Elucidation of catalytic mechanisms of thermostable glycoside-hydrolases by structural analyses

• Project/Area Number: 25450143
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: MIne Shouhei 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 主任研究員 (70415751)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: グルコサミン / 好熱菌 / 酵素 / バイオマス / キチン / 耐熱性 / X線結晶構造解析 / 古細菌

Outline of Final Research Achievements

Chitin is a polysaccharide that comprises a major constituent of fungal cell walls, the exoskeletons of insects, and the shells of crustaceans. N-acetylglucosamine (GlcNAc) and Glucosamine (GlcN), which is derived from the hydrolysis of chitin or chitosan,have a variety of biological functions and have therefore been used as a food additive and in medicines. Using the chitin catabolic pathway of hyperthemophilie,we have focused two thermostable enzymes, deacetylase (Dac) (EC 3.5.1.-) and β-N-acetylglucosaminidases (NagA) (EC 3.2.1.52). To understand the catalysis and adaptation to extreme high temperature of these enzymes, we have determined the structures of Dac and NagA.

Research Products

• [Journal Article] A tilt-pair based method for assigning the projection directions of randomly oriented single-particle molecules (2015)
  • Author(s): Yutaka Ueno, Shouhei Mine and Kazunori Kawasaki
  • Journal Title: Microscopy Volume: 64 Issue: 2 Pages: 129-141
  • DOI: 10.1093/jmicro/dfv002
  • Peer Reviewed
• [Journal Article] Expression from engineered Escherichia coli chromosome and crystallographic study of archaeal N,N'-diacetylchitobiose deacetylase (2014)
  • Author(s): Shouhei Mine, Mayumi Niiyama, Wakana Hashimoto, Takahisa Ikegami, Daisuke Koma, Takashi Ohmoto, Yohta Fukuda, Tsuyoshi Inoue, Yoshito Abe, Tadashi Ueda, Junji Morita, Koichi Uegaki and Tsutomu Nakamura
  • Journal Title: FEBS JOURNAL Volume: 281 Issue: 11 Pages: 2584-2596
  • DOI: 10.1111/febs.12805
  • Peer Reviewed
• [Journal Article] The structure of hyperthermophilic β-N-acetylglucosaminidase reveals a novel dimer architecture associated with the active site. (2014)
  • Author(s): Shouhei Mine, Yuji Kado, Masahiro Watanabe, Yohta Fukuda, Yoshito Abe, Tadashi Ueda, Yutaka Kawarabayasi, Tsuyoshi Inoue and Kazuhiko Ishikawa
  • Journal Title: FEBS JOURNAL Volume: 281 Issue: 22 Pages: 5092-5103
  • DOI: 10.1111/febs.13049
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Solution structure of the chitin-binding domain 1 (ChBD1) of a hyperthermophilic chitinase from Pyrococcus furiosus (2014)
  • Author(s): Shouhei Mine, Tsutomu Nakamura, Takaaki Sato, Takahisa Ikegami, and Koichi Uegaki
  • Journal Title: The Journal of Biochemistry Volume: 155 Pages: 115-122
  • NAID: 40019985734
  • Peer Reviewed