ㄽᩥᑂᰝࡢせ᪨༤ኈࡢᑓᨷศ㔝ࡢྡ⛠༤ ኈ㸦 ⌮Ꮫ 㸧Ꮫ఩ᤵ୚ࡢせ௳Ꮫ఩つ๎➨ 4 ᮲➨ 1࣭2 㡯ヱᙜẶྡ   㧘 ᶫ ྐ ဨㄽᩥ㢟┠

Study on long-chain alcohol utilization pathways in the glycolipid-producing yeast
Starmerella bombicola㸦⢾⬡㉁⏕⏘⳦ Starmerella bombicola ࡟࠾ࡅࡿ㛗㙐࢔ࣝࢥ࣮ࣝ㈨໬⤒㊰࡟㛵ࡍࡿ◊✲㸧ㄽᩥᑂᰝᢸᙜ⪅୺  ᰝ   ᩍ ᤵ  ᒣ ᮏ༟ᑂᰝጤဨ   ᩍ ᤵ  ஭ ฟ༤                ᑂᰝጤဨ   ᩍ ᤵ  ᆏ ᮏᩔ                ᑂᰝጤဨ   ᩍ ᤵ  Ἠ㍜ಇᑂᰝጤဨ   ㅮ ᖌ  బஂ㛫 ဴྐ                ࢔࣏ࣝ࢟ࣝࣜࢢࣜࢥࢩࢻ㸦APGs㸧ࡣࠊ㧗࠸㉳Ἳᛶ࡜⫙࡬ࡢ㠀౵くᛶࢆేࡏᣢࡘᏳᚰ࣭Ᏻ඲࡞ඃࢀࡓὙί๣࡛࠶ࡾࠊྛ✀ࡢὙ๣➼࡟㓄ྜࡉࢀ࡚࠸ࡿࠋAPGsࡢᕤᴗⓗ⏕⏘ࡣࠊ୺࡟⢾࡜࢔ࣝࢥ࣮ࣝࢆཎᩱ࡜ࡋࡓ໬Ꮫྜᡂ࡟ࡼࡾ⾜ࢃࢀ࡚࠸ࡿࡀࠊཎᩱ࢔ࣝࢥ࣮ࣝࢆከ㔞࡟ᚲせ࡜ࡋࠊ㧗࣭㧗ᅽୗ࡛ࡢ཯ᛂࢆ⾜࠺ࡓࡵ࡟ཎᩱࢢࣝࢥ࣮ࢫࡀኚᛶࡋ࡚ࡋࡲ࠺࡞࡝ࡢၥ㢟ࡀ࠶ࡿࠋࡇࡢࡼ࠺࡞≧ἣ࠿ࡽࠊࡼࡾຠ⋡ࡢ㧗࠸⏕⏘᪉ἲࢆ㛤Ⓨࡍࡿࡓࡵࠊᚤ⏕≀ࢆ⏝࠸ࡓᖖࠊᖖᅽ࡛ࡢAPGsࡢ〇㐀ᢏ⾡ࡀ᳨ウࡉࢀ࡚ࡁࡓࠋࣂ࢖࢜ࢧ࣮ࣇ࢓ࢡࢱࣥࢺ࡜ࡋ࡚᭷⏝࡞ࢯ࣍ࣟࣜࣆࢵࢻࡢ⏕⏘㓝ẕ࡛࠶ࡿStarmerella bombicolaࢆ⢾࡜2⣭࢔ࣝࢥ࣮ࣝࢆྵ᭷ࡍࡿᇵᆅ࡛ᇵ㣴ࡍࡿ࡜ࠊ⏕⏘≀࡜ࡋ࡚APGsࡢ୍✀࡛࠶ࡿ࢔ࣝ࢟ࣝࢯ࣍ࣟࢩࢻࢆ⏕ᡂࡍࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋࡋ࠿ࡋ࡞ࡀࡽࠊᏳ౯࡞1⣭࢔ࣝࢥ࣮ࣝࢆཎᩱ࡜ࡋࡓሙྜࠊ⏕⏘≀཰⋡ࡀⴭࡋࡃῶᑡࡍࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋࡑࡇ࡛ᮏ◊✲࡛ࡣࠊᚤ⏕≀࡟ࡼࡿᖖ࣭ᖖᅽୗ࡛ࡢ㧗཰⋡࡞APGs〇㐀ἲࢆᵓ⠏ࡍࡿࡓࡵࠊࡇࢀࡲ࡛࡟୙࡛᫂࠶ࡗࡓS. bombicolaࡢ࢔ࣝࢥ࣮
