Establishment of novel reaction mechanism on glycosylase and its application

• Project/Area Number: 22658032
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Applied biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: KIMURA Atsuo 北海道大学, 大学院・農学研究院, 教授 (90186312)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥540,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,300,000)
• Keywords: 反応機構 / 糖質分解酵素 / 反応中間体

Research Abstract

Two reaction mechanisms have been proposed : on glycosylases, i. e. oxo-carbenium ion-mediated mechanism and covalent linkage-formation mechanism. Recently, the latter one is supported by many glycosylase-scientists. However, there is a problem in the latter mechanism : this mechanism cannot be supported by organic chemistry-related knowledge. A purpose of this project is an experimental proof of the oxo-carbenium ion-mediated mechanism. Glycosylases catalyze the useful reaction of transglycosylation. This reaction mechanism may produce the novel application approach, the possibility of which is also investigated.

Research Products

• [Journal Article] α-Glucosidase-catalyzed novel reaction on 1, 5-anhydrofructose, suggesting new metabolic pathway for production of glucose from starch (2012)
  • Author(s): Kim YM, Saburi W, Yu S, Nakai H, Maneesan J, Kang MS, Chiba S, Kim D, Okuyama M, Mori H, Kimura A
  • Journal Title: J Biol Chem Volume: (in press)
  • Peer Reviewed
• [Journal Article] Structural elucidation of dextran degradation mechanism by Streptococcus mutans dextranase belonging to glycoside hydrolase family 66 (2012)
  • Author(s): Suzuki N, Kim YM, Fujimoto Z, Momma M, Okuyama M, Mori H, Funane K, Kimura A
  • Journal Title: J Biol Chem Volume: (in press)
  • NAID: 120005183151
  • Peer Reviewed
• [Journal Article] Novel dextranase catalyzing cycloisomaltooligosaccharide-formation and its identification of catalytic amino acids and their functions using chemical rescue approach (2012)
  • Author(s): Kim YM, Kiso Y, Muraki T, Kang MS, Nakai H, Saburi W, Lang W, Kang HK, Okuyama M, Mori H, Suzuki R, Funane K, Suzuki N, Momma M, Fujimoto Z, Oguma T, Kobayashi M, Kim D, Kimura A
  • Journal Title: J Biol Chem Volume: (in press)
  • Peer Reviewed
• [Journal Article] Glycoside hydrolase family 66 enzyme homolog from Bacteroides thetaiotaomicron VPI-5482 catalyzes endo-dextranolytic and cyclization activities (2012)
  • Author(s): Kim YM, Eiji Y, Kang MS, Nakai H, Saburi W, Okuyama M, Mori H, Funane K, Kim D, Kimura A
  • Journal Title: FEBS J Volume: (in press)
  • Peer Reviewed
• [Journal Article] Biochemical characterization of a cycloisomaltooligosaccharide glucanotransferase from Paenibacillus sp. 598K (2012)
  • Author(s): Suzuki R, Terasawa K, Kimura K, Fujimoto Z, Momma M, Kobayashi M, Kimura A, Funane K
  • Journal Title: Biochim Biophys Acta Volume: (in press)
  • Peer Reviewed
• [Journal Article] Novel quantitative method for the degree of branching in dextran (2011)
  • Author(s): Kim YM, Kimura A, Kim D
  • Journal Title: Food Sci Biotechnol Volume: 20(2) Pages: 537-541
  • Peer Reviewed
• [Journal Article] Bioengineered glucansucrase of Leuconostoc mesenteroides for preferred oligosaccharide synthesis using sucrose (2011)
