Development of novel assay of beta-glycanases for complete enzymatic saccharification of unused biomass

• Project/Area Number: 24380089
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Wood science
• Research Institution: The University of Tokyo
• Principal Investigator: IGARASHI Kiyohiko 東京大学, 農学生命科学研究科, 准教授 (80345181)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000); Fiscal Year 2014: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2012: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
• Keywords: バイオマス / βーグリカナーゼ / セルラーゼ / キチナーゼ / 結晶性多糖 / セルロース / キチン / プロセッシビティ / 糖化 / グリカナーゼ

Outline of Final Research Achievements

Cellulose and chitin are the main components of unused biomass, and effective utilization of these materials is expected as source of carbon of the next generation without competition with food. Considering the polysaccharides share the common feature consisting with beta-linked glycosides, it is the key to utilize the unused biomass how to control degrading enzymes efficiently at a solid/liquid interface.
In the present research, the behavior of beta-glycanase such as cellulase from filamentous fungi and cellulase and chitinase from bacteria was observed qualitatively and quantitatively at a solid/liquid interface by high-speed atomic force microscopy, and a strategy for the effective hydrolysis of unused biomass using a biochemical process was built.

Research Products

• [Journal Article] Trade-off between Processivity and Hydrolytic Velocity of Cellobiohydrolases at the Surface of Crystalline Cellulose (2014)
  • Author(s): Akihiko Nakamura, Watanabe, H., Ishida, T., Uchihashi, T., Wada, M., Ando, T., Igarashi, K. and Samejima, M.
  • Journal Title: Journal of the American Chemical Society Volume: 136 Issue: 12 Pages: 4584-4592
  • DOI: 10.1021/ja4119994
  • Peer Reviewed
• [Journal Article] Single-molecule Imaging Analysis of Elementary Reaction Steps of Trichoderma reesei Cellobiohydrolase I (Cel7A) Hydrolyzing Crystalline Cellulose Iα and IIII (2014)
  • Author(s): Y.Shibafuji, A.Nakamura, T.Uchihashi, N.Sugimoto, S.Fukuda, H.Watanabe, M.Samejima, T.Ando, H.Noji, A.Koivula, K.Igarashi, R.Iino
  • Journal Title: Journal of Biological Chemistry Volume: 289 (29) Issue: 20 Pages: 1405614065-1405614065
  • DOI: 10.1074/jbc.m113.546085
  • Peer Reviewed / Open Access
• [Journal Article] Two-way traffic of glycoside hydrolase family 18 processive chitinases on crystalline chitin (2014)
  • Author(s): Igarashi, K., Uchihashi, T., Uchiyama, T., Sugimoto, H., Wada, M., Suzuki, K., Sakuda, S., Ando, T., Watanabe, T., and Samejima, M.
  • Journal Title: Nature Commun. Volume: 5 Issue: 1 Pages: 3975-3975
  • DOI: 10.1038/ncomms4975
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] High-speed atomic force microscope combined with single-molecule fluorescence microscope. (2013)
  • Author(s): S. Fukuda, T. Uchihashi, R. Iino, Y. Okazaki, M. Yoshida, K. Igarashi, T. Ando
  • Journal Title: Rev. Sci. Instrum. Volume: 84 Issue: 7 Pages: 73706-73706
  • DOI: 10.1063/1.4813280
  • Peer Reviewed
• [Journal Article] The Tryptophan Residue at the Active Site Tunnel Entrance of Trichoderma reesei Cellobiohydrolase Ce17A Is Important for Initiation of Degradation of Crystalline Cellulose (2013)
  • Author(s): Akihiko Nakamura, Tsukada, T., Auer, S., Furuta, T., Wada, M., Koivula, A., Igarashi, K. and Samejima M.
  • Journal Title: Journal of Biological Chemistry Volume: 288 Issue: 19 Pages: 13503-13510
  • DOI: 10.1074/jbc.m113.452623
  • Peer Reviewed
