Molecular basis for oligosaccharide epimerases and their related enzymes.

• Project/Area Number: 26850059
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: SABURI Wataru 北海道大学, (連合)農学研究科(研究院), 助教 (00598089)
• 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: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: セロビオース2-エピメラーゼ / エピメラーゼ / イソメラーゼ / オリゴ糖 / 部位特異的変異 / 基質特異性 / 反応特異性

Outline of Final Research Achievements

Cellobiose 2-epimerase (CE) epimerizes the reducing end glucose residue of beta1-4 disaccharides to mannose residue. It shares the catalytic domain and site structures with other monosaccharide isomerases/epimerases. In this study, structure-function relationship of these enzymes was analyzed. Important amino acid residues for high selectivity for disaccharides in Rhodothermus marines CE were determined through site-directed mutation. Slight isomerization activity was observed in R. marines CE, and important structure for isomerase activity was determined based on the comparison of the structures between R. marines and Cardicellulosiruptor saccharolyticus CEs. A novel enzyme acting on mannose was found from function unknown proteins.

Research Products

• [Journal Article] Functions, structures, and applications of cellobiose 2-epimerase and glycoside hydrolase family 130 mannoside phosphorylases. (2016)
  • Author(s): Saburi W.
  • Journal Title: Biosci Biotechnol Biochem. Volume: 80 Issue: 7 Pages: 1294-1305
  • DOI: 10.1080/09168451.2016.1166934
  • NAID: 120006360137
  • Peer Reviewed / Acknowledgement Compliant
• [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] Supplemental epilactose prevents metabolic disorders through uncoupling protein-1 induction in the skeletal muscle of mice fed high-fat diets. (2015)
  • Author(s): Murakami Y, Ojima-Kato T, Saburi W, Mori H, Matsui H, Tanabe S, Suzuki T
  • Journal Title: Br. J. Nutr. Volume: 114 Issue: 11 Pages: 1774-1783
  • DOI: 10.1017/s0007114515003505
  • Peer Reviewed
• [Journal Article] Biochemical properties and substrate recognition mechanism of GH31 α-glucosidase from Bacillus sp. AHU 2001 with broad substrate specificity (2015)
  • Author(s): Wataru Saburi, Masayuki Okuyama, Yuya Kumagai, Atsuo Kimura, Haruhide Mori
  • Journal Title: Biochimie Volume: 108 Pages: 140-148
  • DOI: 10.1016/j.biochi.2014.11.010
  • Peer Reviewed
• [Journal Article] Structural insights into catalytic reaction that is involved in the reorientation of Trp238 at the substrate-binding site in GH13 dextran glucosidase (2015)
  • Author(s): Kobayashi M, Saburi W, Nakatsuka D, Hondoh H, Kato K, Okuyama M, Mori H, Kimura A, and Yao M
  • Journal Title: FEBS Letters Volume: 589 Issue: 4 Pages: 484-489
  • DOI: 10.1016/j.febslet.2015.01.005
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural elements responsible for the glucosidic linkage-selectivity of a glycoside hydrolase family 13 exo-glucosidase (2015)
