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Structures and Substrate Recognitions of Animal, Botanical and Microbial α-Glucosidases and Their Molecular Evolution

Research Project

Project/Area Number 09460041
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionHokkaido University

Principal Investigator

CHIBA Seiya  Hokkaido Univ., Grad. School of Agr., Pro.90186312, 大学院・農学研究科, 教授 (30001449)

Co-Investigator(Kenkyū-buntansha) MORI Haruhide  Hokkaido Univ., Grad. School of Agr., Inst., 大学院・農学研究科, 助手 (80241363)
KIMURA Atsuo  Hokkaido Univ. Grad. School of Agr., A.Pro., 大学院・農学研究科, 助教授 (90186312)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥13,300,000 (Direct Cost: ¥13,300,000)
Fiscal Year 1999: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1998: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1997: ¥5,900,000 (Direct Cost: ¥5,900,000)
KeywordsSubstrate recognition / Carbohydrase / Catalytic site / Alpha-glucosidase / Molecular Evolution
Research Abstract

α-Glucosidase (EC 3.2.1.20) is a typical exo-type glycosidase that releases the α-glucose from non-reducing side of substrate. We are interested in the relationship between catalytic action and the structure, since the substrate specificity differs greatly with the source of enzyme. There are at least two types of α-glucosidases which show different substrate recognitions, suggesting that enzymes can be classified into two groups (family I and family II). However, the structural information is not enough to learn that structures of enzymes belonging to each group are homologous or not. In this project, we analyzed the primary structures of α-glucosidases from animal, botanical and microbial origins, and found that the enzymes of family I and II had different primary amino acid sequences.
α-Glucosidases from insect and bacteria belonged to family I, of which molecular weight was about 70 kDa. Four catalytic regions found were similar to those of α-amylase. The activity toward heteroside (sucrose or p-nitrophenyl α-glucoside) was higher than holoside (maltooligosaccharides). Members of family II were from animal, botanical and mold origins, and showed the opposite substrate specificity, high activity to holoside and low to heteroside. The molecular sizes were about 100 kDa. We analyzed the primary structures of six kinds of α-glucosidases belonging to this group. The sequences obtained were homologous each other, but no similarity was observed with family I enzymes. The findings suggest that the α-glucosidase was evolved from two different ancestral proteins.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] 木村淳夫: "Aspergillus niger結晶α-グルコシダーゼの一次構造と糖鎖"応用糖質科学. 44(2). 233-243 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 松井博和: "テンサイα-グルコシダーゼの基質特異性と全一次構造"応用糖質科学. 44(2). 245-252 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A.Kimura: "A Catalytic Amino Acid and Primary Structure of Activesite in Aspergillus niger α-Glucosidase"Biosci.Biotech.Biochem.. 61(7). 1091-1098 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S.Chiba: "Molecular Mechanism in α-Glucosidase and Glucoamylase"Biosci.Biotech.Biochem.. 61(7). 1233-1239 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N.Kitamura: "Cystein-S-conjugate β-Lyase Activity and Pyridoxal Phosphate Binding Site of Onion Alliin Lyase"Biosci.Biotech.Biochem.. 61(8). 1327-1330 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A.Kimura: "Properties and Sabstrate Specificity of α-Glucosidase from Aspergillus niger GRM3"Oyo Toshitsu Kagaku (J.Appli.Glycosci.). 44(3). 303-312 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 木村淳夫: "Aspergillus niger結晶α-グルコシダーゼの構造と機能に関する研究"応用糖質科学. 45(1). 71-79 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Nozaki,1: "Amylases and Branching Enzyme of Developing Kidney Been Seeds on Native Electrophoretic Gel"J.Appli.Glycosci.. 45(2). 117-122 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Mori: "Molecular Cloning of an α-Amylase cDNA from Germinating Cotyledons of kidney Bean (Phaseolus valgaris L.cv Toramame)"J.Appli.Glycosci.. 