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2000 Fiscal Year Final Research Report Summary

Molecular and cellular studies on biofunctional proteins from microorganisms.

Research Project

Project/Area Number 10306007
Research Category

Grant-in-Aid for Scientific Research (A).

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

Principal Investigator

KUMAGAI Hidehiko  Kyoto University, Graduate School of Biostudies, Professor, 生命科学研究科, 教授 (70027192)

Co-Investigator(Kenkyū-buntansha) TAMAKI Hisanori  Kyoto University, Graduate School of Biostudies, Assistant, 生命科学研究科, 助手 (20212045)
SUZUKI Hideyuki  Kyoto University, Graduate School of Biostudies, Associate Professor, 生命科学研究科, 助教授 (10202136)
YAMAMOTO Kenji  Kyoto University, Graduate School of Biostudies, Professor, 生命科学研究科, 教授 (70109049)
Project Period (FY) 1998 – 2000
Keywordsglycopeptide / glycosidase / transglycosidation / glutathione / aminopeptidase / γ-glutamyltranspeptidase / cAMP / pseudohyphal growth
Research Abstract

1. We analyzed the function of specific glycosidases produced by microorganisms and attempted to produce useful substances using these enzymes. (a) Enzymatic syntheses of various bioactive glycopeptides were carried out using transglycosylation activity of microbial glycosidases. (b) Effect of glycosylation on the function of protein was investigated using microbial glycosidases.
2. We found that aminopeptidases A, B and N, and dipeptidase D with broad substrate specificity are the four cysteinylglycinases of Escherichia coli K-12. Enzymatic characteristics of purified aminopeptidase B were determined.
3. We identified that the catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase is the Thrresidue at the N-terminal of the small subunit by a new novel affinity labeling agent.
4. We found that Gpr1, a putative G-protein coupled receptor, regulated glucose dependent cellular cAMP level in yeast Saccharomyces cerevisiae. Functional analysis revealed that pseudohyphal development observed under high glucose and low nitrogen condition was reduced in the GPR1 gene disruptant. We also clarified that the reduced transcriptional level of FLO11 gene which encodes cell surface protein involved in cell adhesion caused pseudohyphal defect in the GPR1 disruptant.

  • Research Products

    (58 results)

All Other

All Publications (58 results)

