Conversion of hydrolases into transglycosidases

• Project/Area Number: 22580099
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: MORI Haruhide 北海道大学, 大学院・農学研究院, 准教授 (80241363)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 酵素化学 / 糖転移 / 酵素反応機構 / オリゴ糖 / イソマルトオリゴ糖

Research Abstract

A variety of oligosaccharides can be produced by carbohydrate hydrolases. In this work, amino acid residues responsible for high transglycosylation activities were identified through the comparison of transglycosidases with hydrolases. It was clarified that one transglycosidase had a catalytic domain similar to anomer-inverting hydrolases, which possess no transglycosidase activity.

Research Products

• [Journal Article] Enzymatic Synthesis of Acarviosyl-maltooligosaccharides Using Disproportionating Enzyme 1 (2013)
  • Author(s): Tagami T, Tanaka Y, Mori H, Okuyama M, and Kimura A
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 77 Issue: 2 Pages: 312-319
  • DOI: 10.1271/bbb.120732
  • NAID: 10031164734
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Amino Acids in Conserved Region II Are Crucial to Substrate Specificity, Reaction Velocity, and Regioselectivity in the Transglucosylation of Honeybee GH-13 α-Glucosidases (2012)
  • Author(s): Tagami T, Tanaka Y, Okuyama M, Mori H, Kimura A
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 76 Issue: 10 Pages: 1967-1974
  • DOI: 10.1271/bbb.120473
  • NAID: 130004054024
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [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: 287(27) Issue: 27 Pages: 22441-22444
  • DOI: 10.1074/jbc.c112.360909
  • 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
  • Journal Title: J Biol Chem Volume: 287 Issue: 24 Pages: 19927-19935
  • DOI: 10.1074/jbc.m111.339036
  • 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: 287(24) Issue: 24 Pages: 19916-19926
  • DOI: 10.1074/jbc.m112.342444
  • NAID: 120005183151
  • 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 Issue: 12 Pages: 1542-1544
  • DOI: 10.1107/s1744309111038425
  • NAID: 120003660602
  • Peer Reviewed
• [Journal Article] Transglycosylation by barley α -amylase 1. (2011)
  • Author(s): Motyana JA, Fazekas E, Mori H, Svensson B, Bagossi P, Kandra L, and Gyemant G.
  • Journal Title: J. Mol. Catalysis B, Enzymatic Volume: 72 Issue: 3-4 Pages: 229-237
  • DOI: 10.1016/j.molcatb.2011.06.010
  • 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, and Kimura A.
  • Journal Title: Biosci. Biotechnol. Biochem Volume: 75(8) Pages: 1557-1563
  • NAID: 10029591699
  • 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] 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] 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: (印刷中)
  • Peer Reviewed
• [Journal Article] Suicide Substrate-based Inactivation of Endodextranase by .OMEGA.-Epoxyalkyl .ALPHA.-D-Glucopyranosides (2010)
  • Author(s): Kang HK, Kim YM, Nakai H, Kang MS, Hakamada W, Okuyama M, Mori H, Nishio T, and Kimura A.
  • Journal Title: Journal of Applied Glycoscience Volume: 57 Issue: 4 Pages: 269-272
  • DOI: 10.5458/jag.57.269
  • NAID: 10026967780
  • ISSN: 1344-7882, 1880-7291
  • Peer Reviewed
• [Journal Article] Suicide Substrate-based Inactivation of Endodextranase by ω-Epoxyalkyl α-D-Glucopyranosides. (2010)
  • Author(s): Kang, H-K
  • Journal Title: J.Appl.Glycosci. Volume: 57 Pages: 269-272
  • Peer Reviewed
• [Presentation] Conversion of Trehalose Hydrolase into Oligosaccharide Synthase by Mutation around theCatalytic Amino Acid". (2012)
  • Author(s): Hirauchi, T., Mori, H.
  • Organizer: Conference on Agricultural Biodiversity and Sustainability 2012 (CABS2012)
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2012-08-29
• [Presentation] Identification of amino acids responsible for the substrate specificity in a novelα-glucosyltransferase from Bacillus species". (2012)
  • Author(s): Aoyama, Y., Mori, H.