45(3). 261-267 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S.Igaki: "Catalytic Reaction Mechanism on α-Secondary Kinetic Isotope Effects in Hydrolytic Cleavage of α-Gukosidic Linkage by α-Glycoside"J.Appli.Glycosci.. 45(3). 269-274 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T.Saeki: "Localization of α-Glucosidase in Yeast Cells"J.Appli.Glycosci.. 45(3). 281-283 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura, T. Takayanagi, H. Mori, H. Matsui, T. Uozumi & S. Chiba: "Primary Structure and Sugar Chains of Crystalline α-Glucosidase from Asperillus niger"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 44-2. 233-243 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Matsui, I. Iwanami, H. Ito, A. Kimura, H. Mori, M. Honma & S. Chiba: "Substrate Specificity and Primary Structure of Sugar Beetα-Glucosidase"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 44-2. 245-252 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura, M. Takata, Y. Fukushi, H. Mori, H. Matsui & S. Chiba: "A Catalytic Amino Acid and Primary Structure of Active Site in Aspergillus niger α-Glucosidase"Biosci. Biotech. Biochem.. 61-7. 1091-1098 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Chiba: "Molecular Mechanism in a-Glucosidase and Glucoamylase"Biosci. Biotech. Biochem.. 61-8. 1233-1239 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kitamura, N. Shimornura, J. Iseki, M. Honma, S. Chiba, S. Tahara & J. Mizutani: "Cystein-S-conjugate β-Lyase Activity and Pyridoxal Phosphate Binding Site o Onion Alliin Lyase"Biosci. Biotech. Biochem.. 61-8. 1327-1330 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura, S. Takenaka, M. Takata, Y. Tsujisaka & S. Chib: "Properties and Substrate Specificity of α-Glucosidase from Aspergillus niger GRM3"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 44-3. 303-312 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura: "A Study on Structure and Function of Crystalline α-Glucosidase from Aspergillus niger"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 45-1. 71-79 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K. Nozaki, H. Matsui, T. Tonokawa, H. Mori, H. Ito, M. Honma & S. Chiba: "Amylases and Branching Enzyme of Developing Kidney Bean Seeds on Native Electro phoretic Gel"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 45-2. 117-122 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Mori, T. Kobayashi, T. Tonokawa, A. Tatematsu, H. Matsui, A. Kimura & S. Chiba: "Molecular Cloning of an α-Anrylase cDNA from Germinating Cotyledons of Kidney Bean (Phaseolus valgaris L. cv Toramame)"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 45-3. 261-267 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Igaki, A. Kirnura & S. Chiba: "Catalytic Reaction Mechanism on α-Secondary Kinetic Isotope Effects in Hydrolytic Cleavage of α-Glucosidic Linkage α-Glycosidases"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 45-3. 269-274 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Saeki, A. Kimura & S. Chiba: "Localization of α-Glucosidase in Yeast Cells"Oyo Toshitsu Kagaku (J. Appl. Glycosci.). 45-3. 281-283 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y. Nagasaka, N. Muraki. A. Kimura, K. Kitamoto, M. Sotou, A. Yokota & F. Tomita: "Studies on Glucomylase from Corticium rolfsii"J. Jppl. Glycosci.. 46-2. 169-178 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Mizuno, H. Mori, H. Ito, H. Matsui, A. Kimura & S. Chiba: "Molecular Cloning of Isomltotrio-dextranase Gene from Brevibacterium fuscum var. dextranlycum strain 0407 and Its Expression in Esherichia coli"Biosci. Biotech. Biochem.. 63-9. 1582-1588 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Chiba: "Studies on Molecular Mechanisms in Glycosidases"Nippon Nogeikagaku Kaishi. 73-10. 1001-1012 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura: "Immobilization of Product When Digesting Soluble Starch with Enzyme"Brain Techno News. 72-3. 27-29 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] R.S.