Conversion of catalytic mechanism of glycoside hydrolases

• Project/Area Number: 20780068
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: OKUYAMA Masayuki Hokkaido University, 大学院・農学研究院, 助教 (00344490)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2008: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 糖質加水分解酵素 / 触媒機構 / タンパク質工学 / 触媒メカニズム / 分子進化 / 糖転移反応

Research Abstract

I discovered an unique glycoside hydrolases family which contains inverting- and retaining-catalytic enzymes. This divergence was due to the simple difference of the position of catalytic residues. We attempted to convert the catalytic mechanism of these enzyme each other. However, the attempt was failed. These results indicated that two similar enzymes diverged from a common ancestor in early stage of evolution and each enzyme evolved in different ways.

Research Products

• [Journal Article] Catalytic reaction mechanism based on α-secondary deuterium isotope effects in hydrolysis of trehalose by European honeybee trehalase. Biosci. (2009)
  • Author(s): Mori H, Lee JH, Okuyama M, 他5名
  • Journal Title: Biotechnol. Biochem 73 Pages: 2466-2473
  • Peer Reviewed
• [Journal Article] The firstα-1, 3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31 (2009)
  • Author(s): Kang MS, Okuyama M, Mori H, Kimura A
  • Journal Title: Biochimie 91 Pages: 1434-1442
  • Peer Reviewed
• [Journal Article] Catalytic mechanism of retainingα-galactosidase belonging to glycoside hydrolase family 97. (2009)
  • Author(s): Okuyama M, Kitamura M, Hondoh H, 他5名
  • Journal Title: J. Mol. Biol. 392 Pages: 1232-1241
  • Peer Reviewed
• [Journal Article] Catalytic mechanism of retaining α-galactosidase belonging to glycoside hydrolase family 97 (2009)
  • Author(s): Okuyama M, et al
  • Journal Title: Journal of molecular biology 392 Pages: 1232-1241
  • Peer Reviewed
• [Journal Article] Catalytic reaction mechanism based on alpha-secondary deuterium isotope effects in hydrolys is of trehalose by European honeybee trehalase (2009)
  • Author(s): Mori H, et al
  • Journal Title: Biosci Biotechnol Biochem 73 Pages: 2466-2473
  • Peer Reviewed
• [Journal Article] The first α-1,3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31 (2009)
  • Author(s): Kang MS, et al
  • Journal Title: Biochimie 91 Pages: 1434-1342
  • Peer Reviewed
• [Journal Article] Structural and functional analysis of a glycoside hydrolase family 97 enzyme from Bacteroides thetaiotaomicron. (2008)
  • Author(s): Kitamura M, Okuyama M, 他7名
  • Journal Title: J. Biol. Chem. 283 Pages: 36328-36337
  • Peer Reviewed
• [Journal Article] Substrate recognition mechanism of α-1, 6-glucosidic linkage hydrolyzing enzyme, dextran glucosidase from Streptococcus mutans. (2008)
  • Author(s): Hondoh H, Saburi W, Mori H, Okuyama M, Nakada T, Matsuura Y, Kimura A
  • Journal Title: J. Mol. Biol. 378 Pages: 913-922
  • Peer Reviewed
• [Journal Article] ructural and functional analysis of a glycoside hydrolase family 97 enzyme from Bacteroides thetaiotaomicron (2008)
  • Author(s): Kitamura M et.al.
  • Journal Title: Journal of Biological Chemistry 283 Pages: 36328-36337
  • Peer Reviewed
• [Journal Article] Substrate recognition mechanism of alpha-1, 6-glucosidic linkage hydrolyzing enzy me, dextran glucosidase from Streptococcus mutans (2008)
  • Author(s): Hondoh H et.al.
  • Journal Title: Journal of Molecular Biology 378 Pages: 913-922
  • Peer Reviewed
• [Presentation] α-グリコシダーゼの機能と構造 (2010)
  • Author(s): 奥山正幸
  • Organizer: 日本農芸化学会農芸化学奨励賞受賞講演
  • Place of Presentation: 東京大学, 東京都
  • Year and Date: 2010-03-27
• [Presentation] 糖質加水分解酵素ファミリー97の配列類似性と機能類似性の関係 (2009)
  • Author(s): 奥山正幸, 他
  • Organizer: 日本農芸化学会北海道支部合同学術講演会
  • Place of Presentation: とかちプラザ, 帯広市, 北海道
  • Year and Date: 2009-11-28
• [Presentation] Bacteroides thetaiotaomicron由来SusBの触媒反応におけるCa2+の役割 (2009)
  • Author(s): 吉田拓弥, 奥山正幸, 他
  • Organizer: 応用糖質科学会21年度大会
  • Place of Presentation: 弘前大, 青森県
  • Year and Date: 2009-09-17
• [Presentation] Bacteroides thetaiotaomicron 由来SusBの触媒反応におけるCa^<2+>の役割 (2009)
  • Author(s): 吉田拓弥, 他
  • Organizer: 応用糖質科学会平成21年度大会
  • Place of Presentation: 弘前大学(青森県)
• [Presentation] Glycoside hydrolasefamily 97アノマー保持型酵素の触媒機構 (2008)
  • Author(s): 奥山正幸, 他
  • Organizer: 応用糖質科学会20年度大会
  • Place of Presentation: 琉球大, 沖縄県
  • Year and Date: 2008-09-18
• [Presentation] Functional and Structural Analysis of Glycoside Hydrolase Family 97 Enzyme from Bacteroides thetaiotaomicron (2008)
  • Author(s): Okuyama M, et, al.
  • Organizer: XXIV International Carbohydrate Symposium
  • Place of Presentation: Oslo, Norway