  • Author(s): Kang HK, Kimura A, Kim D
  • Journal Title: J Agric Food Chem Volume: (in press)
  • Peer Reviewed
• [Journal Article] Truncation of N-and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity (2011)
  • Author(s): Kim YM, Shimizu R, Nakai H, Mori H, Okuyama M, Kang MS, Fujimoto Z, Funane K, Kim D, Kimura A
  • Journal Title: Appl Microbiol Biotechnol Volume: (in press)
  • NAID: 120003811531
  • Peer Reviewed
• [Journal Article] Calcium ion-dependent increase in thermostability of dextran glucosidase from Streptococcus mutans (2011)
  • Author(s): Kobayashi M, Hondoh H, Mori H, Saburi W, Okuyama M, Kimura A
  • Journal Title: Biosci Biotechnol Biochem Volume: 75(8) Pages: 1557-1563
  • NAID: 10029591699
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary crystallographic analysis of dextranase from Streptococcus mutans (2011)
  • Author(s): Suzuki N, Kim YM, Fujimoto Z, Momma M, Kang HK, Funane K, Okuyama M, Mori H, Kimura A
  • Journal Title: Acta Crystallogr F Volume: F67 Pages: 1542-1544
  • NAID: 120003660602
  • Peer Reviewed
• [Journal Article] Function of C-terminal region of cycloisomaltooligosaccharide glucanotransferase from Bacillus circulams T-3040 (2011)
  • Author(s): Funane K, Kawabata Y, Kim YM, Kang HK, Fujimoto Z, Kimura A, Kobayashi M
  • Journal Title: Biochim Biophys Acta Volume: 1814 Pages: 428-434
  • Peer Reviewed
• [Journal Article] 環状イソマルトオリゴ糖グルカノトランスフェラーゼの分子解析とBacillus circulans T-3040株における環状イソマルトオリゴ糖の新規な生合成経路の発見 (2011)
  • Author(s): 舟根和美, 川端康之, 鈴木龍一郎, 藤本瑞, 北岡本光, 木村淳夫, 小林幹彦
  • Journal Title: 応用糖質科学 Volume: 1 Pages: 179-185
  • Peer Reviewed
• [Journal Article] Novel quantitative method for the degree of branching in dextran (2011)
  • Author(s): Kim YM
  • Journal Title: Food Sci Biotechnol Volume: 20 Pages: 537-541
  • Peer Reviewed
• [Journal Article] Bioengineering of Leuconostoc mesenteroides glucansucrases that gives selected bond formation for glucan synthesis and/or acceptor-product synthesis (2011)
  • Author(s): Kang HK, Kimura A, Kim D
  • Journal Title: J Agric Food Chem Volume: 59(8) Issue: 8 Pages: 4148-4155
  • DOI: 10.1021/jf104629g
  • Peer Reviewed
• [Journal Article] Truncation of N- and C-terminal regions of Streptococcus mutans dextranase enhances catalytic activity (2011)
  • Author(s): Kim YM
  • Journal Title: Appl Microbiol Biotechnol Volume: 91 Issue: 2 Pages: 329-339
  • DOI: 10.1007/s00253-011-3201-y
  • Peer Reviewed
• [Journal Article] Molecular analysis of cycloisomaltooligosaccharide glucanotransferase and discovery of novel biosynthetic pathway of cycloisomaltooligosaccharide in Bacillus circulams T-3040 (2011)
  • Author(s): Funane K
  • Journal Title: Bull Appl Glycosci Volume: 1 Pages: 179-185
  • Peer Reviewed
• [Journal Article] Calcium Ion-Dependent Increase in Thermostability of Dextran Glucosidase from Streptococcus mutans (2011)
  • Author(s): Kobayashi M
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 75 Issue: 8 Pages: 1557-1563
  • DOI: 10.1271/bbb.110256
  • NAID: 10029591699
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Suicide substrate-based inactivation of endodextranase byω-epoxyalkylα-D-glucopyranosides (2010)
