕ‬ߥ߅‫ޔ‬ᔅⷐ߇⹺߼ࠄࠇࠆ႐วߦߪ‫ޔ‬ᢥᦠൻࠍㅴ߼ࠆ
߽ߩߣߔࠆ‫ޕ‬㧠㧚⺰ᢥߩฃઃ
⺰ᢥේⓂߪ⻉ႎࠍ㒰߈‫ޔ‬ේⓂේᧄߩઁߦࠦࡇ࡯㧟ㇱࠍઃߌ‫ޔ‬ㅍઃ⁁ߣߣ߽ߦ◲ᤃㇷଢᦠ⇐ߣߒߡਅ⸥ተ
ߦㅍઃߔࠆߎߣ‫ࠆࠃߦࡦࠗ࡜ࡦࠝޔ߅ߥޕ‬ᛩⓂ߽ฃ

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Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)⺰ᢥၫ╩ⷐ㗔ᭂ㒢ⅣႺ↢‛ቇળ⹹⺰ᢥၫ╩ⷐ㗔㧝㧚৻⥸⊛੐㗄㧔㧝㧕࠹ࠠࠬ࠻ࡈࠔࠗ࡞ᒻᑼ߹ߚߪ Microsoft Wordᒻᑼߢ଻ሽߢ߈ࠆࡢ࡯࠼ࡊࡠ࠮࠶ࠨ╬ࠍ↪޿ߡේⓂ
ࠍ૞ᚑߔࠆߎߣ‫ޕ‬㧔㧞㧕࠲ࠗ࠻࡞ࡍ࡯ࠫߦߪ‫ޔ‬1)⺰ᢥ㗴ฬ‫ޔ‬2)⪺⠪ฬ‫ޔ‬3)⪺⠪ᚲዻᯏ㑐ฬ㧔ⶄᢙߩ႐วߪ a, b, c࡮࡮ߢ⴫⸥㧕‫ޔ‬4)࡜ࡦ࠾ࡦࠣ࠲ࠗ࠻࡞‫ޔ‬5)ㅪ⛊ኻᔕ⪺⠪ฬ߅ࠃ߮ߘ
ߩㅪ⛊వ૑ᚲ࡮㔚⹤⇟ภ࡮FAX ⇟ภ࡮e-mail ࠕ࠼࡟ࠬ‫ޔ‬6)ࠠ࡯ࡢ࡯࠼㧔㧡⺆એౝ㧕ࠍ᣿␜ߔࠆߎߣ‫ޕ‬ᣣᧄ⺆ߢ૞ᚑߒߚ⺰ᢥߦߪ‫ࠍ⸶⺆⧷ߩࠄࠇߎޔ‬૬⸥ߔࠆߎߣ‫ޕ‬
㧔㧟㧕⺰ᢥශ೚ߦߪ A4 ↪⚕ߩ 㕙ࠍ૶↪ߒ‫ޔ‬㧝ⴕ㧠㧜ሼ㧔⧷⺆ߩ႐วߦߪ㧝ⴕ㧢㧜ሼ㧕‫ޔ‬㧝㗁㧞㧡ⴕߢශ೚ߔࠆߎߣ߇ᦸ߹ߒ޿‫ޔ࠼࡞࡯ࡏޔࠢ࠶࡝࠲ࠗޕ‬޽ࠆ޿ߪࠬࡕ࡯࡞ࠠࡖࡇ࠲࡞ߦߔࠆ⺆ฏ߇޽ࠆ႐ว
ߪ‫ߢࡠࡊ࡯ࡢޔ‬ᄌ឵ߢ߈ߥ޿႐ว‫⿒ޔ‬ሼߢਅ✢‫ޔ‬ᵄਅ✢‫ޔ‬ੑ㊀ਅ✢ࠍ⸥ߔߎߣ‫ޕ‬㧔㧠㧕⇛⺆࡝ࠬ࠻߇޽ࠆ႐วߦߪ‫ᧄޔ‬ᢥ╙㧝㗁ߦ⣉ᵈߣߒߡ⇛⺆ߩ㗄ࠍ⸳ߌ⴫⸥ߔࠆߎߣ‫ޕ‬න૏♽ߣ⋭⇛⸥ภߪ SI න૏ࠍၮᧄߣߔࠆ‫ޕ‬㧔㧡㧕↢‛ߩ๮ฬ෸߮⸥タߪ International Code of
Nomenclature of Prokaryotes, International Code ofBotanical Nomenclature, International Code ofZoological Nomenclature ߦᓥ߁‫ޕ‬ቇฬ߇ೋ಴ߩ႐วߪ