  • Organizer: Conference on Agricultural Biodiversity and Sustainability 2012 (CABS2012)
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2012-08-29
• [Presentation] Enzymatic synthesis of acarviosyl-maltooligosaccha rides and their inhibitory effects on α-glucosidase (2012)
  • Author(s): Tagami, T., Mori, H.
  • Organizer: 第26回 International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla(スペイン)
  • Year and Date: 2012-07-24
• [Presentation] Amino acid residues in conserved region II involved in specific function of European honeybee(Apis mellifera) α-glucosidase isozymes". (2012)
  • Author(s): Ngiwsara, L., Mori, H.
  • Organizer: 第26回 International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla(スペイン)
• [Presentation] Domain organization and catalytic amino acids of dextran dextrinase: structural similarity to inverting glucodextanase (or glucoamylase) in catalytic domain". (2012)
  • Author(s): Sadahiro, J., Mori, H.
  • Organizer: 第26回 International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla(スペイン)
• [Presentation] Identification of a key residue for regioselectivity of α-glucosidase from Schwanniomycesoccidentalis". (2012)
  • Author(s): Song, K.M., Mori, H.
  • Organizer: 第26回 InternationalCarbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla(スペイン)
• [Presentation] Production and characterization of recombinantα-glucosidase from Podospora anserina which reveals high regioselectivity forα-1,3-glucosidic linkage". (2012)
  • Author(s): Song, K.M., Mori, H.
  • Organizer: Plant andSeaweed Polysaccharides Symposium
  • Place of Presentation: La Cite, Nantes events center (フランス)
• [Presentation] Production and characterization of recombinant α-glucosidase from Podospora anserina which reveals high regioselectivity for α-1,3-glucosidic linkage (2012)
  • Author(s): Kyung-Mo Song
  • Organizer: Plant and Seaweed Polysaccharides Symposium
  • Place of Presentation: La Cite, Nantes events center (フランス)
• [Presentation] Identification of a key residue for regioselectivity of α-glucosidase from Schwanniomyces occidentalis (2012)
  • Author(s): Kyung-Mo Song
  • Organizer: 第26回International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla（スペイン）
• [Presentation] Domain organization and catalytic amino acids of dextran dextrinase: structural similarity to inverting glucodextanase (or glucoamylase) in catalytic domain (2012)
  • Author(s): Sadahiro J
  • Organizer: 第26回International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla（スペイン）
• [Presentation] Amino acid residues in conserved region II involved in specific function of European honeybee (Apis mellifera) α-glucosidase isozymes (2012)
  • Author(s): Ngiwsara L
  • Organizer: 第26回International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla（スペイン）
• [Presentation] 加水分解酵素を用いたオリゴ糖合成 (2011)
  • Author(s): 森春英
  • Organizer: 第1回新潟大学国際糖質科学シンポジウム「糖質科学の最前線
  • Place of Presentation: 新潟大学(新潟)
  • Year and Date: 2011-10-03
• [Presentation] 加水分解酵素を用いたオリゴ糖合成 (2011)
  • Author(s): 森春英
  • Organizer: 第1回新潟大学国際糖質科学シンポジウム「糖質科学の最前線」
  • Place of Presentation: 新潟大学(新潟)(招待講演)
  • Year and Date: 2011-10-03
• [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,森春英
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] Isomaltooligosaccharide 6-α-glucosyl transferaseサブサイト+1変異酵素の機能解析 (2011)
  • Author(s): 青山泰,森春英
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] Dextran glucosidase (DexB)ウォーターパス改変体の機能と構造 (2011)
  • Author(s): 小林桃子,森春英
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] Functional implication of amino acid residues in conserved region II of European honeybee α-glucosidase on substrate specificity, transglucosylation and substrate inhibition. (2011)
  • Author(s): Lukana, N., Mori, H.