-Jin, D. Kim, H.-J. Ryu, S. Chiba, A. Kimura & D.F. Day: "Purification and Partial Characterization of A Novel Glucanhydrolase from Lipomyces starkeyi KSM 22 and Use for Inhibition of Insoluble Glucan Formation"Biosci. Biotech. Biochem.. 64-2(in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Kimura, T. Nishio. W. Hakamada, T. Oku, G. Okada & S. Chiba: "Affinity Labeling of Glycosidase by ω-Epoxyalkyl α-glucoside"J. Jppl. Glycosci.. 47-2(in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 永坂 曜介: "Corticium rolfsiiのグルコアミラーゼに関する研究"応用糖質科学. 46(2). 169-178 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T. Mizuno: "Molecular cloning of Isomltotrio-dextranase Gene from Brevibacterium fascum var. dextranlycum strain 0407 and Its Expression in Esherichia coli"Biosci. Biotechnol. Biochem. 63(9). 1582-1588 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 千葉 誠哉: "グリコシダーゼの分子機構に関する研究"日本農芸化学会誌. 73(10). 1001-1012 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] R.S.-Jin: "Purification and partial characterization of A Novel Glucanhydrolase from Lipomyces starkey, KSM22 and Use for Inbibition of Insoluble Glucan formation"Biosci .Biotechnol. Biochem. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 木村淳夫: "ω-Epoxyalkyl α-glucosideによるグリコシダーゼの親和標識"応用糖質科学. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 木村淳夫: "澱粉粒の酵素分解に見い出された生成物の"固定化現象""BRAINテクノニュース. 72(3). 27-29 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Mori: "Molecular Cloning of an α-Amylase cDNA from Germinating Cotyledonsof Kidney Bean(Phaseolus vulgaris L.cv Toramame)" Oyo Toshitsu Kagaku(J.Appl.Glycosci.). 45・3. 261-267 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Igaki: "Catalytic Reaction Mechanism on α-Secondary Kinetic Isotope Effects in Hydrolytic Cleavage of α-Glucosidic Linkage by α-Glycosidases" Oyo Toshitsu Kagaku(J.Appl.Glycosci.). 45・3. 269-274 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Saeki: "Localization of α-Glucosidase in Yeast Cells" Oyo Toshitsu Kagaku(J.Appl.Glycosci.). 45・3. 281-283 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Nozaki: "Amylases and Branching Enzyme of Developing Kidney Bean Seeds on Native Electrophoretic Gel" Oyo Toshitsu Kagaku(J.Appl.Glycosci.). 45・2. 117-122 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Kimura: "A Study on Structure and Function of Crystalline α-Glucosidase from Aspergillus niger" Oyo Toshitsu Kagaku(J.Appl.Glycosci.). 45・1. 71-79 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Chiba: "Molecular Mechanism in α-Glucosidase and Glucoamylase" Biosci.Biotech.Biochem.61(8). 1233-1239 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Kimura: "A Catalytic Amino Acid and Primary Structure of Active Site in Aspergillus niger α-Glucosidase" Biosci.Biotech.Biochem.61(7). 1091-1098 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] R.Surarit: "Evidence for the Presence of a Single Active Site in β-D-Fucosidase/β-D-Glucosidase from Dalbergia Cochinchinensis Seeds" Biosci.Biotech.Biochem.60(8). 1265-1268 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Nakamura: "Cloning and Sequencing of an α-Glucosidase Gene from Aspergillus niger and Its Expression in A.nidulans" J.Biotechnol.53. 75-84 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Kimura: "Properties and Substrate Specificity of α-Glucosidase from Aspergillus niger GRM 3" J.Appl.Glycosci.44(3). 303-312 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Kimura: "Identification of Essential Ionizable Groups in Active Site of Aspergillus niger α-Glucosidase" Biosci.Biotech.Biocem.61(3). 475-479 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Kawagishi: "A Lectin from Mycelia of the Fungus,Ganoderma lucidum" Phytochemistry. 44(1). 7-10 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Kimura: "A Study on Structure and Function of Crystalline α-Glucosidase from Aspergillus niger" J.Appl.Glycosci.45(1),(in press). (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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