ోㇱࠍ⸥タߔࠆߎߣ‫ޕ‬ේೣߣߒߡ‫ޔ‬㧞࿁⋡એ㒠ߪዻฬߩ㗡ᢥሼࠍ⇛ภߣߔࠆ߇‫޿ߒࠊࠄ⚗ޔ‬႐วߦߪ⋭⇛ߖߕߦ⸥タߔࠆߎߣ‫ޕ‬㧔㧢㧕‛⾰ฬߩ⴫⸥ߪએਅߩⷙቯߦᓥ߁ߎߣ‫ޕ‬㧝㧕᦭ᯏൻว‛ߩ⴫⸥߅ࠃ߮᡼኿ᕈห૏ర⚛ߦࠃࠆᮡ⼂ൻว‛ߩ⴫⸥ߪ IUPAC ⷙቯߦᓥ߁ߎߣ‫ޕ‬㧞㧕↢ൻቇ㑐ㅪ੐㗄ߩ๮ฬߪ IUPAB ೙ቯߩ๮ฬᴺߦ
ᓥ߁ߎߣ‫ޕ‬㧟㧕ൻቇᑼߦ↪޿ࠄࠇࠆ⸥ภ‫⇛ޔ‬ภߥߤߪ Chemical
Abstracts ߩ↪଀ߦᓥ߁ߎߣ‫ޕ‬㧔㧣㧕⺰ᢥฃℂᓟ‫ޔ‬ቢᚑߒߚᧄᢥ߅ࠃ߮࿑⴫╬ߩශ೚ߦᔅⷐߥోߡߩࡈࠔࠗ࡞ࠍ෼߼ߚ㔚ሶᇦ૕ࠍ✬㓸ᆔຬ㐳ተߦㇷㅍߔࠆ߆‫ࠍ࡞ࠗࠔࡈߩࠄࠇߘߪߚ߹ޔ‬e-mail ᷝઃᦠ㘃ߣߒߡ✬㓸ᆔຬ㐳ተߦㅍઃߔࠆߎߣ‫ޕ‬㧞㧚ᛩⓂ⺰ᢥߩ᭴ᚑ
㧔㧝㧕ේ⪺⺰ᢥߪ 1)ᮡ㗴‫ޔ‬2)ࠕࡉࠬ࠻࡜ࠢ࠻‫ޔ‬3)ᧄ
ᢥ‫ޔ‬4)ᒁ↪ᢥ₂‫ޔ‬5)⴫ߥࠄ߮ߦ࿑‫ޔ‬࿑ߩ⺑᣿ᢥ‫ߩޔ‬㗅ߦ߹ߣ߼ࠆߎߣ‫ޕ‬㧔㧞㧕ᮡ㗴ߪ⺰ᢥౝኈࠍౕ૕⊛ߦ⴫ߔ◲ẖߥ߽ߩߣߔࠆߎߣ‫ޕ‬ᮡ㗴ߩਅߦ⪺⠪ฬ‫ޔ‬ᚲዻᯏ㑐ฬ‫ޔ‬ᚲ࿷࿾ࠍᦠߊ‫⪺ޕ‬⠪߇ⶄᢙߢ‫ޔ‬ᚲዻᯏ㑐߇⇣ߥࠆ႐วߪ‫ޔ‬⪺⠪ฬᧃየߦ਄ઃᢙሼࠍઃߌߡ඙೎ߔࠆߣߣ߽ߦ‫ޔ‬ㅪ⛊ኻᔕ⪺⠪ฬࠍᜰቯߔࠆߎߣ‫ޕ‬㧔㧟㧕ේ⪺⺰ᢥߥࠄ߮ߦ✚⺑ߩࠕࡉࠬ࠻࡜ࠢ࠻ߪ⧷
⺆ߢ⸥ㅀߔࠆ߽ߩߣߒ‫ޔ‬ㅢᏱ⺰ᢥ࡮✚⺑ߢߪ
200words એౝ‫ޔ‬⍴⺰ᢥߢߪ 100words એౝߢ૞ᚑߔ
ࠆߎߣ߇ᦸ߹ߒ޿‫ޕ‬109㧔㧠㧕ㅢᏱ⺰ᢥߩᧄᢥߪේೣߣߒߡ‫᧚ޔ⸒✜ޔ‬ᢱ߅
ࠃ߮ᣇᴺ‫⚿ޔ‬ᨐ‫ޔ‬⠨ኤ㧔޽ࠆ޿ߪ⚿ᨐߣ⠨ኤ㧕‫⻢ޔ‬ㄉߢ᭴ᚑߔࠆ‫ޕ‬⍴⺰ᢥߢߪ㗄⋡ಽߌࠍߒߥ޿‫ޕ‬㧔㧡㧕ේ⪺⺰ᢥߥࠄ߮ߦ✚⺑ߢߪ‫ޔ‬ᔅⷐߦᔕߓߡ࿑
⴫ࠍ↪޿ࠆ‫ߩࠇߙࠇߘޕ‬࿑⴫ࠍᝌ౉ߒߚ޿૏⟎ࠍ‫ޔ‬
ේⓂฝ஥ߩ૛⊕ㇱಽߦᜰ␜ߔࠆߎߣ‫ޕ‬㧔㧢㧕ᢥਛߢߩᢥ₂ߩᒁ↪ߪ‫ޔ‬ᒁ↪ᢥ₂ߩ㗄⋡ߢᢛ
ℂߒߚᢥ₂⇟ภࠍ↪޿ࠆߎߣ‫ޕ‬࿑⴫ߩᒁ↪ߪߘߩ⇟ภߦࠃࠆߎߣ‫ޕ‬㧔㧣㧕⣉ᵈ߇ᔅⷐߥ႐วߦߪ‫⹥ޔ‬ᒰ੐㗄ߩฝ⢋ߦㅢߒ⇟ภࠍઃߌ‫ޔ‬ᒰ⹥㗁ߩਅㇱߦ⺑᣿ᢥࠍઃߌࠆߎߣ‫ޕ‬㧟㧚ᒁ↪ᢥ₂㧔㧝㧕ᒁ↪ᢥ₂ߪਅ⸥ߩ଀ߦḰߓ‫ᧄޔ‬ᢥਛߩ⹥ᒰੱ