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • 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H23大会
  • 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β→αループ2はCI-8の優先的生成に 寄与する (2011)
  • Author(s): 藤みどり
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] Functional implication of amino acid residues in conserved region II of European honeybee α-glucosidase on substrate specificity, transglucosylation and substrate inhibition (2011)
  • Author(s): Lukana Ngiwsara
  • Organizer: 日本応用糖質科学会H23大会
  • Place of Presentation: 北海道大学(札幌)
  • Year and Date: 2011-09-28
• [Presentation] Dextran glucosidase (DexB)のウォーターパス改変酵素の機能解析 (2011)
  • Author(s): 小林桃子
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [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が示すCI-8の優先的生成に寄与する構造因子 (2011)
  • Author(s): 齊藤みどり
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [Presentation] ホタテガイ(Pastinopecton yessoensis)中腸腺由来α-glucosidase-Cl^-による反応速度の上昇と基質特異性の変化- (2011)
  • Author(s): 桝田安志
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [Presentation] アノマー反転型トレハラーゼ変異体によるオリゴ糖合成 (2011)
  • Author(s): 平内亨
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [Presentation] Ruminococcus albus NE1のマンナン代謝の解析 (2011)
  • Author(s): 中島碧
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [Presentation] Ruminococcus albus NE1株由来cellobiose phosphorylaseの一次構造の解析と大腸菌による組換え酵素の諸性質 (2011)
  • Author(s): 阿部正太郎
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都(京都女子大学)
  • Year and Date: 2011-03-26
• [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: 第9回Carbohydrate Bioengineering Meeting
  • Place of Presentation: the CalousteGulbenkian Foundation(ポルトガル)
• [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: 第9回Carbohydrate Bioengineering Meeting
  • Place of Presentation: リスボン(ポルトガル)
• [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: 第9回Carbohydrate Bioengineering Meeting
  • Place of Presentation: リスボン(ポルトガル)
• [Presentation] Dextran glucosidase and isomaltooligosaccharide 6-α-glucosyl transferase : mechanism of substrate recognition and transglucosylation. (2010)
  • Author(s): Mori, H.
  • Organizer: 4th symposium on alpha-amylase family.
  • Place of Presentation: Smolenice Castle(スロバキア) 招待講演
  • Year and Date: 2010-09-28
• [Presentation] 加水分解反応の抑制により縮合反応の生成物を蓄積させるグルコアミラーゼ変異体 (2010)
  • Author(s): 砂守このみ,森春英
  • Organizer: 日本農芸化学会東北・北海道支部合同講演会
  • Place of Presentation: 東北大農学部(仙台)
  • Year and Date: 2010-09-28
• [Presentation] Dextran glucosidase and isomaltooligosaccharide 6-α-glucosyltransferase : mechanism of substrate recognition and transglucosylation. (2010)
  • Author(s): Mori, H.
  • Organizer: 4th symposium on alpha-amylase family
  • Place of Presentation: Smolenice Castle(スロバキア) 招待講演
  • Year and Date: 2010-09-28
• [Presentation] Putative role of Tyr227, Gln349 and Leu350 near catalytic amino acids of honeybee α-glucosidase III. (2010)
  • Author(s): Ngiwsara, L.
  • Organizer: 4th symposium on alpha-amylase family
  • Place of Presentation: Smolenice Castle(スロバキア)
  • Year and Date: 2010-09-28
• [Presentation] 加水分解反応の抑制により縮合反応の生成物を蓄積させるグルコアミラーゼ変異体 (2010)
  • Author(s): 砂守このみ
  • Organizer: 日本農芸化学会東北・北海道支部合同講演会
  • Place of Presentation: 仙台(東北大農学部)
  • Year and Date: 2010-09-28
• [Presentation] Oligo-1, 6-glucosidaseがCl-依存性を示すために必要なアミノ酸残基の決定 (2010)
  • Author(s): 寺田智明
  • Organizer: 日本農芸化学会東北・北海道支部合同講演会
  • Place of Presentation: 仙台(東北大農学部)
  • Year and Date: 2010-09-28
• [Presentation] isomaltooligo saccharide 6-α-glucosyl transferase のサブサイト+1変異酵素の機能解析 (2010)
  • Author(s): 西村崇志,森春英
  • Organizer: 日本応用糖質科学会
  • Place of Presentation: グランシップ(静岡)
  • Year and Date: 2010-09-15
• [Presentation] isomaltooligosaccharide 6-α-glucosyltransferaseのサブサイト+1変異酵素の機能解析 (2010)
  • Author(s): 西村崇志
  • Organizer: 日本応用糖質科学会
  • Place of Presentation: 静岡市グランシップ
  • Year and Date: 2010-09-15
• [Presentation] Aspergillus niger α-glucosidaseのサブサイト+1の改変 (2010)
  • Author(s): 田上貴祥
  • Organizer: 日本応用糖質科学会
  • Place of Presentation: 静岡市グランシップ
  • Year and Date: 2010-09-15
• [Presentation] Glycosynthase Reaction Catalyzed by Trehalase Mutant Enzymes (2010)
  • Author(s): Mori, H.