  • [Publications] 熊谷英彦: "チロシンフェノールリアーゼによるL-DOPAの酵素的合成法の開発と実用化"生物工学会誌. 76(10). 420-422 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Chemo-enzymatic synthesis of a novel glycopeptide using a microbial endoglycosidase."Carbohyd.Res.. 305. 415-422 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Chemo-enzymatic synthesis of calcitonin derivatives containing N-linked oligosaccharides."Bioorg.Med.Chem.Lett.. 8. 1303-1306 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Effect of carbohydrate chain on surface net charge and hydrophobicity of glycoproteins."Biosci.Biotech.Biochem.. 62(11). 2171-2176 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Characterization of Bacillus sp.endo-β-N-acetylglucosaminidase and its application to deglycosylation of hen ovomucoid."Biotechnol.Appl.Biochem.. 28(3). 235-242 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本憲二,熊谷英彦: "コリネ型細菌が生産する酵素の特異的な機能の利用"バイオサイエンスとインダストリー. 56(5). 313-315 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本憲二: "微生物のエンドグリコシダーゼの活性解析とその応用"蛋白質核酸酵素. 43(7). 866-874 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 玉置尚徳: "出芽酵母おける栄養源のシグナル伝達と2つのGタンパク質"化学と生物. 36(9). 589-590 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hisanori Tamaki et al.: "Gpr1p, a putative G-protein coupled receptor, regulates glucose dependent cellular cAMP level in yeast Saccharomyces cerevisiae."Biochem.Biophys.Res.Commun. 252(1). 29-33 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 熊谷英彦 ら: "ビフィズス菌由来の固定化酵素によるビフィズス因子の合成"ビタミン. 7. 403-405 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Transglycosylation activity of the endo-β-1,4-glucanase from Aspergillus niger IFO31125 and its application."J.Biosci.Bioeng.. 87(5). 576-580 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Glutathione metabolism in Escherichia coli."J.Mol.Catal.B. 6(3). 175-184 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Transcriptional regulation of tyrosine phenol-lyase gene mediated through TyrR and cAMP receptor protein."Biosci.Biotechnol.Biochem.. 63(10). 1823-1827 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hisanori Tamaki et al.: "Expression of two glutathione S-transferase genes in the yeast Issatchenkia orientalis is induced by o-dinitrobenzene during cell growth arrest."J.Bacteriol.. 181(9). 2958-2962 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 玉置尚徳: "出芽酵母において見つかったグルコース受容体"バイオサイエンスとインダストリー. 57(3). 179-180 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hidehiko Kumagai: "Microbial Production of Amino Acids in Japan"Adv.Biochem.Eng./Biotech.. 69. 72-85 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Purification, cloning, and 3-dimensional structure prediction of Micrococcus luteus FAD-containing tyramine oxidase."Biochem.Biophys.Res.Commun.. 268(2). 293-297 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Cloning and random mutagenesis of the Erwinia herbicola tyrR gene for high-level expression of tyrosine phenol-lyase."Appl.Environ.Microbiol.. 66(11). 4764-4771 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Characterization of endo-α-N-acetylgalactosaminidase from Bacillus sp. and syntheses of neo-oligosaccharides using its transglycosylation activity."Arch.Biochem.Biophys.. 373(2). 394-400 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "High-yield production and characterization of α-galactosidase from Bifidobacterium breve grown on raffinose."Biotechnol.Lett.. 22. 747-751 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Trypsin inhibitory activity of bovine fetuin de-O-glycosylated by endo-α-N-acetylgalactosaminidase."Biosci.Biotech.Biochem.. 64(10). 2266-2268 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kenji Yamamoto et al.: "Molecular cloning of cDNA encoding α-N-acetylgalactosaminidase from Acremonium sp.and its expression in yeast."Arch.Biochem.Biophys.. 384(2). 305-310 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid : N-terminal Thr-391 in small subunit is the nucleophile."Biochemistry. 39(26). 7764-7771 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hisanori Tamaki et al.: "GPR1 regulates filamentous growth through FLO11 in yeast Saccharomyces cerevisiae."Biochem.Biophys.Res.Commun.. 267(1). 164-168 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 玉置尚徳: "出芽酵母における栄養源認識と偽菌糸形成"生物工学会誌. 78(4). 139 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Aminopeptidases A, B and N, and dipeptidase D are the four cysteinylglycinases of Escherichia coli K-12."J.Bacteriol.. 183(4). 1489-1490 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 鈴木秀之,熊谷英彦: "γ-グルタミルトランスペプチダーゼ:Ntn-ハイドロラーゼスーパーファミリーの新しいメンバー"蛋白質核酸酵素. (印刷中). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hideyuki Suzuki et al.: "Purification and Characterization of Aminopeptidase B from Escherichia coli K-12."Biosci.Biotechnol.Biochem.,. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 山本憲二: "糖ペプチドの酵素合成 生理活性糖鎖研究法(生物化学実験法シリーズ)"学会出版センター. 168-175 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Yamamoto, K.Fujimori, K.Haneda, M.Mizuno, T.Inazu, and H.Kumagai.: "Chemo-enzymatic synthesis of a novel glycopeptide using a microbial endoglycosidase."Carbohyd.Res.. 305. 415-422 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Haneda, T.Inazu, M.Mizuno, R.Iguchi, K.Yamamoto, H.Kumagai, S.Aimoto, H.Suzuki, and T.Noda.: "Chemo-enzymatic synthesis of calcitonin derivatives containing N-linked oligosaccharides."Bioorg.Med.Chem.Lett.. 8. 1303-1306 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Akiba, K.Yamamoto, and H.Kumagai.: "Effect of carbohydrate chain on surface net charge and hydrophobicity of glycoproteins."Biosci.Biotech.Biochem.. 62(11). 2171-2176 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Yamamoto, T.Tanaka, K.Fujimori, C.