ฬ޽ࠆ޿ߪ੐㗄ߩฝ⢋ߦ 1), 1-3)ߩࠃ߁ߦ⇟ภࠍઃߒ‫ᧄޔߚ߹ޔ‬ᢥᧃየߩᒁ↪ᢥ₂ߩ㗄ߦ╙৻⪺⠪ߩࡈࠔࡒ࡝࡯ࡀ࡯ࡓߩࠕ࡞ࡈࠔࡌ࠶࠻㗅ߦ 1., 2., 3., --ߩ⇟ภࠍઃߒ৻᜝⸥タߔࠆ‫ޕ‬Ibid., idem ߪ↪޿ߥ޿ߎߣ‫⑳ޕ‬ା‫⎇ߩ⴫⊒ᧂޔ‬ⓥ⚿ᨐ‫ޔ‬ቇⴚ㔀⹹ߦฃℂߐࠇࠆ೨ߩ⺰ᢥ‫ޔ‬ᛞ㍳߇ශ೚ߐࠇߡ޿ߥ޿ญ㗡⊒⴫ߥߤߪᒁ↪ᢥ₂ߦ฽߼ߥ޿ߎߣ‫ޕ‬㔀⹹ߩ⇛ภߪ‫ޔ‬Chemical
Abstracts Service Source Index (CASSI) 1907-1984(Cumulative)߅ࠃ߮ߘߩ⵬ㆮ ߦࠃࠅ‫ޔ‬㔀⹹ฬ‫ᦠޔ‬ฬ‫ޔ‬ᐕ‫ޔ‬Ꮞ߅ࠃ߮㗁ߩ␜ߒᣇߪਅ⸥ߦࠃࠆߎߣ‫ޕ‬㧔㧞㧕㔀⹹ᒁ↪ߩߣ߈
Chuakrut, S., Arai, H., Ishii, M., and Igarashi, Y. 2000.
Characterization of a bifunctional archaeral acylcoenzyme A carboxylase. J. Bacteriol. 185: 938~947.
㧔㧟㧕ᦠ☋ᒁ↪ߩߣ߈ఽ₹ᔀ. 1997. V ㄭઍ⊛ߥᓸ↢‛೑↪Ꮏᬺ㧝㧚ࠕ࡞ࠦ࡯࡞, pp191~197.ఽ₹ᔀ࡮ᾢ⼱⧷ᒾ✬, 㘩ຠᓸ↢‛ቇ, ᢥ᳗ၴ಴
Kato, C. 2011. 5.2 High pressure and prokaryotes. In:“Extremophiles Handbook” pp. 657-668. (Eds.
Horikoshi, K., Antranikian, G., Bull, A., Robb, F. and
Stetter, K.), Springer-Verlag, Tokyo.
㧠㧚⴫ߣ࿑߅ࠃ߮ߘߩ⺑᣿㧔㧝㧕⴫ߣ࿑ߪශ೚ ਅࠍ૞ᚑߔࠆߎߣ‫ޕ‬㧔㧞㧕ᧄᢥࠍ⧷⺆ߢᦠ޿ߚ႐วߦߪ࿑⴫߽⧷ᢥߢ૞
ᚑߔࠆߎߣ‫ޕ‬㧔㧟㧕⴫ߣ࿑ߪ৻ߟߦኻߒߡ㧝㗁૶߁‫ޕ‬ฦ㗁ߩฝ਄ߦ⪺⠪ฬߥࠄ߮ߦ࿑ߩ⇟ภࠍ౉ࠇࠆߎߣ‫ޕ‬㧔㧠㧕⴫㗴ߪ⴫ߩ਄ㇱಽߦᦠߊߎߣ‫ߩ⴫ޕ‬ౝኈ⺑᣿ᢥߥࠄ߮ߦᵈߪ‫ߩ⴫ޔ‬ਅߦ⸥ㅀߔࠆ‫ޕ‬ᵈߩ⴫␜ߦߪ
a),b),c)ࠍ਄ઃߢ᣿⸥ߒ‫⸥ޔ‬ㅀߦߪ a),b),c)ࠍ਄ઃߦߖ
ߕ↪޿ࠆ‫ޕ‬㧔㧡㧕࿑ߩᮡ㗴ߥࠄ߮ߦ⺑᣿ᢥߪ೎⚕ߦ߹ߣ߼ߡශ
೚ߔࠆߎߣ‫ޕ‬࿑ߩᮡ㗴ߩᦨᓟߦߪࡇ࡝ࠝ࠼ࠍઃߒ‫ޔ‬ౝኈ⺑᣿ߪᮡ㗴ߩᓟߦᡷⴕߒߡ⸥タߔࠆߎߣ‫ޕ‬㧔㧢㧕౮⌀ߪ᣿⍎ߥ㓁↹ࠍᔅⷐㇱᢙᷝઃߔࠆߎߣ‫ޕ‬

