Development of a highly sensitive method for stable isotope analysis of amino acids

• Project/Area Number: 17H02988
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Geochemistry/Cosmochemistry
• Research Institution: Hokkaido University
• Principal Investigator: Chikaraishi Yoshito 北海道大学, 低温科学研究所, 教授 (50455490)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2019: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000)
• Keywords: アミノ酸 / 安定同位体比 / GC-IRMS / 高感度 / 食物連鎖 / エネルギー / 起源 / 物質循環 / 安定同位体 / 地球化学 / 生態学 / 同位体比安定

Outline of Final Research Achievements

In this study, I developed a highly sensitive method for stable isotope analysis of amino acids, based on the incorporation of the following 4 techniques: (1) redesign of unions for connecting capillary column with reactors and traps; (2) increase of reactivity for reactors with metal catalyst; (3) installation of a multiple purpose sampler (MPS); and utilization of column coupling method. The results indicate that GC-IRMS modified in this study has a high sensitivity by approximately 100 times compared to the commercial GC-IRMS.

Academic Significance and Societal Importance of the Research Achievements

これまでアミノ酸の安定同位体比を用いた研究は，測定に必要な試料量が多いことが，重大な欠点であった。しかし，本研究により，上記(1)～(4)の技術を導入し，実際に，約100倍の感度の向上が行えたことで，本手法の応用範囲は劇的に広がったと考えている。試料サイズの制約の改善は，生物・生態系とその生合成・代謝系の機能や進化，地球生物圏における有機物・エネルギー循環，宇宙空間や初期地球における有機物生成や生命の起源に関連した研究など，幅広い分野への波及効果が，期待できる。

Research Products

• [Int'l Joint Research] ウィスコンシン大学(米国)
• [Int'l Joint Research] ウイスコンシン大学/ミシガン州立大学/ハワイ大学(米国)
• [Int'l Joint Research] ハンヤン大学(韓国)
• [Int'l Joint Research] Wisconsin University/Michigan Starte University(米国)
• [Journal Article] Quantifying niche partitioning and multichannel feeding among tree squirrels (2019)
  • Author(s): Pauli j.N., Manlick P.J., Dharampal P.S., Takizawa Y., Chikaraishi Y., Niccolai L.J., Grauer J.A., Black K.L., Restrepo G.M., Perring P.L., Wilson E.C., Martin M.E., Curras M.R., Bougie T.A., Thomson K.L., Smith M.M., Steffan S.A
  • Journal Title: Food Webs Volume: 21 Pages: e00124-e00124
  • DOI: 10.1016/j.fooweb.2019.e00124
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Omnivory in Bees: Elevated Trophic Positions among All Major Bee Families (2019)
  • Author(s): Steffan S.A., Dharampal P.S., Danforth B.N., Gaines-Day H.R., Takizawa Y., Chikaraishi Y.
  • Journal Title: The American Naturalist Volume: 194 Issue: 3 Pages: 414-421
  • DOI: 10.1086/704281
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular and isotopic compositions of nitrogen-containing organic molecules formed during UV-irradiation of simulated interstellar ice (2019)
  • Author(s): Sugahara H. , Takano Y. , Tachibana S. , Sugawara I. , Y. Chikaraishi , Ogawa N. O. , Ohkouchi N., Kouchi A. and Yurimoto H.
  • Journal Title: GEOCHEMICAL JOURNAL Volume: 53 Issue: 1 Pages: 5-20
  • DOI: 10.2343/geochemj.2.0553
  • NAID: 130007594396
  • ISSN: 0016-7002, 1880-5973
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insight into anaerobic methanotrophy from 13C/12C-amino acids and 14C/12C-ANME cells in seafloor microbial ecology (2018)
  • Author(s): Takano Yoshinori、Chikaraishi Yoshito、Imachi Hiroyuki、Miyairi Yosuke、Ogawa Nanako O.、Kaneko Masanori、Yokoyama Yusuke、Kr?ger Martin、Ohkouchi Naohiko
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 14070-14070
  • DOI: 10.1038/s41598-018-31004-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Compound-specific isotope analysis of benthic foraminifer amino acids suggests microhabitat variability in rocky-shore environments. (2018)
  • Author(s): Tsuchiya M, Chikaraishi Y, Nomaki H, Sasaki Y, Tame A, Uematsu K, Ohkouchi N
  • Journal Title: Ecology and Evolution Volume: 00 Issue: 16 Pages: 1-16
  • DOI: 10.1002/ece3.4358
  • Peer Reviewed / Open Access
• [Presentation] Trophic position estimate in a marine ecosystem with a wide δ15N baseline (2019)
  • Author(s): Xing Daochao, Chikaraishi Yoshito
  • Organizer: International Symposium on Isotope Physiology, Ecology, and Geochemistry 2019
  • Int'l Joint Research
• [Presentation] Trophic isotopic discrimination (2019)
  • Author(s): Chikaraishi Yoshito
  • Organizer: International Symposium on Isotope Physiology, Ecology, and Geochemistry 2019
  • Int'l Joint Research / Invited
• [Presentation] Remarkable fractionation of nitrogen isotopes in anaerobic amino acid metabolisms. (2019)
  • Author(s): Takizawa Yuko, Chikaraishi Yoshito
  • Organizer: The 29th International Meetings on Organic Geochemistry
  • Int'l Joint Research
• [Presentation] 生態系のエネルギー循環からみた共生系 (2019)
  • Author(s): 力石嘉人
  • Organizer: 共生起源研究会
  • Invited
• [Presentation] Habitat use of organisms viewed with an Isoscape map, based on nitrogen isotopic composition of amino acids (2018)
  • Author(s): Yoshito Chikaraishi
  • Organizer: International workshop on new application of stable isotope approaches
  • Int'l Joint Research / Invited
• [Presentation] Compound-specific isotope analysis (CSIA) of 15N/14N in amino acids: as a potential powerful tool for studying physiology, ecology, and geochemistry (2018)
  • Author(s): Yoshito Chikaraishi
  • Organizer: 42nd International Symposium on Capillary Chromatograph and 15th GCxGC Symposium
  • Int'l Joint Research / Invited
• [Presentation] 食物連鎖の中に存在する嫌気的代謝プロセスの可視化：蜜蜂が姿を消した原因を探る (2018)
  • Author(s): 力石嘉人，滝沢侑子，Steffan, A.S.
  • Organizer: 有機地球化学会品川シンポジウム
• [Presentation] Is Isotopic Fractionation in Carbon Isotopes Coupling with That in Nitrogen Isotopes in Food Webs? (2018)
  • Author(s): Chikaraishi Y., Takizawa Y.
  • Organizer: AOGS2018
  • Int'l Joint Research
• [Presentation] アミノ酸の窒素同位体比を用いた 「生物の栄養段階」推定法 (2018)
  • Author(s): 力石嘉人
  • Organizer: 日本生態学学会
  • Invited
• [Presentation] Stable isotope analysis of amino acids: as a new tool for illustrating central biosynthetic and metabolic flux. (2017)
  • Author(s): Chikaraishi Y., Takizawa Y.
  • Organizer: The 28th International Meeting of Organic Geochemistry
  • Int'l Joint Research
• [Remarks] 機化合物の安定同位体比の持つポテンシャル | GC-IRMS
  • URL: http://www.lowtem.hokudai.ac.jp/isophysiol/products/cn6/pg233.html
• [Funded Workshop] International Symposium on Isotope Physiology, Ecology, and geochemistry (2019)