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Molecular and cellular research on aromatic amino acids production, by microorganisms.

Research Project

Project/Area Number 14360056
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 Grad.Sch.of Biostudies, Div.of Integrated Life Science, Professor, 生命科学研究科, 教授 (70027192)

Co-Investigator(Kenkyū-buntansha) SUZUKI Hideyuki  KYOTO UNIVERSITY Grad.Sch.of Biostudies, Div.of Integrated Life Science, Associate Professor, 生命科学研究科, 助教授 (10202136)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥17,200,000 (Direct Cost: ¥17,200,000)
Fiscal Year 2003: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥13,900,000 (Direct Cost: ¥13,900,000)
KeywordsTyrosine phenol lyase / TyrR / DOPA / Erwinia herbicola / tutB / LIVI / LS system / transporter / aromatic amino acids / TutB / LIV-I / DOPA / Ervinia herbicola / Erwinia harbicola / 転写調節
Research Abstract

In Escherichia coli, the active transport of phenylalanine is considered to be performed by two different systems, AroP and PheP However, a low. level of accumulation of phenylalanine was observed in an aromatic amino acid transporter-deficient E. coli strain (ΔaroP ΔpheP Δmtr Δtna ΔtyrP). The uptake of phenylalanine by this strain was significantly inhibited in the presence of branched-chain amino acids. Genetic analysis and transport studies revealed that the LIV-I/LS system, which is a branched-chain amino acid transporter consisting of two periplasmic binding proteins, the LIV-binding protein (LIV-I system) and LS-binding protein (LS system), and membrane components, LivHMGF, is involved in phenylalanine accumulation.in. E. coli. cells. The Km values for phenylalanine in the-LIV-I and LS systems were determined to be 19 and 30 μM, respectively. Competitive inhibition of phenylalanine uptake by isoleucine, leucine, and valine was observed for the LIV-I system and, surprisingly, also for the LS system, which has been assumed to be leucine specific on the basis of the results of binding studies with the purified LS-binding protein. We found that the LS system is capable of transporting isoleucine and valine with affinity comparable to that far leucine and that the LIV-I system is able to transport tyrosine with affinity lower than that seen with other substrates. The physiological importance of the LIV-I/LS system for phenylalanine accumulation was revealed in the growth of phenylalanine-auxotrophic E. coli strains under various conditions.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] T.Katayama et al.: "Functional analysis of the Erwinia herbicola tutB gene and its product."Journal of Bacteriology. 184(11). 3135-3141 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 片山高嶺 他: "L-ドーパ生産酵素チロシンフェノールリアーゼ(TPL)の発現調節機構の解明と転写調節因子の改変によるTPLの高発現"応用微生物学研究. 1(2). 130-137 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Koyanagi et al.: "Identification of the LIV-I/LS system as the third phenylalanine transporter in Escherichia coli K-12."Journal of Bacteriology. 186(2). 343-350 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Katayama, H.Suzuki, T.Koyanagi, H.Kumagai.: "Functional analysis of the Erwinia. herbicola tutB gene and its product."Journal of Bacteriology. 184(11). 3135-3141 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Katayama, T.Koyanagi, H.Suzuki, H.Kumagai.: "Expression of tyrosine phenol lyase which is used for L-DOPA production. The regulation of its expression and over-expression by its mutant regulators."Ouyou Biseibutsugaku Kennkyu. 1(2). 130-137 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.Koyanagi, T.Katayama, H.Suzuki, H.Kumagai.: "Identification of the LIV-I/LS system as the third phenylalanine transporter in Escherichia coli K-12."Journal of Bacteriology. 186(2). 343-350 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 片山高嶺 ほか: "L-ドーパ生産酵素チロシンフェノールリアーゼ(TPL)の発現 調節機構の解明と転写調節因子の改変によるTPLの高発現"応用微生物学研究. 1(2). 130-137 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Koyanagi et al.: "Identification of the LIV-I/LS system as the third phenylalanine transporter in Escherichia coli K-12"Journal of Bacteriology. 186(2). 343-350 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.katayama, H.Suzuki, T.Koyanagi, H.Kumagai: "Functional analysis of the Erwinia herbicola tutB gene and its product"Journal of Bacteriology. 184(11). 3135-3141 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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