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)ቇળળೣᭂ㒢ⅣႺ↢‛ቇળળೣ㧔ฬ⒓㧕╙㧝᧦ ᧄળߪ‫ޔ‬ᭂ㒢ⅣႺ↢‛ቇળ㧔⧷ᢥฬ㧦The
Japanese Society for Extremophiles 㧕ߣ޿߁‫ޕ‬㧔એਅᧄળߣ޿߁㧕㧔⋡⊛㧕╙㧞᧦ ᧄળߪ‫ޔ‬ᭂ㒢ⅣႺ↢‛⎇ⓥߩ⊒ዷߦነਈߒ‫ޔ‬ᭂ㒢ⅣႺ↢‛ߩ⎇ⓥ‫ޔ‬ᢎ⢒‫ޔ‬೑↪ߩផㅴߦነਈߔࠆߎߣࠍ⋡⊛ߣߔࠆ‫ޕ‬㧔੐ᬺ㧕╙㧟᧦ ᧄળߪ‫⎇ޔ‬ⓥ⊒⴫ળ෸߮⻠Ṷળ╬ߩ㐿௅‫ޔ‬ᭂ 㒢 Ⅳ Ⴚ ↢ ‛ ቇ ળ ⹹ 㧔 ⧷ ฬ 㧦 Journal ofJapanese Society for Extremophiles㧕ߩ⊒ⴕ‫ޔ‬࿖㓙ᭂ㒢ⅣႺ↢‛ቇળ╬ߣߩㅪ៤੐ᬺ‫⎇ޔ‬ⓥᅑബ⾨࡮ࡐࠬ࠲࡯⾨ߩ᝼ਈ‫ઁߩߘޔ‬೨᧦ߩ⋡⊛ࠍ㆐ᚑߔࠆߚ߼ߦᔅⷐߥ੐ᬺࠍⴕ߁‫ޕ‬㧔ળຬߩ⒳೎╬㧕╙㧠᧦
1. ᧄળߩળຬߪᱜળຬ‫ޔ‬ฬ⹷ળຬ‫ޔ‬࿅૕ળຬ߅ࠃ߮⾥ഥળຬߣߔࠆ‫ޕ‬
2. ᱜળຬߪᭂ㒢ⅣႺ↢‛ߦ㑐ߔࠆ⎇ⓥߦᓥ੐ߔࠆ‫ߦࠇߎޔߪߚ߹ޔ‬㑐ᔃࠍᜬߟ୘ੱߢ޽
ߞߡ‫ᧄޔ‬ળߩ⋡⊛ߦ⾥หߒ‫ޔ‬ቯ߼ࠄࠇߚળ⾌ࠍ⚊߼ߚ⠪ࠍ޿߁‫ޕ‬
3. ฬ⹷ળຬߪ‫ޔ‬ᒰቇળߩ⊒ዷ߿ಽ㊁ߦഞഭߩ޽ߞߚ⠪ߢ‫ޔ‬ᐙ੐ળߦࠃࠆផ⮈‫⼏⹏ޔ‬ຬળߦࠃࠆᛚ⹺ࠍ⚻ߡ‫ޔ‬ળ㐳ߩᜰฬߦࠃࠅ᳿ቯߐࠇ‫ޔ‬ᒰቇળߩ௅ߔࠪࡦࡐࠫ࠙ࡓ࡮ᐕળ╬ߦ᜗ᓙߐࠇࠆ‫ޕ‬
4. ࿅૕ળຬߪᭂ㒢ⅣႺ↢‛ߦ㑐ᔃࠍᜬߟ࿅
૕㧔ᴺੱ╬㧕ߢ޽ߞߡ‫ߩߘޔ‬࿅૕ౝߩᧄળߩ
⋡⊛ߦ⾥หߔࠆઍ⴫⠪ࠍ฽߼ߡ㧟ฬએౝߩᚲዻ⠪ࠍᜰฬߒߡળຬ⊓㍳ࠍⴕ޿‫ޔ‬ቯ߼ࠄࠇߚળ⾌ࠍ⚊߼ߚ࿅૕ࠍ޿߁‫ޕ‬
5. ⾥ഥળຬߪᧄળߩ⋡⊛ߦ⾥หߒ‫ޔ‬ቯ߼ࠄࠇߚ⾥ഥળ⾌ࠍ㧝ญએ਄⚊߼ߚ୘ੱ߹ߚߪ࿅
૕ࠍ޿߁‫ޕ‬╙㧡᧦ ળຬߪᧄળ߇㐿௅ߔࠆ⻉੐ᬺߦෳടߒ‫ᧄޔ‬ળߩ⊒ⴕߔࠆශ೚‛‫ޔ‬PDF ╬ߩ㈩Ꮣࠍฃߌ
ࠆߎߣ߇ߢ߈ࠆ‫ޕ‬㧔౉ળ㧕╙㧢᧦ ળຬߣߒߡ౉ળߒࠃ߁ߣߔࠆ୘ੱ߹ߚߪ࿅
૕ߪ‫ᧄޔ‬ળ੐ോዪ߇ቯ߼ࠆᚻ⛯߈ߦᓥߞߡ
↳ߒㄟߺࠍⴕ޿‫ᧄޔ‬ળળ㐳ߦࠃࠆᛚ⹺ࠍᓧ
ߥߌࠇ߫ߥࠄߥ޿‫ޕ‬

