Cloning and Expression of Pseudomonas taetrolens Y-30 Gene Encoding Glutamine Synthetase : An Enzyme Available for Theanine Production by Coupled Fermentation with Energy Transfer

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Glutamine synthetase (GS) of <I>Pseudomonas taetrolens</I> Y-30 can form theanine from glutamic acid and ethylamine in a mixture where yeast fermentation of sugar is coupled for ATP regeneration (coupled fermentation with energy transfer). From a genomic DNA library of <I>P. taetrolens</I> Y-30, a clone containing 6 kbp insertional DNA fragment was selected by the PCR screening technique with specific oligonucleotide primers for the GS gene. The fragment had an open reading frame of the GS gene encoding a protein of 468 amino acids (molecular mass, 52 kDa). The deduced amino acid sequence showed a significant homology with that of <I>P. syringae pv. tomato</I> GS (97%), and all the amino acid residues were fully conserved, which concern with catalytic activity in other bacterial GS. A tyrosine residue for adenylylation of GS was also found, and <I>in vivo</I> adenylylation was confirmed in <I>P. taetrolens</I> Y-30. The isolated GS gene was ligated into an expression vector (pET21a), and expressed in <I>Escherichia coli</I> AD494 (DE3). The enzyme productivity in the expression system was 30-fold higher than that in <I>P. taetrolens</I> Y-30. Recombinant GS had the same properties as those of unnadenylylated intrinsic GS, and formed theanine in the mixture of coupled fermentation with energy transfer.

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  • Bioscience, biotechnology, and biochemistry  

    Bioscience, biotechnology, and biochemistry 70(2), 500-507, 2006-02-23 

    Japan Society for Bioscience, Biotechnology, and Agrochemistry

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各種コード

  • NII論文ID(NAID)
    10018534572
  • NII書誌ID(NCID)
    AA10824164
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    09168451
  • NDL 記事登録ID
    7833576
  • NDL 雑誌分類
    ZR7(科学技術--農林水産--農産) // ZR2(科学技術--生物学--生化学) // ZP1(科学技術--化学・化学工業)
  • NDL 請求記号
    Z53-G223
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
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