Steady-State Kinetic Characterization of Evolved Biphenyl Dioxygenase, Which Acquired Novel Degradation Ability for Benzene and Toluene

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Biphenyl dioxygenase (Bph Dox) catalyzes initial oxygenation in the bacterial biphenyl degradation pathway. Bph Dox in <I>Pseudomonas pseudoalcaligenes</I> KF707 is a Rieske type three-component enzyme in which a large subunit (encoded by the <I>bphA1</I> gene) plays an important role in the substrate specificity of Bph Dox. Steady-state kinetic assays using purified enzyme components demonstrated that KF707 Bph Dox had a <I>k</I><SUB>cat</SUB>⁄<I>K</I><SUB>m</SUB> of 33.1×10<SUP>3</SUP> (<small>M</small><SUP>−1</SUP> s<SUP>−1</SUP>) for biphenyl. Evolved 1072 Bph Dox generated by the process of DNA shuffling (Suenaga, H. <I>et al.</I>, <I>J. Bacteriol.</I>, <B>184</B>, 3682–3688 (2002)) exhibited enhanced degradation activity not only for biphenyl (<I>k</I><SUB>cat</SUB>⁄<I>K</I><SUB>m</SUB> of 62.2×10<SUP>3</SUP> [<small>M</small><SUP>−1</SUP> s<SUP>−1</SUP>]) but also for benzene and toluene, compounds that are rarely attacked by KF707 Bph Dox. These results suggest that evolved 1072 Bph Dox acquires higher affinities and catalytic efficiencies for various substrates than the original KF707 enzyme.

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  • Bioscience, biotechnology, and biochemistry  

    Bioscience, biotechnology, and biochemistry 70(4), 1021-1025, 2006-04-23 

    Japan Society for Bioscience, Biotechnology, and Agrochemistry

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  • NII論文ID(NAID)
    10018532876
  • NII書誌ID(NCID)
    AA10824164
  • 本文言語コード
    ENG
  • 資料種別
    NOT
  • ISSN
    09168451
  • NDL 記事登録ID
    7899991
  • 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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