110

㧔ળ⾌㧕
╙㧣᧦ ળຬߪਅ⸥ߩળ⾌ࠍ⚊߼ࠆ߽ߩߣߔࠆ‫ޕ‬ᱜળຬ  ᐕ㗵 5,000 ౞㧔ૉߒりಽ߇ቇ↢ߢ޽ࠆ⠪ߪᐕ㗵 2,000 ౞㧕ฬ⹷ળຬ ᐕ㗵 0 ౞࿅૕ળຬ ᐕ㗵 20,000 ౞⾥ഥળຬ ᐕ㗵 ৻ญએ਄㧔৻ญ 30,000 ౞㧕㧔ㅌળ㧕╙㧤᧦ ળຬߪળ㐳ߦዯߌࠆߎߣߦࠃߞߡㅌળߔࠆ
ߎߣ߇ߢ߈ࠆ‫ޕ‬㧔ᓎຬ㧕╙㧥᧦
1. ᧄળߦߪળ㐳 1 ฬ‫ޔ‬೽ળ㐳 2 ฬ‫⼏⹏ޔ‬ຬ⧯ᐓฬ‫ޔ‬ᐙ੐⧯ᐓฬ‫ޔ‬ળ⸘⋙ᩏ 2 ฬߩᓎຬࠍ߅ߊ‫ޕ‬
2. ળ㐳ߪᧄળࠍઍ⴫ߒ‫ޔ‬ળോࠍ⛔᜝ߔࠆ‫ޕ‬
3. ೽ળ㐳ߪળ㐳ࠍ⵬૒ߒ‫ޔ‬ળ㐳ߦ੐᡿޽ࠆ႐
วߦߪળ㐳ߩ⡯ോࠍઍⴕߔࠆ‫ޕ‬
4. ᐙ੐ߪ‫ޔ‬ᐙ੐㐳‫ޔ‬ᐼോ‫ޔ‬ቇⴚᵴേ‫ޔ‬ળ⸘‫ޔ‬ࠍಽᜂߒ‫ޔ‬੐ᬺ⸘↹ߩ┙᩺‫ޔ‬੐ᬺߩታᣉ‫ޔ‬ᐼോળ⸘‫ߩࡓ࠙ࠫࡐࡦࠪޔ‬ડ↹┙᩺‫ޔ‬ቇળ⹹⊒ⴕ‫ޔ‬࿖㓙ᭂ㒢ⅣႺ↢‛ቇળ╬ߣߩㅪ⛊ߥߤߩᧄળߩ੐ോࠍⴕ߁‫ޕ‬
5. ળ⸘⋙ᩏߪᧄળߩળ⸘ࠍ⋙ᩏߔࠆ‫ޕ‬㧔ᓎຬߩㆬ಴㧕╙㧝㧜᧦ ળ㐳‫ޔ‬೽ળ㐳‫⼏⹏ޔ‬ຬߪᧄળߩ✚ળߦ߅޿ߡㆬ಴ߐࠇࠆ‫ޕ‬
1. ᐙ੐‫ޔ‬ળ⸘⋙ᩏߪ‫ޔ‬೽ળ㐳‫⼏⹏ޔ‬ຬએᄖߩᱜળຬߩਛ߆ࠄળ㐳߇ᜰฬߒ‫✚ޔ‬ળߩᛚ⹺ࠍฃߌࠆ‫ޕ‬2. ⹏⼏ຬߩછᦼߪ㧞ᐕߣߒ㊀છࠍᅹߍߥ޿
߇‫ޔ‬ㅪ⛯㧟ᦼࠍ㒢ᐲߣߔࠆ‫ޕ‬ૉߒ‫ޔ‬ળ㐳߇ᜰฬߔࠆ⹏⼏ຬߪߎߩ㒢ࠅߢߪߥ޿‫ޕ‬㧔ᆔຬળ㧕╙㧝㧝᧦ ᧄળߦቇળ⹹✬㓸ᆔຬળ‫⎇ޔ‬ⓥᅑബ⾨ㆬ⠨ᆔຬળ‫ࡓ࠙ࠫࡐࡦࠪޔ‬ᆔຬળࠍ⟎ߊ‫ޕ‬ฦᆔຬળᆔຬߩછᦼߪ 2 ᐕߣߔࠆ߇㊀છࠍߐ߹ߚߍߥ޿‫ޕ‬
1. ቇળ⹹✬㓸ᆔຬળ㧔ળ㐳ᜰฬߩ⧯ᐓฬߢ
᭴ᚑߒ㨮ᆔຬ㐳ߪ੕ㆬߔࠆ㧕ߪቇળ⹹ߩ✬㓸‫ޔ‬⺰ᢥክᩏߩછߦ޽ߚࠆ‫ޕ‬
2. ⎇ⓥᅑബ⾨ㆬ⠨ᆔຬળ㧔೽ળ㐳߇ᆔຬ㐳ࠍൕ߼‫ޔ‬ቇⴚᵴേᜂᒰᐙ੐ߣᆔຬ㐳ᜰฬߩᆔຬ‫ ⸘ޔ‬10 ฬ⒟ᐲߢ᭴ᚑߔࠆ㧕ߪᧄળߩ⎇ⓥ
ᅑബ⾨ฃ⾨⠪ࠍㆬ⠨ߔࠆ‫ޕ‬
3. ࠪࡦࡐࠫ࠙ࡓᆔຬળ㧔ળ㐳ᜰฬߩ⧯ᐓฬ

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)㧔ઃೣ㧕╙㧝㧢᧦ ᧄળೣߩᣉⴕ߅ࠃ߮ᧄળߩㆇ༡ߦߟ޿
ߡߩ⚦ೣߪ✚ળߩ⼏᳿ࠍ⚻ߡ೎ߦቯ߼ࠆ‫ޕ‬╙㧝㧣᧦ ᧄળೣߩᄌᦝߪ✚ળߩ⼏᳿ࠍ⚻ߡⴕ߁‫ޕ‬╙㧝㧤᧦ ᧄળೣߪ‫ޔ‬1999 ᐕ 10 ᦬ 19 ᣣࠃࠅᣉⴕߔ
ࠆ‫ޕ‬ߢ᭴ᚑߒ㨮ᆔຬ㐳ߪ੕ㆬߔࠆ㧕ߪቇⴚᵴേᐙ
੐ߣ౒หߢࠪࡦࡐࠫ࠙ࡓࠍડ↹┙᩺ߒ‫ޔ‬ㆇ༡ࠍࠨࡐ࡯࠻ߔࠆ‫ޕ‬㧔⹏⼏ຬળ㧕
╙㧝㧞᧦ ⹏⼏ຬળߪળ㐳‫ޔ‬೽ળ㐳‫⼏⹏ޔ‬ຬࠍ߽ߞߡ᭴ᚑߔࠆ‫ޕ‬ળ㐳ߪᐕ㧝࿁એ਄⹏⼏ຬળࠍถ㓸ߒ‫⼏ޔ‬㐳ߣߥࠅ‫ޔ‬ળߩ㊀ⷐ੐㗄ࠍክ⼏ߒ✚ળߦឭ᩺ߔࠆ‫ޕ‬㧔✚ળ㧕╙㧝㧟᧦ ᧄળߪේೣߣߒߡᐕ㧝࿁✚ળࠍ㐿߈‫ޔ‬ળോࠍද⼏ߒ⼏᳿ߔࠆ‫ޕ‬✚ળߪળ㐳߇ถ㓸ߔࠆ‫ޕ‬╙㧝㧠᧦ ✚ળߩ⼏᳿ߪ಴Ꮸળຬߩㆊඨᢙߩ⾥ᚑࠍ߽ߞߡⴕ߁‫ޕ‬㧔ળ⸘ᐕᐲ㧕
╙㧝㧡᧦ ᧄળߩ੐ᬺᐕᐲߪ 1 ᦬ 1 ᣣߦᆎ߹ࠅ‫ޔ‬หᐕ 12 ᦬ 31 ᣣߦ⚳ࠆ‫ޕ‬ቇળ⚦ೣ

