Development of new beta-glucosidase activated with its catalytic product

• Project/Area Number: 26660081
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: MORI HARUHIDE 北海道大学, (連合)農学研究科(研究院), 教授 (80241363)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: β-グルコシダーゼ / バイオマス糖化 / 酵素活性化 / 反応機構 / セルロース / 酵素糖化 / 生成物阻害 / バイオマス / 糖化

Outline of Final Research Achievements

Enhancement of activity of beta-glucosidase from Streptomyces sp. with the product glucose was investigated.. Kinetics analyses with various substrates and monosaccharides revealed that the activation is caused not only glucose but also other monosaccharides as well as even short-chain substrates with solely drastic increase in kcat. The monosaccharides, partially occupying the far side of substrate binding site from the catalytic site, prevent the short-chain substrates from binding in the non-productive fashion. Based on possible amino acid residues associated to the monosaccharide binding, candidate beta-glucosidases equipped with the activation feature can be selected.

Research Products

• [Journal Article] Structural insights into the difference in substrate recognition of two mannoside phosphorylases from two GH130 subfamilies (2016)
  • Author(s): Ye, Y., Saburi, W., Odaka, R., Kato, K., Sakurai, N., Komoda, K., Nishimoto, M., Kitaoka, M., Mori, H., and Yao, M.
  • Journal Title: FEBS Lett Volume: 590 Issue: 6 Pages: 828-837
  • DOI: 10.1002/1873-3468.12105
  • Peer Reviewed
• [Journal Article] A single-nucleotide polymorphism in an endo-β-glucanase gene controles seed coat permeability in soybean (2015)
  • Author(s): Jang, S.J., Sato, M., Sato, K., Jitsuyama, Y., Fujino, K., Mori, H., Takahashi, R., Benitez, E.R., Liu, B., Yamada, T., and Abe, J.
  • Journal Title: PLoS One Volume: 10 Issue: 6 Pages: e0128527-e0128527
  • DOI: 10.1371/journal.pone.0128527
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Identification of rice Os4BGlu13 as a β-glucosidase which hydrolyzes gibberellin A4 1-O-β-D-glucosyl ester, in addition to tuberonic acid glucoside and salicylic acid derivative glucosides. (2015)
  • Author(s): Hua, Y., Ekkhara, W., Sansenya, S., Srisomsap, C., Roytakul, S., Saburi, W., Takeda, R., Matsuura, H., Mori, H., and Ketudat Cairns, J.R.
  • Journal Title: Arch. Biochem. Biophys. Volume: 583 Pages: 36-46
  • DOI: 10.1016/j.abb.2015.07.021
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structural insights into the epimerization of β-1,4-linked oligosaccharides catalyzed by cellobiose 2-epimerase, the sole enzyme epimerizaing non-anomeric hydroxyl groups of unmodified sugars (2014)
  • Author(s): Takaaki Fujiwara, Wataru Saburi, Hirokazu Matsui, Haruhide Mori, Min Yao
  • Journal Title: Journal of Biological Chemistry Volume: 289 Issue: 6 Pages: 3405-3415
  • DOI: 10.1074/jbc.m113.531251
  • Peer Reviewed
• [Journal Article] Characterization of a thermophilic 4-O-β-D-mannosyl-D-glucose phosphorylase from Rhodothermus marinus. (2014)
  • Author(s): Nongluck Jaito, Wataru Saburi, Rei Odaka, Yusuke Kido, Ken Hamura, Mamoru Nishimoto, Motomitsu Kitaoka, Hirokazu Matsui, Haruhide Mori
  • Journal Title: Bioscience Biotechnology Biochemistry Volume: 78 Issue: 2 Pages: 263-270
  • DOI: 10.1080/09168451.2014.882760
  • Peer Reviewed
• [Presentation] Molecular basis for the epimerization of oligosaccharides by cellobiose 2-epimerase (2015)
  • Author(s): Wataru Saburi, Takaaki Fujiwara, Nongluck Jaito, Hirohiko Muto, Hirokazu Matsui, Min Yao, and Haruhide Mori
  • Organizer: 11th Carbohydrate Bioengineering Meeting
  • Place of Presentation: Espoo, Finland
  • Year and Date: 2015-05-10
  • Int'l Joint Research
• [Presentation] Characterization of the recombinant cellobiose 2-epimerase from Rhodothermus marinus and site-directed mutagenesis of the catalytic residues. (2015)
  • Author(s): Nongluck Jaito, Wataru Saburi, Hirohiko Muto, Takaaki Fujiwara, Min Yao, and Haruhide Mori
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学（岡山市）
  • Year and Date: 2015-03-27
• [Presentation] Rhodothermus marinus JCM9785由来セロビオース2-エピメラーゼの基質結合部位周辺アミノ酸残基の機能解析 (2015)
  • Author(s): 武藤 洋彦，佐分利 亘，藤原 孝彰，姚 閔，森 春英
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学（岡山市）
  • Year and Date: 2015-03-27
• [Presentation] グライコシンターゼ反応を利用したシロイヌナズナ由来β-グルコシダーゼBglu15のアグリコン特異性の解析 (2015)
  • Author(s): 菅原 好美，佐分利 亘，谷口 沙希，今井 亮三，森 春英
  • Organizer: 日本農芸化学会2015年度大会
  • Place of Presentation: 岡山大学（岡山市）
  • Year and Date: 2015-03-27
• [Presentation] GH130 β-マンノシドホスホリラーゼ：発見､生理的機能､酵素反応 (2014)
  • Author(s): 森春英、佐分利亘、松井博和
  • Organizer: 日本糖質学会年会
  • Place of Presentation: 名古屋大学（名古屋市）
  • Year and Date: 2014-08-11