Study on the structure and function of glucosidases acting on glycochains

2010 Fiscal Year Final Research Report

• Project/Area Number: 20570103
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: TONOZUKA Takashi Tokyo University of Agriculture and Technology, 大学院・農学研究院, 准教授 (50285194)
• Co-Investigator(Renkei-kenkyūsha): NISHIKWA Atsushi 東京農工大学, 大学院・農学研究院, 教授 (30218127)
• Project Period (FY): 2008 – 2010
• Keywords: X線結晶解析 / 糖タンパク質糖鎖 / グルコシダーゼ

Research Abstract

The purpose of this research was to elucidate the structure-function relationship of alpha-glucosidases and related enzymes, which act on the N-linked oligosaccharides. This research also investigated the relationship between the N-linked oligosaccharides and the protein structure. The three-dimensional structure of an enzyme homologous to processing alpha-glucosidase I has been determined, and the enzyme was shown to be useful for producing oligosaccharides. The expression system of a fungal processing alpha-glucosidase I was constructed, and the enzymatic properties were determined. We also investigated some enzymatic properties of a bacterial enzyme homologous to endo-alpha-1,2-mannosidase, an enzyme potentially physiologically related to processing alpha-glucosidases. This was the first report on the enzymatic characterization of a bacterial endo-alpha-1,2-mannosidase.

Research Products

• [Journal Article] Heterologous expression, purification, and characterization of an a -mannosidase belonging to glycoside hydrolase family 99 from Shewanella amazonensis (2011)
  • Author(s): Matsuda, K., Kurakata, Y., Miyazaki, T., Matsuo, I., Ito, Y., Nishikawa, A., Tonozuka, T.
  • Journal Title: Biosci.Biotechnol.Blochem. 75 Pages: 797-799
  • Peer Reviewed
• [Journal Article] Structural similarity between a starch-hydrolyzing enzyme and an N-glycan-hydrolyzing enzyme : exohydrolases cleaving a-1, X-glucosidic linkages to produce a -glucose (2011)
  • Author(s): Tonozuka, T., Miyazaki, T., Nishikawa A.
  • Journal Title: Trends Glycosci.Glycotechnol 23 Pages: 93-102
  • Peer Reviewed
• [Journal Article] Structural insights into the substrate specificity and function of Escherichia coli K12 YgjK, a glucosidase belonging to the glycoside hydrolase family 63. (2008)
  • Author(s): Kurakata, Y., Uechi, A., Yoshida, H., Kamitori, S., Sakano, Y., Nishikawa, A., Tonozuka, T.
  • Journal Title: JMol Biol. 381 Pages: 116-128
  • Peer Reviewed
• [Presentation] Shewanella amazonenszs 由来 GH99 αMannosidase の発現系の構築と性質の解析 (2011)
  • Author(s): 松田佳奈、倉方悠馬、宮崎剛亜、松尾一郎、伊藤幸成、西河淳、殿塚隆史
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-27
• [Presentation] Molecular cloning and heterologous production of Aspergillus Niger processing a -glucosidase I (2010)
  • Author(s): 宮崎剛亜, 松本雄治, 武田陽一, 松尾一郎, 伊藤幸成, 西河淳, 殿塚隆史
  • Organizer: Plant Polysaccharide and Applied Glvcoscience Workshop2010
  • Place of Presentation: 東京(笹川記念会館)
  • Year and Date: 20100729-20100731
• [Presentation] 機能未知酵素を用いた新規な糖の創出 (2010)
  • Author(s): 殿塚隆史
  • Organizer: アグリビジネス創出フェア
  • Place of Presentation: 千葉(幕張メッセ)
  • Year and Date: 2010-11-26
• [Presentation] Structure and function of Escherichia coli YgjK, a glucosidase belonging to the glycoside hydrolase family 63, and a comparison with related enzymes (2009)
  • Author(s): 倉方悠馬, 松本雄治, 伊藤幸成, 西河淳, 殿塚隆史
  • Organizer: 8th Carbohydrate Bioengineering Meeting
  • Place of Presentation: Ischia, Naples, Italy
  • Year and Date: 20090510-20090513
• [Presentation] イソプルラナーゼの活性中心近傍に存在する残基Tyr440の役割の解明 (2009)
  • Author(s): 宮崎剛亜、矢代浩之、西河淳、殿塚隆史
  • Organizer: 日本応用糖質科学会
  • Place of Presentation: 弘前(弘前大学文京町キャンパス)
  • Year and Date: 2009-09-16
• [Presentation] Glycoside Hydrolase Family 63に属する二種類の酵素の機能解析 (2009)
  • Author(s): 倉方悠馬、松本雄治、武田陽一、松尾一郎、伊藤幸成、西河淳、殿塚隆史
  • Organizer: 日本農芸化学会
  • Place of Presentation: 福岡(マリンメッセ福岡)
  • Year and Date: 2009-03-28
• [Remarks] ホームページ等
  • URL: http://www.tuat.ac.jp/-seika/tonozuka/