-S.Kang, H.Ebihara, J.Kanamori, S.Kadowaki, T.Tochikura, and H.Kumagai.: "Characterization of Bacillus sp. endo-β-N-acetyl-glucosaminidase and its application to deglycosylation of hen ovomucoid."Biotechnol.Appl.Biochem.. 28(3). 235-242 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.-W.Yun, H.Tamaki, R.Nakayama, K.Yamamoto, and Hidehiko Kumagai.: "Gpr1p, a putative G-protein coupled receptor, regulates glucose dependent cellular cAMP level in yeast Saccharomyces cerevisiae."Biochem.Biophys.Res.Commun.. 252(1). 29-33 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Akiba, K.Yamamoto, and H.Kumagai.: "Transglycosylation activity of the endo-β-1,4-glucanase from Aspergillus niger IFO31125 and its application"J.Biosci.Bioeng.. 87(5). 576-580 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Katayama, H.Suzuki, K.Yamamoto, and H.Kumagai.: "Transcriptional regulation of tyrosine phenol-lyase gene mediated through TyrR and cAMP receptor protein."Biosci.Biotechnol.Biochem.. 63(10). 1823-1827 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Tamaki, K.Yamamoto, and H.Kumagai.: "Expression of two glutathione S-transferase genes in the yeast Issatchenkia orientalis is inducedby o-dinitro-benzene during cell growth arrest."J.Bacteriol.. 181(9). 2958-2962 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Ashida, K.Yamamoto, T.Murata, T.Usui, and H.Kumagai.: "Characterization of endo-α-N-acetylgalactosaminidase from Bacillus sp. and syntheses of neo-oligosaccharides using its transglycosylation activity."Arch.Biochem.Biophys.. 373(2). 394-400 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Xiao, K.Tanaka, X.M.Qian, K.Yamamoto, and H.Kumagai.: "High-yield production and characterization of α-galactosidase from Bifidobacterium brave grown on raffinose."Biotechnol.Lett.. 22. 747-751 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Ashida, K.Yamamoto, and H.Kumagai.: "Trypsin inhibitory activity of bovine fetuin de-O-glycosylated by endo-α-N-acetylgalactosaminidase."Biosci.Biotech.Biochem.. 64(10). 2266-2268 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Ashida, H.Tamaki, T.Fujimoto, K.Yamamoto, and H.Kumagai.: "Molecular cloning of cDNA encoding α-N-acetyl-galactosaminidase from Acremonium sp. and its expression in yeast."Arch.Biochem.Biophys.. 384(2). 305-310 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.H.Roh, J.Wouters, E.Depiereux, H.Yukawa, M.Inui, H.Minami, H.Suzuki, and H.Kumagai.: "Purification, cloning, and 3-dimensional structure prediction of Micrococcus luteus FAD-containing tyramine oxidase."Biochem.Biophys.Res.Commun.. 268(2). 293-297 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Katayama, H.Suzuki, T.Koyanagi, and H.Kumagai.: "Cloning and random mutagenesis of the Erwinia herbicola tyrR gene for high-level expression of tyrosine phenol-lyase."Appl.Environ.Microbiol.. 66(11). 4764-4771 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Inoue, J.Hiratake, H.Suzuki, H.Kumagai, and K.Sakata.: "Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid : N-terminal Thr-391 in small subunit is the nucleophile."Biochemistry. 39(26). 7764-7771 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Tamaki, T.Miwa, M.Shinozaki, M.Saito, C.-W.Yun, K.Yamamoto, and H.Kumagai.: "GPR1 regulates filamentous growth through FL011 in yeast Saccharomyces cerevisiae."Biochem.Biophys.Res.Commun.. 267(1). 164-168 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Suzuki, S.Kamatani, E.-S.Kim, and H.Kumagai.: "Aminopeptidases A, B and N, and dipeptidase D are the four cysteinylglycinases of Escherichia coli K-12."J.Bacteriol.. 183(4). 1489-1490 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Suzuki, S.Kamatani, and H.Kumagai.: "Purification and Characterization of Aminopeptidase B from Escherichia coli K-12."Biosci.Biotechnol.Biochem. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Kumagai: "Development and application of enzymatic production of L-DOPA by tyrosine phenol-lyase."Seibutsukougaku-kaishi. 76(10). 420-422 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Yamamoto, and H,Kumagai.: "Application of specific functions of enzymes produced from coryne-form bacteria."Bioscience and Industry. 56(5). 313-315 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Yamamoto.: "Microbial endoglycosidases and their application."Protein, Nuleic Acid and Enzyme. 43(7). 866-874 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Tamaki: "Role of two different G-proteins in nutrition signal transduction of budding yeast."Kagaku to Seibutsu. 36(9). 589-590 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Suzuki, W.Hashimoto, and H.Kumagai.: "Glutathione metabolism in Escherichia coli."J.Mol.Catal.B. 6(3). 175-184 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N,Nunoura, and H.Kumagai.: "Production of Bifidus-factor by immobilized enzyme from Bifidobacterium"Vitamin. 7. 403-405 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Tamaki: "Glucose receptor from budding yeast."Bioscience and Industry. 57(3). 179-180 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Tamaki: "Role of two different G-proteins in nutrition signal transduction of budding yeast."Seibutsukougaku-kaishi. 78(4). 139 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Kumagai.: "Microbial production of amino acids in Japan."Adv.Biochem.Eng./Biotcch.. 69. 72-85 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Suzuki, and H.Kumagai.: "γ-Glutamyltranspeptidase : A new member of Ntn hydrolase superfamily."Protein, Nuleic Acid and Enzyme. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Yamamoto.: "Enzymatic production of glycopeptide."Protocols in bioactive sugar chains. Gakkai-shuppann Center.. 168-175 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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