Discovery of 2-naphthoic acid monooxygenases by genome mining and their use as biocatalysts

Toshiki Furuya, Kuniki Kino

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    The large pool of cytochrome P450 (P450) open-reading frames identified in genome sequences has attracted much attention as a resource for new oxidation biocatalysts. P450 genes were cloned from genome-sequenced bacteria and coexpressed with putidaredoxin and its reductase genes to provide the redox partners of P450 in Escherichia coli. Wholecell assays were performed with 2-naphthoic acid as a substrate. Hydroxylated naphthoic acid products were rapidly detected with two reagents showing different colors in the presence of the products. Two P450s, CYP199A1 and CYP199A2, were found to hydroxylate the substrate to 7-and 8-hydroxy-2-naphthoic acids. The CYP199A1 whole-cell biocatalyst converted 1 mm 2-naphthoic acid to 0.27 mm 7-hydroxy-2-naphthoic acid and 0.53 mm 8-hydroxy-2-naphthoic acid. CYP199A2 exhibited similar regioselectivity to CYP199A1. Furthermore, we found that 8-hydroxy-2-naphthoic acid emits near-white fluorescence when exposed to UV light. These P450s will provide a facile and environmentally friendly synthetic approach to the hydroxynaphthoic acids.

    Original languageEnglish
    Pages (from-to)645-649
    Number of pages5
    JournalChemSusChem
    Volume2
    Issue number7
    DOIs
    Publication statusPublished - 2009 Jul

    Fingerprint

    Biocatalysts
    Mixed Function Oxygenases
    genome
    Genes
    Acids
    acid
    Enzymes
    Regioselectivity
    Substrates
    substrate
    Ultraviolet radiation
    Cytochrome P-450 Enzyme System
    Escherichia coli
    2-naphthoic acid
    gene
    Assays
    Bacteria
    Fluorescence
    Color
    cytochrome

    Keywords

    • Biocatalysis
    • Cytochromes
    • Enzymes
    • Hydroxylation
    • Oxidation

    ASJC Scopus subject areas

    • Energy(all)
    • Environmental Chemistry
    • Materials Science(all)
    • Chemical Engineering(all)

    Cite this

    Discovery of 2-naphthoic acid monooxygenases by genome mining and their use as biocatalysts. / Furuya, Toshiki; Kino, Kuniki.

    In: ChemSusChem, Vol. 2, No. 7, 07.2009, p. 645-649.

    Research output: Contribution to journalArticle

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