  • Author(s): Wataru Saburi, Hiroaki Rachi-Otsuka, Hironori Hondoh, Masayuki Okuyama, Haruhide Mori, Atsuo Kimura
  • Journal Title: FEBS letters Volume: 589 Issue: 7 Pages: 865-869
  • DOI: 10.1016/j.febslet.2015.02.023
  • NAID: 120005725141
  • Peer Reviewed
• [Journal Article] Colorimetric Quantification of β-(1→4)-Mannobiose and 4-<i>O</i>-β-D-Mannosyl-D-glucose (2014)
  • Author(s): Nongluck Jaito, Wataru Saburi, Hirohiko Muto, Hirokazu Matsui, Haruhide Mori
  • Journal Title: Journal of Applied Glycoscience Volume: 61 Issue: 4 Pages: 117-119
  • DOI: 10.5458/jag.jag.JAG-2014_007
  • NAID: 130004699355
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed / Acknowledgement Compliant
• [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
• [Journal Article] Acidophilic β-Galactosidase from <i>Aspergillus niger</i> AHU7120 with Lactose Hydrolytic Activity Under Simulated Gastric Conditions (2014)
  • Author(s): Wataru Saburi, Hiroshi M Ueno, Hirokazu Matsui, Haruhide Mori
  • Journal Title: Journal of Applied Glycoscience Volume: 61 Issue: 2 Pages: 53-57
  • DOI: 10.5458/jag.jag.JAG-2013_011
  • NAID: 130004481038
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray crystallographic analysis of alpha-glucosidase HaG from Halomonas sp. strain H11 (2014)
  • Author(s): X. Shen, W. Saburi, Z.-Q. Gai, K. Komoda, J. Yu, T. Ojima-Kato, Y. Kido, H. Matsui, H. Mori and M. Yao
  • Journal Title: Acta Crystallographica Section F Volume: F70 Issue: 4 Pages: 464-466
  • DOI: 10.1107/s2053230x14001940
  • Peer Reviewed
• [Journal Article] Enhancement of hydrolytic activity of thermophilic alkalophilic α-amylase from Bacillus sp. AAH-31 through optimization of amino acid residues surrunding the substrate binding site. (2014)
  • Author(s): Naoya Tamamura, Wataru Saburi, Atsushi Mukai, Naoki Morimoto, Toshihiko Takehana, Seiji Koike, Hirokazu Matsui, Haruhide Mori
  • Journal Title: Biochemical Engineering Journal Volume: 86 Pages: 8-15
  • DOI: 10.1016/j.bej.2014.02.014
  • Peer Reviewed
• [Journal Article] Identity of the two dextran dextrinases produced by Gluconobacter oxydans ATCC 11894 and its localization change depending on the cell growth (2014)
  • Author(s): Juri Sadahiro, Haruhide Mori, Wataru Saburi, Masayuki Okuyama, Atsuo Kimura
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 456 Issue: 1 Pages: 500-505
  • DOI: 10.1016/j.bbrc.2014.11.115
  • Peer Reviewed
• [Presentation] 乳中ラクトースのエピラクトースへの直接変換を志向した各種セロビオース2-エピメラーゼの機能評価 (2016)
  • Author(s): 武藤 洋彦，佐分利 亘，森 春英
  • Organizer: 日本農芸化学会2016年度大会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-03-30
• [Presentation] Ruminococcus albus由来4-O-β-D-マンノシル-D-グルコースホスホリラーゼのサブサイト+1に保存されるArg92の基質結合への関与 (2016)
  • Author(s): 塩田 咲耶子，尾高 伶，佐分利 亘，イエ ウキン，薦田 圭介，加藤 公児，西本 完，北岡 本光，姚 閔，森 春英
  • Organizer: 日本農芸化学会2016年度大会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-03-30
• [Presentation] 嫌気性細菌によるエピラクトース代謝の多様性 (2016)
  • Author(s): 坂井 未悠，佐分利 亘，森 春英