ࣝ㈨໬⤒㊰ࡢゎ᫂ࢆ⾜࠺ࡇ࡜ࢆ┠ⓗ࡜ࡋࡓࠋ ➨㸯❶࡛ࡣࠊS. bombicola ࡟࠾ࡅࡿ 1 ⣭࢔ࣝࢥ࣮ࣝ㈨໬⤒㊰ࡢゎ᫂࡜ࠊ㛗㙐࢔ࣝࢥ࣮ࣝ㓟໬㓝⣲ FAO1 㑇ఏᏊ◚ቯᰴ࡟ࡼࡿ᪂ወ࢔࣏ࣝ࢟ࣝࣜࢢࣜࢥࢩࢻࡢ⏕ᡂ࡜≀ᛶゎᯒࢆ⾜ࡗࡓࠋᚤ⏕≀࡟࠾ࡅࡿ㛗㙐࢔ࣝࢥ࣮ࣝ௦ㅰ⤒㊰ࡣ NAD(P)+ࢆ㟁Ꮚཷᐜయ࡜ࡍࡿ࢔ࣝࢥ࣮ࣝࢹࣄࢻࣟࢤࢼ࣮ࢮ⣔ࠊࡶࡋࡃࡣ㓟⣲ࢆཷᐜయ࡜ࡍࡿ⬡⫫᪘࢔ࣝࢥ࣮ࣝ࢜࢟ࢩࢲ࣮ࢮ㸦fatty
alcohol oxidase㸸FAO㸧⤒㊰ࡀ▱ࡽࢀ࡚࠸ࡿࠋS. bombicola ࡢ 1 ⣭࢔ࣝࢥ࣮ࣝ㈨໬⤒㊰࡜ࡋ࡚ࠊ࣌ࣜࣉࣛࢬ࣒࡟ FAO άᛶࢆ᭷ࡍࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࡀࠊ㓄ิ᝟ሗࡸ㓝⣲ࡢ≉ᛶゎᯒࡣሗ࿌ࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛ࡲࡎࠊS. bombicola ࡢࢤࣀ࣒ゎᯒࢆ⾜ࡗࡓࠋ᪤▱ࡢ FAO
࡛࠶ࡿ Candida tropicalis FAO2 ࡜ప࠸┦ྠᛶ㸦35%㸧ࢆ♧ࡍ᪂つ࡞ FAO1 㑇ఏᏊࢆぢฟࡋࡓࠋ኱⭠⳦ࢆ⏝࠸࡚ S. bombicola ࡢ FAO1 㑇ఏᏊࢆ␗✀Ⓨ⌧ࡋࠊᶵ⬟ゎᯒࢆ⾜ࡗࡓ࡜ࡇࢁࠊ㙐㛗 4 ࠿ࡽ 16 ࡲ࡛ࡢ 1 ⣭࢔ࣝࢥ࣮ࣝ࡟ᑐࡋ㓟໬άᛶࢆ♧ࡋࠊFAO άᛶࢆ᭷ࡍࡿࡇ࡜
ࡀ᫂ࡽ࠿࡜࡞ࡗࡓࠋḟ࡟ S. bombicola ࡟࠾ࡅࡿᰤ㣴せồᛶᰴࡢ㑅ᢤ࠾ࡼࡧᙧ㉁㌿᥮⣔ࢆᵓ⠏ࡋࠊ┦ྠ⤌᥮࠼࡟ࡼࡿ FAO1 㑇ఏᏊࡢ◚ቯࢆヨࡳࡓࠋࡑࡢ⤖ᯝࠊFAO1 㑇ఏᏊḞኻᰴ㸦ΔFAO1㸧ࡣࠊ1 ⣭࢔ࣝࢥ࣮ࣝ࡟ᑐࡍࡿ㈨໬⬟࠾ࡼࡧቑṪ⬟ࢆኻ࠺ࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗࡓࠋ
