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Studies on structure and function of amine and amino acid related enzymes from microbiology.

Research Project

Project/Area Number 06453168
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 InstitutionKYOTO UNIVERSITY

Principal Investigator

KUMAGAI Hidehiko  Kyoto University, Faculty of Agriculture, Professor, 農学部, 教授 (70027192)

Co-Investigator(Kenkyū-buntansha) TAMAKI Hisanori  Kyoto University, Faculty of Agriculture, Instructor, 農学部, 助手 (20212045)
SUZUKI Hideyuki  Kyoto University, Faculty of Agriculture, Instructor, 農学部, 助手 (10202136)
YAMAMOTO Kenji  Kyoto University, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (70109049)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1995: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥4,000,000 (Direct Cost: ¥4,000,000)
Keywordsgamma-glutamyltranspeptidase / monoamine oxidase / tyrosine phenol-lyase / Bifidobacterium / beta-glucosidase / amine oxidase / glutathione S-transferase / モノアミン酸化酵素 / アミン酸化酵素 / グルタチオンS-トランスフェラーゼ / X線結晶解析 / γ-グルタミルトランスフェラーゼ
Research Abstract

1. (1) Lys-257 of tyrosine phenol-lyase was mutated into Ala. And the strain over expresses the mutant enzyme was constructed using E.coli and T7 phage. (2) Transcription initiation site and regulation sites of tyrosine phenol-lyase gene was analyzed.
2. (1) Structure of the main chain of gamma-glutamyltranspeptidase was determined. (2) Ser-33 of gamma-glutamyltranspeptidase was mutated into Cys. Pb^<2+> derivatives of the enzyme was analyzed. Mechanism of low temperature induction of the enzyme was analyzed. The induction was regulated by transcriptional regulation.
3. (1) Monoamine oxidase of E.coli was induced by Cu^<2+> ion and the induction was regulated by transcriptional regulation. (2) The enzyme was induced by tyramine and also regulated by catabolite repression. (3) Over-expression of the regulator protein results in production of yellow enzyme. The yellow enzyme was purified and characterized. Its crystal structure was also studied.
4.Copper-binding His residues and active center amino acid residues of amine oxidase of fungi were mutagenized.
5.Isoenzyme gene of glutathione S-transferase was cloned from yeast. Over-expressed enzyme was purified and characterized.
6.beta-Glucosidase gene of bifidobacterium was cloned in E.coli. Over-expressed enzyme was purified, characterized, and crystallized.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Mitsuo Yamashita: "maoB, a gene that encodes a positive regulator of the monoamine oxidase gene (maoA) in Escherichia coli." Journal of Bacteriology. 178 (10). 2941-2947 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hiroaki Sakai: "A nobel protein structure : γ-glutamyltranspeptidase from E. coli K-12." Journal of Biochemistry. 120 (1). 26-28 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 熊谷英彦: "L-ドーパの酵素的合成法の開発と実用化" バイオサイエンスとインダストリー. 54 (1). 11-15 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ivo Frebort: "Two distinct quinoprotein amine oxidases are induced by n-butylamine in the mycelia of Aspergillus niger AKU 3302." European Journal of Biochemistry. 237 (1). 255-265 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Naoki Nunoura: "Purification and characterization of β-D-glucosidase (β-D-fucosidase) from Bifidobacterium breveclb acclimated to cellobiose." Bioscience Biotechnology and Biochemistry. 60 (2). 188-193 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Wataru Hahimoto: "Low temperature inducible γ-glutamyltranspeptidase of Escherichia coli K-12." Bioscience Biotechnology and Biochemistry. 61 (1). 34-39 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Yamashita: "maoB,a gene that encodes a positive regulator of the monoamine oxidase gene (maoA) in Escherichia coli" J.Bacteriol.178. 2941-2947 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Sakai: "A nobel protein structure : gamma-glutamyltranspeptidase from E.coli K-12" J.Biochem.120. 26-28 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Kumagai: "Development of L-DOPA production by enzymatic synthesis" Bioscience and Technology. 54. 11-15 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] I.Frebort: "Two distinct quinoprotein amine oxidase by n-butylamine in the mycelia of Aspergillus niger AKU 3302" Eur.J.Biochem.60. 188-193 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] N.Nunoura: "Purification and characterization of beta-D-glucosidase (beta-D-fucosidase) from Bifidobacteriumbreve clb acclimated to cellobiose" Biosci. Biotechnol. Biochem.60. 188-193 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] W.Hashimoto: "Low temperature inducible gamma-glutamyltranspeptidase of Escherichia coli K-12" Biosci. Biotechnol. Biochem.61. 34-39 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Mitsuo Yamashita: "maoB,a gene that encodes a positive regulator of the monoamine oxidase gene (maoA) in Escherichia coli." Journal of Bacteriology. 178(10). 2941-2947 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroaki Sakai: "A nobel protein structure : γ-glutamyltranspeptidase from E.coli K-12." Journal of Biochemistry. 120(1). 26-28 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 熊谷英彦: "L-ドーパの酵素的合成法の開発と実用化" バイオサイエンスとインダストリー. 54(1). 11-15 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ivo Frebort: "Two distinct quinoprotein amine oxidases are induced by n-butylamine in the mycelia of Aspergillus niger AKU 3320." European Journal of Biochemistry. 237(1). 255-265 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Naoki Nunoura: "Purification and characterization of β-D-glucosidase (β-D-tucosidase)from Bitidobacterium breve clb acclimated to cellobiose." Bioscience Biotechnology and Biochemistry. 60(2). 188-193 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wataru Hahimoto: "Low temperature inducible γ-glutamyltranspeptidase of Escherichia coli K-12." Bioscience Biotechnology and Biochemistry. 61(1). 34-39 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ivo Frebort, et al: "Two distinct quinoprotein amine oxidases are induced by n-butylamine in mycelia of Aspergillus niger AKU 3302." Eur. J. Biochem.,. (in press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hideyuki Suzuki, et al: "Transcriptional regulation of tyrosine phenol lyase gene of Erwinia herbicola AJ2985." Biosci. Biotech. Biochem.59. 2339-2341 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Jung-Hyeob Rob,et al: "Escherichia coli K-12 copper-containing monoamine oxidase:" Biochem. Biophys. Res. Commun.212. 1107-1114 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Suzuki.et al: "Escherichia coli k-12 can utilize an exogenous γ-glutamyl peptide as an amino acid source,for which γ-glutamyltranspeptidase is essential." J.Bacteriol.175. 6038-6040 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] H.Kumagai,et al: "Crystallization and preliminary X-ray analysis of γ-glutamyltranspeptidase from Escherichia coliK-12." J.Mol.Biol.234. 1259-1262 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Jung Hyeob Roh,et al: "Crystallization and preliminary X-ray analysis of copper amine oxidase from Escherichia coliK-12." J.Mol.Biol.238. 635-637 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Jung Hyeob Roh,et al: "Purification,characterization,and crystallization of monoamine oxidase from Eschericha coliK-12." Biosci.Biotech.Biochem.58. 1652-1656 (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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