  • Author(s): Kang HK, Kim YM, Nakai H, Kang MS, Hakamada W, Okuyama M, Mori H, Nishio T, Kimura A
  • Journal Title: J Appl Glycosci Volume: 57 Pages: 111-117
  • Peer Reviewed
• [Journal Article] Optimal expression and characterization of a fusion enzyme having dextransucrase and dextranase activities (2010)
  • Author(s): Ryu HJ, Jin X, Lee JH, Woo HJ, Kim YM, Kim GJ, Seo ES, Kang HK, Kim J, Cho DL, Kimura A, Kim D
  • Journal Title: Enzyme Microb Technol Volume: 47 Pages: 212-215
  • Peer Reviewed
• [Journal Article] Rice Os4BGlu12 is a wound-inducedβ-glucosidase that hydrolyzes cell-wall-β-glucan-derived oligosaccharides and glycosides (2010)
  • Author(s): Opassiri R, Maneesan J, Akiyama T, Pomthong B, Jin S, Kimura A, Ketudat-Cairns JR
  • Journal Title: Plant Sci Volume: 179 Pages: 273-280
  • Peer Reviewed
• [Presentation] 糖質酵素の分子機構に関する研究 (2012)
  • Author(s): 木村淳夫
  • Organizer: 日本応用糖質科学会平成24年度大会(第61回)
  • Place of Presentation: 東京農工大学(東京都)
  • Year and Date: 2012-09-20
• [Presentation] 摂取するイソマルト糖の重合度はデキストラン硫酸ナトリウムによる大腸炎症状に影響する (2012)
  • Author(s): 岩谷一史
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-24
• [Presentation] 直鎖イソマルトメガロ糖はラット小腸におけるケルセチン配糖体吸収を促進する (2012)
  • Author(s): 篠木亜季
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-24
• [Presentation] Streptococcus mutans由来デキストラナーゼのX線結晶構造解析 (2012)
  • Author(s): 鈴木喜大
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-23
• [Presentation] 環状イソマルトオリゴ糖グルカノトランスフェラーゼの活性向上と生成物サイズ特異性の制御 (2012)
  • Author(s): 鈴木龍一郎
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-23
• [Presentation] デキストラナーゼに見出された『プロ酵素を介する活性化現象』の分子機構 (2012)
  • Author(s): キムヨンミン
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-23
• [Presentation] Functional implication of amino acid residues in conserved region II of European honeybee α-glucosidase on substrate specificity, transglucosylation and substrate inhibition (2012)
  • Author(s): Lukana Ngiwsara
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-23
• [Presentation] Bacteroides thetaiotaomicron由来α-galactosidaseの求核残基変異酵素が触媒する糖転移反応の解析 (2012)
  • Author(s): 渡辺健一
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大(京都市)
  • Year and Date: 2012-03-23
• [Presentation] Streptococcus mutans由来デキストラナーゼの結晶構造に基づく触媒機構の解明 (2012)
  • Author(s): 鈴木喜大
  • Organizer: 第29回PFシンポジウム
  • Place of Presentation: つくば国際会議場(つくば市)
• [Presentation] 環状イソマルトオリゴ糖グルカノトランスフェラーゼの合目的変異導入による生成物特異性の制御 (2011)
  • Author(s): 鈴木龍一郎,鈴木喜大,藤本瑞,門間充,木村啓太郎,木村淳夫,舟根和美
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)ポスター賞受賞
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] 環状イソマルトオリゴ糖グルカノトランスフェラーゼの合目的変異導入による生成物特異性の制御 (2011)
  • Author(s): 鈴木龍一郎
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] 環状イソマルトオリゴ糖グルカノトランスフェラーゼのβ→αループ2はCI-8の優先的生成に寄与する (2011)
  • Author(s): 齊藤みどり
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] Isomaltooligosaccharide 6-α-glucosyltransferaseサブサイト+1変異酵素の機能解析 (2011)
  • Author(s): 青山泰
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] アノマー反転型トレハラーゼ変異体により合成された新規非還元二糖の構造 (2011)
  • Author(s): 平内亨
