Study on the role of uncharacterized enzymes belonging to glycoside hydrolase family 131 in biomass degradation

• Project/Area Number: 26660277
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental agriculture(including landscape science)
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: Tonozuka Takashi 東京農工大学, (連合)農学研究科(研究院), 准教授 (50285194)
• Co-Investigator(Renkei-kenkyūsha): YOSHIDA MAKOTO 東京農工大学, 大学院農学研究院, 准教授 (30447510)
• 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: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: バイオマス / 担子菌 / 糖質加水分解酵素

Outline of Final Research Achievements

This study focuses on enzymes belonging to glycoside hydrolase family 131, CcGH131A and CcGH131B, from a basidiomycete, Coprinopsis cinerea. A heterologous expression system of CcGH131A using Pichia pastoris was constructed, and the crystal structure of CcGH131A in a complex with ligand was determined. The cDNA of CcGH131B was cloned, and a heterologous expression system of CcGH131A using Escherichia coli was constructed. The protein was crystallized and the structure was determined. An experiment using fluorescence spectroscopy was performed to analyze the interaction between the proteins and polysaccharides. The results suggest that the substrate specificities of CcGH131A and CcGH131B are expected to be markedly different.

Research Products

• [Journal Article] A Surface Loop in the N-Terminal Domain of <i>Pedobacter heparinus </i>Heparin Lyase II is Important for Activity (2016)
  • Author(s): Mori, M., Ichikawa, M., Kiguchi, Y., Miyazaki, T., Hattori, M., Nishikawa, A. and Tonozuka, T.
  • Journal Title: Journal of Applied Glycoscience Volume: 63 Issue: 1 Pages: 7-11
  • DOI: 10.5458/jag.jag.JAG-2015_019
  • NAID: 130005126301
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed / Open Access
• [Journal Article] A glycoside hydrolase family 31 dextranase with high transglucosylation activity from Flavobacterium johnsoniae (2016)
  • Author(s): Gozu, Y., Ishizaki, Y., Hosoyama, Y., Miyazaki, T., Nishikawa, A. and Tonozuka, T.
  • Journal Title: Biosci. Biotechnol. Biochem. Volume: 印刷中 Issue: 8 Pages: 1562-1567
  • DOI: 10.1080/09168451.2016.1182852
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structural and biochemical characterization of novel bacterial α-galactosidases belonging to glycoside hydrolase family 31 (2015)
  • Author(s): Takatsugu Miyazaki, Yuichi Ishizaki, Megumi Ichikawa, Atsushi Nishikawa and Takashi Tonozuka
  • Journal Title: Biochemical Journal Volume: in press Issue: 1 Pages: 145-158
  • DOI: 10.1042/bj20150261
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Crystal structure and mutational analysis of isomalto-dextranase, a member of glycoside hydrolase family 27 (2015)
  • Author(s): Okazawa, Y., Miyazaki, T., Yokoi, G., Ishizaki, Y., Nishikawa, A. and Tonozuka, T.
  • Journal Title: J. Biol. Chem. Volume: 290 Issue: 43 Pages: 26339-26349
  • DOI: 10.1074/jbc.m115.680942
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. (2015)
  • Author(s): Takatsugu Miyazaki, Hiroyuki Yashiro, Atsushi Nishikawa and Takashi Tonozuka
  • Journal Title: Journal of Biochemistry Volume: 157 Issue: 4 Pages: 225-234
  • DOI: 10.1093/jb/mvu065
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Crystal structure and substrate-binding mode of GH63 mannosylglycerate hydrolase from Thermus thermophilus HB8. (2015)
  • Author(s): Takatsugu Miyazaki, Megumi Ichikawa, Hitoshi Iino, Atsushi Nishikawa and Takashi Tonozuka
  • Journal Title: Journal of Structural Biology Volume: 190 Issue: 1 Pages: 21-30
  • DOI: 10.1016/j.jsb.2015.02.006
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 担子菌Coprinopsis cinerea由来タンパク質CcGH131BのＸ線結晶構造解析 (2016)
  • Author(s): 奥山舜朔、林昌宏、砂川直輝、石田卓也、五十嵐圭日子、吉田誠、西河淳、殿塚隆史
  • Organizer: 日本農芸化学会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市白石区）
  • Year and Date: 2016-03-28
• [Presentation] 担子菌Coprinopsis cinerea由来機能未知タンパク質CcGH131Aの結晶構造解析 (2016)
  • Author(s): 林昌宏、奥山舜朔、田中祐太郎、田村瑞、砂川直輝、石田卓也、梅澤究、吉田誠、五十嵐圭日子、西河淳、殿塚隆史
  • Organizer: 2015年度量子ビームサイエンスフェスタ
  • Place of Presentation: つくば国際会議場（茨城県つくば市）
  • Year and Date: 2016-03-15