2001 ᐕ 11 ᦬ 29 ᣣᡷቯ
2002 ᐕ 12 ᦬ 1 ᣣᡷቯ
2003 ᐕ 12 ᦬ 1 ᣣᡷቯ2005 ᐕ 11 ᦬ 7 ᣣᡷቯ2007 ᐕ 12 ᦬ 1 ᣣᡷቯ2010 ᐕ 1 ᦬ 1 ᣣᡷቯ2012 ᐕ 12 ᦬ 1 ᣣᡷቯ      ᭂ㒢ⅣႺ↢‛ቇળ⚦ೣ╙㧤᧦ ⹏⼏ຬળߩ᳿⼏ߪ಴Ꮸ⠪ߩㆊඨᢙߩ⾥ᚑߦࠃࠅᚑ┙ߔࠆ‫ޕ‬㧔ㅌળ㧕╙㧥᧦ ળ㐳ߪ‫ޔ‬ળ⾌ࠍ㧟ᐕએ਄ṛ⚊ߒߚળຬࠍㅌ
ળߐߖࠆߎߣ߇಴᧪ࠆ‫ޕ‬㧔᝼⾨㧕╙㧝㧜᧦ ⎇ⓥᅑബ⾨࡮ࡐࠬ࠲࡯⾨ߩ᝼⾨ⷙቯߪ೎ㅜቯ߼ోળຬߦ㈩Ꮣߔࠆ‫ޕ‬㧔⚦ೣߩᄌᦝ㧕╙㧝㧝᧦ ᧄ⚦ೣߩᄌᦝߪ✚ળߩ⼏᳿ߦࠃࠆ‫ޕ‬㧔ઃೣ㧕╙㧝㧞᧦ ੹ᦼߩᧄળ੐ോዪߪ‫᧲ޔ‬੩Ꮏᬺᄢቇᄢቇ
㒮↢๮ℂᎿቇ⎇ⓥ⑼ ਛ᧛⡡⎇ⓥቶ㧔ᮮᵿᏒ✛඙㐳ᵤ↰↸㧕ౝߣߔࠆ‫ޕ‬╙㧝㧟᧦ ᧄ⚦ೣߪ‫ޔ‬1999 ᐕ 10 ᦬ 19 ᣣࠃࠅߎࠇࠍ
ታᣉߔࠆ‫ޕ‬㧔ળຬ㧕╙㧝᧦ ౉ળࠍᛚ⹺ߐࠇߚᱜળຬ‫ޔ‬ฬ⹷ળຬ‫ޔ‬࿅૕ળຬ‫⾥ޔ‬ഥળຬߪ‫ޔ‬ᚲቯߩળ⾌ࠍㅦ߿߆ߦ
⚊౉ߔࠆ‫ޕ‬㧔✚ળ㧕╙㧞᧦ ✚ળߩ⼏᩺ߪ೽ળ㐳‫ޔ‬ᐙ੐╬ߣ౒ߦળ㐳߇૞ᚑߒ‫⼏⹏ޔ‬ຬળߩ⼏ࠍ⚻ߚᓟឭ಴ߔࠆ‫ޕ‬⼏᩺ߦߪ೨ᐕᐲߩ੐ᬺౝኈ෸߮෼ᡰ᳿▚‫ޔ‬ᣂᐕᐲߩ੐ᬺ⸘↹‫ޔ‬෸߮෼ᡰ੍▚ࠍ฽߻߽ߩߣߔࠆ‫ޕ‬╙㧟᧦ ✚ળߪળຬߩ㧝㧛㧝㧜એ਄ߩ಴Ꮸ㧔ૉߒᆔછ⁁ࠍ฽߻㧕ࠍ߽ߞߡᚑ┙ߔࠆ‫ޕ‬㧔ᓎຬߩㆬ಴㧕╙㧠᧦ ળ㐳‫ޔ‬೽ળ㐳‫⼏⹏ޔ‬ຬߪળຬߩㆬ᜼ߦࠃߞ
ߡ᳿߼ࠄࠇࠆ‫ޕ‬ળ㐳ߪㆬ᜼ߦࠃࠄߕ⹏⼏ຬࠍᜰฬߔࠆߎߣ߇಴᧪ࠆ߇‫✚ޔ‬ળߢߩᛚ⹺
ࠍᔅⷐߣߔࠆ‫ޕ‬╙㧡᧦ ળ㐳ߪળຬߩਛ߆ࠄዋߥߊߣ߽㧟ฬࠍㆬࠎߢㆬ᜼▤ℂᆔຬࠍᆔབྷߔࠆ‫ޕ‬ㆬ᜼▤ℂᆔຬ
ߪㆬ᜼੐ോࠍⴕ߁‫ޕ‬╙㧢᧦ ળ㐳ߪ‫⼏⹏ޔ‬ຬળߩᛚ⹺ࠍᓧߡᔅⷐߥᆔຬ
ળࠍ૞ࠆߎߣ߇಴᧪ࠆ‫ޕ‬㧔⹏⼏ຬળ㧕
╙㧣᧦ ⹏⼏ຬળߪ⹏⼏ຬߩඨᢙએ਄ߩ಴Ꮸ㧔ᆔછ⁁ࠍ฽߻㧕ࠍ߽ߞߡᚑ┙ߔࠆ‫ޕ‬