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] Production and investigation of recombinant α-glucosidase from Podospora anserina (2011)
  • Author(s): Kyung-Mo Song
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] GH family 31に属するAspergillus niger由来のα-glucosidaseの鎖長特異性の改変 (2011)
  • Author(s): 田上貴祥
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] Dextran glucosidase (DexB)ウォータパス改変体の構造と機能 (2011)
  • Author(s): 小林桃子
  • Organizer: 日本応用糖質科学会平成23年度大会(第60回)
  • Place of Presentation: 北大(札幌市)
  • Year and Date: 2011-09-28
• [Presentation] イソマルトメガロ糖とその生産酵素 (2011)
  • Author(s): 木村淳夫
  • Organizer: メガロ糖・第2回シンポジウム「メガロ糖の製造と機能性ナノ材料:イソマルト型の新展開」
  • Place of Presentation: 帯畜大(帯広市)
  • Year and Date: 2011-09-16
• [Presentation] イソマルトメガロ糖とその生産酵素 (2011)
  • Author(s): 木村淳夫
  • Organizer: メガロ糖・第2回シンポジウム「メガロ糖の製造と機能性ナノ材料:イソマルト型の新展開」
  • Place of Presentation: 帯畜大(帯広市)(招待講演)
  • Year and Date: 2011-09-16
• [Presentation] Molecular anatomy of α-1,6-glucoside(s)-forming and -hydrolyzing enzymes, comprising megalosaccharide-producing enzymes (2011)
  • Author(s): Young-Min Kim
  • Organizer: 2011 CAB Agricultural Biotechnology Symposium on "Agricultural Biomaterials for Next Generation"
  • Place of Presentation: ソウル大学(ソウル市、韓国)(招待講演)
  • Year and Date: 2011-08-25
• [Presentation] Bacillus circulans T-3040由来サイクロデキストラン合成酵素と環状イソマルトオリゴ糖との複合体のX線結晶構造解析 (2011)
  • Author(s): 鈴木喜大
  • Organizer: 第28回PFシンポジウム
  • Place of Presentation: つくば国際会議場(つくば市)
  • Year and Date: 2011-07-12
• [Presentation] Molecular mechanism of novel enzymes degrading and formingα-1, 6-glucosidic linkages, including megalosaccharide production (2011)
  • Author(s): Kim YM, Kanegae M, Hondoh H, Nishimura T, Saburi W, Sadahiro J, Lang W, Shimizu R, Nakai H, Okuyama M, Mori H, Kang MS, Fujimoto Z, Funane K, Kim D, A Kimura A
  • Organizer: The 78th Annual Meeting and International Symposium in 2011 Annual Meeting of Korean Society of Food Science and Technology(KoSFoST)
  • Place of Presentation: Daegu Exhibition Convention Center(Daegu Metropolitan City, Kora)
  • Year and Date: 2011-06-09
• [Presentation] Molecular mechanism of novel enzymes degrading and forming α-1,6-glucosidic linkages, including megalosaccharide production (2011)
  • Author(s): Kim YM
  • Organizer: The 78th Annual Meeting and International Symposium in 2011 Annual Meeting of Korean Society of Food Science and Technology (KoSFoST)
  • Place of Presentation: Daegu Exhibition Convention Center(大邱広域市、韓国)(招待講演)
  • Year and Date: 2011-06-09
• [Presentation] Dietary influence of linearα-1, 6-polysaccharides on several parameters under experimental colitis depends on degree of the polymerization (2011)
  • Author(s): Hitoshi Iwaya, Jae-Sung Lee, Shinya Yamagishi, Aki Shinoki, Weeranuch Lang, Hee-Kwon Kang, Hiroshi Hara, Atsuo Kimura, Satoshi Ishizuka
  • Place of Presentation: Royton Sapporo Hotel(Sapporo, Japan)
• [Presentation] Structural element determining the diverse specificities for chain-length of substrate in glycoside hydrolase family 31α-glucosidases (2011)
  • Author(s): Tagami T, Okuyama M, Mori H, Kimura A
  • Organizer: 9th Carbohydrate Bioengineering Meeting
  • Place of Presentation: Calouste Gulbenkian Foundation(Lisbon, Portugal)