ࡇࢀࡽࡢ⤖ᯝ࠿ࡽࠊS. bombicola ࡟࠾ࡅࡿ 1 ⣭࢔ࣝࢥ࣮ࣝࡢ୺ࡓࡿ௦ㅰ⤒㊰ࡣࠊFAO1 ࡟ࡼࡿ㓟໬࡛࠶ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋࡉࡽ࡟ࠊΔFAO1 ᰴࢆ⏝࠸࡚ࢢࣝࢥ࣮ࢫ࡜ 1-ࢸࢺࣛࢹ࢝ࣀ࣮ࣝࢆ཯ᛂࡉࡏࡓ⤖ᯝࠊ1-ࢸࢺࣛࢹ࢝ࣀ࣮ࣝ㦵᱁࠿ࡽᡂࡿࢯ࣍ࣟࣜࣆࢵࢻ⏕⏘⬟ࡢྥୖ࡜ࠊᮍ▱໬ྜ≀ࡀᚓࡽࢀࡓࠋヱ໬ྜ≀ࢆ⢭〇ᚋࠊLC-MS ཬࡧຍỈศゎᚋࡢ GC-MS ࡟ࡼࡗ࡚࢔ࣝ࢟ࣝᇶࡢᵓ㐀ࢆゎᯒࡋࡓ࡜ࡇࢁࠊᮍ▱໬ྜ≀ࢆᵓᡂࡍࡿ࢔ࣝ࢟ࣝᇶࡣ 1,13-ࢸࢺࣛࢹ࢝ࣥࢪ࣮࢜ࣝࡶࡋࡃࡣ 1,14-ࢸࢺࣛࢹ࢝ࣥࢪ࣮࡛࢜ࣝ࠶ࡾࠊࡇࢀࡽࢪ࣮࢜ࣝ࡟⢾㙐ࡀ 4ࡘ௜ຍࡋࡓ᪂つ࡞⢾⬡㉁࡛࠶ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋ୧ᮎ➃ࢪ࣮࢜ࣝࡢ㓄⢾య໬≀ࡣࡇࢀࡲ࡛ሗ࿌ࡀ࡯࡜ࢇ࡝ㄆࡵࡽࢀࡎࠊ᪂つᛶࡀ㧗࠸࡜⪃࠼ࡽࢀࡓࠋ ➨ 2 ❶࡛ࡣࠊ2 ⣭࢔ࣝࢥ࣮ࣝࡢ௦ㅰ㈨໬⤒㊰ࡢゎ᫂ࢆ⾜ࡗࡓࠋΔFAO1 ᰴࡣ 2 ⣭࢔ࣝࢥ࣮ࣝࢆ㈨໬࡛ࡁ࡞࠸ࡓࡵࠊFAO1 ࡣ 2 ⣭࢔ࣝࢥ࣮ࣝ௦ㅰ⤒㊰࡟࠾࠸࡚ࡶ୺ࡓࡿ㓟໬཯ᛂ࡜ࡋ
࡚ᐤ୚ࡍࡿ࡜⪃࠼ࡽࢀࡿࠋࡋ࠿ࡋ࡞ࡀࡽ FAO1 ࡣ 2 ⣭࢔ࣝࢥ࣮ࣝࢆᇶ㉁࡜ࡋ࡚࡯࡜ࢇ࡝฼
⏝ࡋ࡞࠸ࡓࡵࠊ࢔ࣝࢥ࣮ࣝศゎ࡟࠾ࡅࡿึⓎ㓟໬ẁ㝵ࡣࠊ2 ⣭࢔ࣝࢥ࣮ࣝᇶࡢ㓟໬࡛ࡣ࡞࠸࡜⪃࠼ࡽࢀࡓࠋࡑࡇ࡛ࠊΔFAO1 ᰴ࡟࠾࠸࡚ 2 ⣭࢔ࣝࢥ࣮ࣝ࠾ࡼࡧࢢࣝࢥ࣮ࢫࢆᇶ㉁࡜ࡋ
࡚ᇵ㣴ࡋࡓ࡜ࡇࢁࠊω ᮎ➃ࠊளᮎ➃ࡀỈ㓟໬ࡉࢀࡓࢪ࣮࢜ࣝᆺࡢ⢾⬡㉁ࡀᚓࡽࢀࡓࠋࡲࡓࠊ
ࢯ࣍ࣟࣜࣆࢵࢻྜᡂ࡟ᐤ୚ࡍࡿ⢾㌿⛣㓝⣲࡛࠶ࡿ UGTA1 㑇ఏᏊࢆḞᦆࡉࡏࡿ࡜ࠊୖグ⢾⬡㉁ࡣᾘኻࡋࠊω ᮎ➃ࠊளᮎ➃ࢪ࣮࢜ࣝࡢ⵳✚ࡀㄆࡵࡽࢀࡓࠋࡇࢀࡽࡢ⤖ᯝ࠿ࡽࠊ S.