2002 ᐕ 12 ᦬ 1 ᣣᡷቯ
2003 ᐕ 12 ᦬ 1 ᣣᡷቯ
2006 ᐕ 6 ᦬ 30 ᣣᡷቯ
2007 ᐕ 12 ᦬ 1 ᣣᡷቯ2010 ᐕ 1 ᦬ 1 ᣣᡷቯ2012 ᐕ 12 ᦬ 1 ᣣ111

Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)

ㆇ༡૕೙
ળ㐳㩷
೽ળ㐳

ᭂ㒢ⅣႺ↢‛ቇળㆇ༡૕೙㧔2013 ᐕᐲ㧕ជ⿧ ᒄᲞ㩷⁛┙ⴕ᡽ᴺੱ ᶏᵗ⎇ⓥ㐿⊒ᯏ᭴㩷ᄢፉ ᵏ㇢㩷౒๺ൻᎿᩣᑼળ␠ ⅣႺᓸ↢‛ቇ⎇ⓥᚲ㩷Ꮏ⮮ ବ┨㩷㐳ፒᄢቇ㩷 ᳓↥ቇㇱ㩷⹏⼏ຬ㧔㧡㧜㖸㗅‫ޔ‬ᢘ⒓⇛㧕

ᐙ੐ળ

㩷

㩷

⑺㊁೑㇢

ࠧ࡯࠹࠶ࠢᩣᑼળ␠

ᐙ੐㐳

ਛ᧛⡡

᧲੩Ꮏᬺᄢቇᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼

੖ච፲ᵏᄦ

᧲੩ᄢቇᄢቇ㒮ㄘቇ↢๮⑼ቇ⎇ⓥ⑼

ᐼോદ⮮᡽ඳ

᧲ᵗᄢቇ↢๮⑼ቇㇱ

⍹㊁࿦ሶ

਻Ꮊᄢቇᄢቇ㒮ㄘቇ⎇ⓥ㒮

ቇⴚᵴേ

દ⮮ㅴ

℄⃿ᄢቇㄘቇㇱ✬㓸ᆔຬ㐳

⍹੗ᱜᴦ

᧲੩ᄢቇᄢቇ㒮ㄘቇ↢๮⑼ቇ⎇ⓥ⑼

દ⮮૓ሶ

ഃଔᄢቇᎿቇㇱቇળ⹹ᜂᒰ

⍹㊁⦟⚐

਻Ꮊᄢቇᄢቇ㒮ㄘቇ⎇ⓥ㒮┙๮㙚ᄢቇ↢๮⑼ቇㇱቇળ⹹ᜂᒰ㜞ຠ⍮ౖ

᧲ᵗᄢቇ↢๮⑼ቇㇱᄢፉᢅਭ

਻Ꮊᄢቇᄢቇ㒮ㄘቇ⎇ⓥ㒮ቇળ⹹ᜂᒰฎ࿦ߐ߅ࠅ

᧲੩ᄢቇ↢‛↢↥Ꮏቇ⎇ⓥ࠮ࡦ࠲࡯

㎨ᒻᵗ৻

⁛┙ⴕ᡽ᴺੱ↥ᬺᛛⴚ✚ว⎇ⓥᚲળ⸘ቝ૒⟤⺰

᧲ᵗᄢቇℂᎿቇㇱᴡේᨋ⵨

਻Ꮊᄢቇᄢቇ㒮ㄘቇ⎇ⓥ㒮ળ⸘㆏ਭೣਯ

᧲ᵗᄢቇ↢๮⑼ቇㇱୖశᚑ♿

ᄢ㒋ᄢቇᄢቇ㒮ℂቇ⎇ⓥ⑼

HP ᜂᒰਃベື਽

⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ዊᨋືᄦ

ฬฎደᄢቇᄢቇ㒮↢๮ㄘቇ⎇ⓥ⑼

HP ᜂᒰὑᚒ੗⑲ⴕ

ᣣᧄᄢቇᢥℂቇㇱዊᨋ⦟ೣ

⽷࿅ᴺੱࡃࠗࠝࠗࡦ࠳ࠬ࠻࡝࡯දળ

ࠪࡦࡐࠫ࠙᧲੩Ꮏᬺᄢቇᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼

᧲ർᄢቇ෻ᔕൻቇ⎇ⓥᚲࡓᜂᒰ౎ᵄ೑ᕺ

૒਄ඳ
⦼ᒄቁ

ࠨࡦ࠻࡝࡯ࡆࠫࡀࠬࠛࠠࠬࡄ࡯࠻ᩣᑼળ␠

ቇⴚᐙ੐〔⷗᥍ᐘ

੩ㇺᄢቇᄢቇ㒮Ꮏቇ⎇ⓥ⑼

⩲ේ⑲᣿

࿖┙ㆮવቇ⎇ⓥᚲ

ቇⴚᐙ੐

દ⮮㓉

⁛┙ⴕ᡽ᴺੱℂൻቇ⎇ⓥᚲ

㋈ᧁஜ৻ᦶ

⁛┙ⴕ᡽ᴺੱ⵾ຠ⹏ଔᛛⴚၮ⋚ᯏ᭴ቇⴚᐙ੐ખቬᩮ⮍

ㄭ⇰ᄢቇᎿቇㇱਛᎹ๺୶

ᗲᇫ⋵┙੹ᴦ⷏㜞╬ቇᩞቇⴚᐙ੐࿡ᨑᱞ๺

᧲੩ᄢቇᄢቇ㒮ℂቇ♽⎇ⓥ⑼

㐳↰ᵗሶ

ᣣᧄᄢቇℂᎿቇㇱ  ቇⴚᐙ੐㕍ᧁବᄦ

ᣣᧄᄢቇ↢‛⾗Ḯ⑼ቇㇱධㇱṑ㇢

ᣣᧄ⿒චሼ਻Ꮊ࿖㓙⋴⼔ᄢቇ⋴⼔ቇㇱቇⴚᐙ੐ᩙේ㆐ᄦ

੩ㇺᄢቇൻቇ⎇ⓥᚲ

㐳⼱Ꮉഞ

ᣣᧄᄢቇ↢‛⾗Ḯ⑼ቇㇱቇⴚᐙ੐⑔ዬବᤘ

᧲੩Ꮏᬺᄢቇᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼

ඨỈᢅ᧲࠰࡯ᩣᑼળ␠᧲੩⎇ⓥᚲ 

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᧲ᵗᄢቇ↢๮⑼ቇㇱ೎ᐭノᒾ

ᣣᧄᄢቇ✚ว⑼ቇ⎇ⓥᚲቇⴚᐙ੐੉ୖᱜඳ

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᧻੗⵨⍹Ꮉ⋵┙ᄢቇ↢‛⾗ḮᎿቇ⎇ⓥᚲቇⴚᐙ੐ട⮮ජ᣿

⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ਣጊᱜ⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ળ⸘⋙ᩏ㜞ἑశᓼ

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᧲੩ᄢቇᄢቇ㒮ㄘቇ↢๮⑼ቇ⎇ⓥ⑼

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᧲੩⮎⑼ᄢቇ↢๮⑼ቇㇱࠪࡦࡐࠫ࠙ࡓᆔຬળ

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᧲ᵗᄢቇ ↢๮⑼ቇㇱ   ᆔຬખጊ ⧷᮸

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ජ⪲ᄢቇ ࿦⧓ቇㇱ 㤛Ꮉ↰ 㓉ᵗ⁛┙ⴕ᡽ᴺੱㄘᬺ↢‛⾗Ḯ⎇ⓥᚲ ፃጯ ብ᣿

᧲ᵗᄢቇ ࡃࠗࠝ࠽ࡁࠛ࡟ࠢ࠻ࡠ࠾ࠢࠬ
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ർ⷗Ꮏᬺᄢቇ Ꮏቇㇱ ᛬↰ ๺ᴰ᧲੩Ꮏᬺᄢቇᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼Journal of Japanese Society for Extremophiles (2012) Vol.11 (2)✬ 㓸 ᓟ ⸥੹ᐕᐲߩᐕળߪ᧲੩਎↰⼱ߩᣣᧄᄢቇᢥℂቇㇱߩ࿾ߢ㐿௅ߐࠇ߹ߒߚ‫ޕ‬਎↰⼱ߣ޿߁࿾ߪ‫ޔ‬㜞⚖૑ቛⴝ߇┙ߜਗ߱ࠗࡔ࡯ࠫ߇޽ࠅ߹ߔ߇‫ޔߪߎߎޔ‬᪉ਗᧁ߇⛯ߊߣߡ߽㕒߆ߥ႐ᚲߢ߽޽ࠅ߹ߔ‫ޕ‬ᤐߩ᪉ߩ㐿⧎ߩቄ▵ߦߥࠅ߹ߔߣ‫ޔ‬᪉ߩฬᚲߣߒߡ߅⧎⷗ߦ⸰ࠇࠆ
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߅㗿޿↳ߒ਄ߍ߹ߔ‫ޕ‬ቇળ⹹✬㓸ᜂᒰᐙ੐ ട⮮ජ᣿✬㓸ᆔຬળᆔຬ㐳 㧦⍹੗ᱜᴦ㧔᧲ᄢ㧕೽ᆔຬ㐳㧦⍹㊁⦟⚐㧔਻ᄢ㧕ᆔ  ຬ㧦㒙ㇱᢥᔟ㧔㕍ቇᄢ㧕‫ޔ‬દ⮮ 㓉㧔ℂ⎇㧕‫ޔ‬㎨ᒻᵗ৻㧔↥✚⎇㧕‫ޔ‬ᴡේᨋ⵨㧔਻Ꮊᄢ㧕‫ޔ‬㜞ຠ⍮ౖ㧔᧲ᵗᄢ㧕‫ޔ‬ὑᚒ੗⑲ⴕ㧔ᣣᄢ㧕‫ޔ‬ਛ᧛ ⡡㧔᧲Ꮏᄢ㧕‫ޔ‬ਛ᧛ 㗼㧔╳ᵄᄢ㧕‫ޔ‬ਃベື਽㧔JAMSTEC㧕‫ޔ‬ḡᧄ ൟ㧔↥✚⎇㧕ቇળ੐ോዪ
‫ޥ‬226-8501 ᮮᵿᏒ✛඙㐳ᵤ↰↸ 4259-J2-13᧲੩Ꮏᬺᄢቇ ᄢቇ㒮↢๮ℂᎿቇ⎇ⓥ⑼↢‛ࡊࡠ࠮ࠬኾ᡹ ਛ᧛⡡⎇ⓥቶౝᭂ㒢ⅣႺ↢‛ቇળ੐ോዪ ਛ᧛ ⡡ (ᐙ੐ળᐙ੐㐳)
FAX: 045-924-5837 E-mail: kyokugenjm10@extremophiles.jpቇળ⹹✬㓸‫ޥ‬237-0061 ␹ᄹᎹ⋵ᮮ㗇⾐Ꮢᄐፉ↸ 2-15⁛┙ⴕ᡽ᴺੱᶏᵗ⎇ⓥ㐿⊒ᯏ᭴ ᶏᵗ࡮ᭂ㒢ⅣႺ↢‛࿤㗔ၞ (ട⮮ජ᣿‫ޔ‬ቇⴚᐙ੐)TEL㧦046-867-9550 FAX㧦046-867-9525 E-Mail㧦kato_chi@jamstec.go.jp⊒ೀᣣ 2012 ᐕ 12 ᦬ 31 ᣣ

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