• [Presentation] Enhancement of GH66 dextranase activity by truncation of N- and C-terminal variable regions, suggesting proenzyme-like activation (2011)
  • Author(s): Mori H
  • Organizer: 9th Carbohydrate Bioengineering Meeting
  • Place of Presentation: Calouste Gulbenkian Foundation (Lisbon, Portugal)
• [Presentation] Structural element determining the diverse specificities for chain-length of substrate in glycoside hydrolase family 31 α-glucosidases (2011)
  • Author(s): Tagami T
  • Organizer: 9th Carbohydrate Bioengineering Meeting
  • Place of Presentation: Calouste Gulbenkian Foundation (Lisbon, Portugal)
• [Presentation] Alteration of product specificity of cycloisomaltooligosaccharide glucanotransferase from Bacillus circulans T-3040 (2011)
  • Author(s): Suzuki R
  • Organizer: 16th European Carbohydrate Symposium
  • Place of Presentation: Hilton Sorrento Palace Congress Centre (Naples, Italy)
• [Presentation] Dietary influence of linear α-1,6-polysaccharides on several parameters under experimental colitis depends on degree of the polymerization (2011)
  • Author(s): Hitoshi Iwaya
  • Organizer: International Society for Nutraceuticals and Functional Foods
  • Place of Presentation: ロイトン札幌(札幌市)
• [Presentation] 糖質酵素の基礎研究とその展開 (2010)
  • Author(s): 木村淳夫
  • Organizer: 日本応用糖質科学会平成22年度大会
  • Place of Presentation: グランシップ(静岡県)
  • Year and Date: 2010-09-17
• [Presentation] Molecular anatomy ofα-1, 6-glucoside(s)-forming and-hydrolyzing enzymes, comprising megalosaccharide-producing enzymes (2010)
  • Author(s): Young-Min Kim, Michiyo Kanegae, Hironori Hondoh, Takashi Nishimura, Wataru Saburi, Juri Sadahiro, Weeranuch Lang, Ryoko Shimizu, Hiroyuki Nakai, Hee-Kwon Kang, Masayuki Okuyama, Haruhide Mori, Min-Sun Kang, Zui Fujimoto, Kazumi Funane, Doman Kim, Kimura A
  • Organizer: 2011 CAB Agricultural Biotechnology Symposium on "Agricultural Biomaterials for Next Generation"
  • Place of Presentation: Seoul Natinal University(Seoul, Korea)
  • Year and Date: 2010-08-25
• [Presentation] GH family 31における基質の鎖長認識機構の解明 (2010)
  • Author(s): 田上貴祥
  • Organizer: 日本農芸化学会北海道支部夏季シンポジウム
  • Place of Presentation: とかちプラザ(帯広市)
  • Year and Date: 2010-08-06
• [Presentation] Dextran glucosidase and isomaltooligosaccharide 6-α-glucosyl transferase : mechanism of substrate recognition and transglucosylation (2010)
  • Author(s): Mori H, Hondoh H, Saburi W, Otsuka-Rachi H, Nishimura T, Kanegae M, YM Kim, Okuyama M, Kimura A
  • Organizer: 4th Symposium of theα-Amylase Family
  • Place of Presentation: Smolenice Castle, Slovakia
• [Presentation] Putative role of Tyr227, Gln349 and Leu350 near catalytic amino acids of honeybeeα-glucosidase III (2010)
  • Author(s): Ngiwsara L, Iwai G, Mori H, Okuyama M, Kimura A
  • Organizer: 4th Symposium on the Alpha-Amylase Family
  • Place of Presentation: Smolenice Cattle(Smolenice, Slovakia)
• [Remarks] アウトリーチ活動情報奥山正幸、木村淳夫:独立行政法人科学技術振興機構・平成23年度サイエンス・パートナーシップ・プロジェクト(SPP)事業(北海道札幌藻岩高校)アウトリーチ活動情報
• [Remarks] 奥山正幸、木村淳夫:独立行政法人科学技術振興機構・平成22年度サイエンス・パートナーシップ・プロジェクト(SPP)事業(北海道札幌藻岩高校)アウトリーチ活動情報
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• [Remarks]
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