bombicola ࡟࠾࠸࡚ࠊ2 ⣭࢔ࣝࢥ࣮ࣝࡣ ω ᮎ➃ࡢỈ㓟໬ࢆཷࡅࡓᚋࠊFAO1 ࡟ࡼࡾ㓟໬ࡉࢀࠊβ 㓟໬⤒㊰ࢆ฼⏝ࡋ࡚㈨໬ࡉࢀࡿ࡜⪃࠼ࡽࢀࡓࠋ௨ୖࡢ⤖ᯝ࠿ࡽࠊ⢾⬡㉁⏕⏘⳦࡛࠶ࡿ S. bombicola ࡟࠾࠸࡚ 1 ⣭࢔ࣝࢥ࣮ࣝ㓟໬㓝⣲
FAO1 ࢆྠᐃࡋࡓࠋFAO1 ࡣᮏ⳦࡟࠾࠸࡚㛗㙐࢔ࣝࢥ࣮ࣝ௦ㅰ⤒㊰࡟኱ࡁ࡞ᐤ୚ࢆ♧ࡍࡇ࡜ࢆド᫂ࡋࠊFAO1 㑇ఏᏊ◚ቯᰴ࡛ࡣᢞධࡉࢀࡓ࢔ࣝࢥ࣮ࣝࢆຠ⋡ࡼࡃ㓄⢾య࡜ࡋ࡚ኚ᥮࡛ࡁࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋࡇࢀࡽࡢ◊✲ᡂᯝࡣࠊS. bombicola ࠾ࡅࡿ࢔ࣝࢥ࣮ࣝ௦ㅰ⤒㊰ࡢゎ᫂࡟㔜せ࡞▱ぢࢆࡶࡓࡽࡍࡔࡅ࡛ࡣ࡞ࡃࠊ᪂ࡓ࡞㓄⢾యࠊ⢾⬡㉁ࠊ୧ᮎ➃ࢪ࣮࢜ࣝࡢྜᡂἲࢆ♧ࡍ㔜せ࡞◊✲࡜ࡋ࡚㧗ࡃホ౯ࡉࢀࡿ ௨ୖࠊᑂᰝࡢ⤖ᯝࠊᮏㄽᩥࡢⴭ⪅ࡣ༤ኈ㸦⌮Ꮫ㸧ࡢᏛ఩ࢆᤵ୚ࡉࢀࡿ༑ศ࡞㈨᱁ࡀ࠶ࡿ
ࡶࡢ࡜ㄆࡵࡿࠋබ⾲ㄽᩥ
Identification of the fatty alcohol oxidase FAO1 from Starmerella bombicola and improved novelglycolipids production in an FAO1 knockout mutant.
Fumikazu Takahashi, Kazuaki Igarashi and Hiroshi Hagihara.
Applied Microbiology and Biotechnology 100: 9519-9528 (2016)
ཧ⪃ㄽᩥ
Increased dipicolinic acid production with an enhanced spoVF operon in Bacillus subtilis andmedium optimization.
Fumizazu Takahashi, Nobuyuki Sumitomo, Hiroshi Hagihara and Katsuya Ozaki.
Biotechnology and Biochemistry 79: 505-511 (2015)
Characterization of a novel glucosamine-6-phosphate deaminase from a hyperthermophilicarchaeon.
Takeshi Tanaka, Fumikazu Takahashi, Toshiaki Fukui, Shinsuke Fujiwara, Haruyuki Atomi andTadayuki Imanaka.
Journal of Bacteriology 187: 7038-7044 (2005)