  • Organizer: The 25^<th> International Carbohydrate Symposium
  • Place of Presentation: 千葉市幕張メッセ
  • Year and Date: 2010-08-05
• [Presentation] Effects on reactivity of honeybee α-glucosidase III mutants by site-direct mutagenesis of residues adjacent to highly conserved region II and IV. (2010)
  • Author(s): Ngiwsara, L., Mori, H.
  • Organizer: The 25th International Carbohydrate Symposium
  • Place of Presentation: 幕張メッセ(千葉)
  • Year and Date: 2010-08-02
• [Presentation] Glycosynthase Reaction Catalyzed by Trehalase Mutant Enzymes (2010)
  • Author(s): Mori, H.
  • Organizer: The 25th International Carbohydrate Symposium.
  • Place of Presentation: 幕張メッセ(千葉)
  • Year and Date: 2010-08-02
• [Presentation] Effects on reactivity of honeybee α-glucosidase III mutants by site-direct mutagenesis of residues adjacent to highly conserved region II and IV (2010)
  • Author(s): Ngiwsara, L.
  • Organizer: The 25^<th> International Carbohydrate Symposium
  • Place of Presentation: 千葉市幕張メッセ
  • Year and Date: 2010-08-02
• [Presentation] Effects of mutation of His-541 and Glu-542 on catalytic activities of cycloisomaltooligosaccharide glucanotransferase from Bacillus circulans T-3040 (2010)
  • Author(s): Kang, HK.
  • Organizer: The 25^<th> International Carbohydrate Symposium
  • Place of Presentation: 千葉市幕張メッセ
  • Year and Date: 2010-08-02
• [Presentation] Shifting specificity of Aspergillus niger α-glucosidase from short-chain substrates to long-chain substrates (2010)
  • Author(s): Tagami, T.
  • Organizer: Plant Polysaccharide Workshop and Applied Glycoscience
  • Place of Presentation: 東京笹川ホール
  • Year and Date: 2010-07-31
• [Presentation] デキストラン合成酵素dextran dextrinaseの触媒残基およびその機能の決定 (2010)
  • Author(s): 貞廣樹里,森春英
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学(仙台)
• [Presentation] Structural insights into specificity for long-chain substrates of sugar beet α-glucosidase.
  • Author(s): Takayoshi Tagami
  • Organizer: Plant and Seaweed Polysaccharides Symposium
  • Place of Presentation: La Cite, Nantes events center (フランス)
• [Presentation] Enzymatic synthesis of acarviosyl-maltooligosaccharides and their inhibitory effects on α-glucosidase
  • Author(s): Tagami, T
  • Organizer: 第26回International Carbohydrate Symposium
  • Place of Presentation: Hotel Melia Castilla（スペイン）
• [Presentation] Identification of amino acids responsible for the substrate specificity in a novel α-glucosyltransferase from Bacillus species.
  • Author(s): Aoyama Y
  • Organizer: Conference on Agricultural Biodiversity and Sustainability 2012 (CABS2012)
  • Place of Presentation: 北海道大学（札幌）
• [Presentation] Conversion of Trehalose Hydrolase into Oligosaccharide Synthase by Mutation around the Catalytic Amino Acid
  • Author(s): Hirauchi, T
  • Organizer: Conference on Agricultural Biodiversity and Sustainability 2012 (CABS2012)
  • Place of Presentation: 北海道大学（札幌）
• [Presentation] Acarviosyl-maltooligosaccharideの酵素合成とテンサイα-glucosidaseに対する基質アナログとしての利用
  • Author(s): 田上 貴祥
  • Organizer: 日本応用糖質科学会H24大会
  • Place of Presentation: 東京農工大(東京・府中）
• [Presentation] デキストラン合成酵素dextran dextrinaseの触媒残基およびその機能の決定
  • Author(s): 貞廣 樹里
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学（仙台）
• [Presentation] Purification and characterization of nigerose-producing enzyme from Podospora anserina
  • Author(s): Kyung-Mo SONG
  • Organizer: 日本農芸化学会2013年度大会
  • Place of Presentation: 東北大学（仙台）