  • Organizer: 日本農芸化学会2016年度大会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-03-28
• [Presentation] オリゴ糖異性化酵素および関連酵素の分子基盤と応用 (2015)
  • Author(s): 佐分利 亘
  • Organizer: 平成27年度 公益社団法人 日本農芸化学会 北海道支部第2回講演会
  • Place of Presentation: 北海道大学（北海道札幌市）
  • Year and Date: 2015-11-14
  • Invited
• [Presentation] Ruminococcus albus由来4-O-β-D-mannosyl-D-glucose phosphorylase RaMP1の無機リン酸に対するホモトロピック相互作用 (2015)
  • Author(s): 尾髙 伶，佐分利 亘，Ye Yuxin，薦田 圭介，加藤 公児，西本 完，北岡 本光，姚 閔，森 春英
  • Organizer: 日本応用糖質科学会平成27年度大会
  • Place of Presentation: 奈良春日野国際フォーラム（奈良県奈良市）
  • Year and Date: 2015-09-16
• [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 Bioenginnering Meeting
  • Place of Presentation: Dipoli congress centre (Espoo, Finland)
  • Year and Date: 2015-05-10
  • Int'l Joint Research
• [Presentation] Biochemical characterization of a novel aldose-ketose isomerase, mannose isomerase from Marinomonas mediterranea (2015)
  • Author(s): Nongluck Jaito, Wataru Saburi, Yuka Tanaka, and Haruhide Mori
  • Organizer: 11th Carbohydrate Bioenginnering Meeting
  • Place of Presentation: Dipoli congress centre (Espoo, Finland)
  • Year and Date: 2015-05-10
  • Int'l Joint Research
• [Presentation] Bacillus sp. AHU 2001由来GH31 α-グルコシダーゼの生化学的諸性質と基質認識に重要な構造因子の解析 (2015)
  • Author(s): 佐分利 亘，奥山 正幸，熊谷 祐也，木村 淳夫，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: 2015-03-28
• [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] 低温に応答したGA量の抑制によりイネの耐性を高めることができる (2014)
  • Author(s): 高橋 直希，川又 彩，手塚 大介，佐分利 亘，松浦 英幸，森 春英，今井 亮三
  • Organizer: 植物化学調節学会第49回大会
  • Place of Presentation: 京都大学 (京都市)
  • Year and Date: 2014-10-18 – 2014-10-19
• [Presentation] 傷害時のイネにおけるサリチル酸グルコシドを糖ドナーとするツベロン酸への糖転移反応 (2014)
  • Author(s): 竹松 知紀，瀬戸 義哉，宮澤 吉郎，和久田 真司，佐分利 亘，森 春英，高橋 公咲，松浦 英幸
  • Organizer: 植物化学調節学会第49回大会
  • Place of Presentation: 京都大学 (京都市)
  • Year and Date: 2014-10-18 – 2014-10-19
• [Presentation] イネのトレハロースにより誘導されるシステミックな病害抵抗性にはジャスモン酸が関与する (2014)
  • Author(s): 手塚 大介，坂井 志帆，和久田 真司，加藤 英樹，松浦 英幸，佐分利 亘，森 春英，松井 博和，今井 亮三
  • Organizer: 植物化学調節学会第49回大会
  • Place of Presentation: 京都大学 (京都市)
  • Year and Date: 2014-10-18 – 2014-10-19
• [Presentation] イネの傷害応答におけるサリチル酸グルコシド加水分解酵素の機能 (2014)
  • Author(s): 武田 遼介，佐分利 亘，姫野 奈美，和久田 真司，松浦 英幸，今井 亮三，松井 博和，森 春英
  • Organizer: 植物化学調節学会第49回大会
  • Place of Presentation: 京都大学 (京都市)
  • Year and Date: 2014-10-18 – 2014-10-19
• [Presentation] イネ由来スクロースシンターゼ3のヌクレオチド二リン酸特異性に関わるアミノ酸残基の決定 (2014)
  • Author(s): 岩藤 伸治，佐分利 亘，松井 博和，今井 亮三，森 春英
  • Organizer: 日本応用糖質科学会平成26年度大会
  • Place of Presentation: 朱鷺メッセ新潟コンベンションセンター (新潟市)
  • Year and Date: 2014-09-24
• [Presentation] Halonomans sp. H11株由来α-グルコシダーゼの糖転移機構の解析 (2014)
  • Author(s): 城戸 悠輔，佐分利 亘，小島 晃代，Xing Shen，薦田 圭介，姚 閔，松井 博和，森 春英
  • Organizer: 日本農芸化学会 北海道・東北合同支部大会
  • Place of Presentation: 北海道大学 (札幌市)
  • Year and Date: 2014-09-23
• [Remarks] 北海道大学大学院農学研究院生物化学研究室
  • URL: http://www.agr.hokudai.ac.jp/biochem/
• [Remarks] 北海道大学大学院農学研究院 生物化学研究室
  • URL: http://www.agr.hokudai.ac.jp/biochem/