• [Journal Article] Degradation of Crystalline Celluloses by <I>Phanerochaete chrysosporium </I>Cellobiohydrolase II (Cel6A) Heterologously Expressed in Methylotrophic Yeast <I>Pichia pastoris</I> (2012)
  • Author(s): Igarashi, K., Maruyama, M., Nakamura, A., Ishida, T., Wada, M., and Samejima, M.
  • Journal Title: Journal of Applied Glycoscience Volume: 59 Issue: 3 Pages: 105-110
  • DOI: 10.5458/jag.jag.JAG-2011_029
  • NAID: 130004481005
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed
• [Journal Article] Adsorption characteristics of fungal family 1 cellulose-binding domain from Trichoderma reesei cellobiohydrolase I on crystalline cellulose: Negative cooperative adsorption via a steric exclusion effect (2012)
  • Author(s): Sugimoto, N., Igarashi, K., Wada, M., and Samejima, M.
  • Journal Title: Langmuir Volume: 28 Issue: 40 Pages: 14323-14329
  • DOI: 10.1021/la302352k
  • Peer Reviewed
• [Journal Article] Visualization of cellobiohydrolase I from Trichoderma reesei moving on crystalline cellulose using high-speed atomic force microscopy (2012)
  • Author(s): Kiyohiko Igarashi
  • Journal Title: Methods in Enzymology Volume: 510 Pages: 169-182
  • DOI: 10.1016/b978-0-12-415931-0.00009-4
  • Peer Reviewed
• [Presentation] セルロース資化性放線菌Cellulomonas fimi由来のセルラーゼの性状解析 (2015)
  • Author(s): 内山拓、中村彰彦、五十嵐圭日子、鮫島正浩、金子哲、内橋貴之
  • Organizer: 2015年度農芸化学会大会
  • Place of Presentation: 岡山大学津島キャンパス 岡山
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] Processivity in enzymatic degradation of crystalline polysaccharides: cellulases and chitinases (2014)
  • Author(s): Kiyohiko Igarashi
  • Organizer: Mie Bioforum 2014
  • Place of Presentation: Nemuno Sato Hotel & Resort 三重
  • Year and Date: 2014-11-18 – 2014-11-21
  • Invited
• [Presentation] セルロース系バイオマスを分解する酵素の構造機能相関 (2014)
  • Author(s): 五十嵐圭日子
  • Organizer: 学術振興会第169委員会 第45回研究会
  • Place of Presentation: ゆうぽうと 東京
  • Year and Date: 2014-07-25
  • Invited
• [Presentation] 糖質加水分解酵素ファミリー６セルラーゼのプロセッシビティを中心とした機能解析 (2014)
  • Author(s): 内山拓、中村彰彦、五十嵐圭日子、鮫島正浩、金子哲、内橋貴之
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 東京
• [Presentation] Cellulases: cooperative biomass breakdown (2014)
  • Author(s): Kiyohiko Igarashi
  • Organizer: JST-ALCA international workshop for plant cell wall engineering
  • Place of Presentation: Tokyo
  • Invited
• [Presentation] 結晶性セルロースとキチンを分解する酵素 : プロセッシブな加水分解の分子機構とドメイン構造における戦略 (2013)
  • Author(s): 五十嵐圭日子
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学(招待講演)
  • Year and Date: 2013-03-27
• [Presentation] 微生物におけるセルロースとキチンの酵素分解戦略：結晶性構造多糖を利用するためのヒント (2013)
  • Author(s): 五十嵐圭日子
  • Organizer: 第7回多糖の未来フォーラム
  • Place of Presentation: 大阪
  • Invited
• [Presentation] プロセッシブグリコシダーゼはどうやって結晶性多糖の上を走るのか？ (2013)
  • Author(s): 五十嵐圭日子
  • Organizer: 第3回分子モーター討論会
  • Place of Presentation: 東京
  • Invited
• [Book] 植物細胞壁 (2013)
  • Author(s): 五十嵐圭日子
  • Total Pages: 349
  • Publisher: 講談社
• [Remarks] やはりカメはウサギに勝る
  • URL: http://www.u-tokyo.ac.jp/ja/utokyo-research/research-news/the-tortoise-beats-the-hare-again/
• [Remarks] 分解酵素にとって結晶性キチンは双方向の高速道路だった
  • URL: http://www.a.u-tokyo.ac.jp/topics/2014/20140605-1.html
• [Remarks] 高速で逆送する酵素がキチンを分解
  • URL: http://scienceportal.jst.go.jp/news/newsflash_review/newsflash/2014/06/20140609_03.html
• [Remarks] 糸状菌由来セルラーゼがセルロース分子鎖の末端を認識する機構の解明
  • URL: http://www.a.u-tokyo.ac.jp/topics/2013/20130510-2.html
• [Remarks] 結晶性セルロースの分解は、切断の「速さ」ではなく基質を掴み続ける「ねばり強さ」が重要である
  • URL: http://www.a.u-tokyo.ac.jp/topics/2014/20140313-4.html
• [Remarks] やはりカメはウサギに勝る
  • URL: http://www.u-tokyo.ac.jp/ja/utokyo-research/research-news/the-tortoise-beats-the-hare-again/