2003 Fiscal Year Final Research Report Summary
Molecular and cellular research on aromatic amino acids production, by microorganisms.
Project/Area Number |
14360056
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Research Category |
Grant-in-Aid for Scientific Research (B)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
応用微生物学・応用生物化学
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Research Institution | KYOTO 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)
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Project Period (FY) |
2002 – 2003
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Keywords | Tyrosine phenol lyase / TyrR / DOPA / Erwinia herbicola / tutB / LIVI / LS system / transporter / aromatic amino acids |
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.
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Research Products
(6 results)