Phylogenetic analysis of Bacillus P450 monooxygenases and evaluation of their activity towards steroids

Toshiki Furuya, Daisuke Shibata, Kuniki Kino

    Research output: Contribution to journalArticle

    16 Citations (Scopus)

    Abstract

    Cytochrome P450 (P450) open reading frames (ORFs) identified in genome sequences of Bacillus species are potential resources for new oxidation biocatalysts. Phylogenetic analysis of 29 Bacillus P450 ORFs revealed that the P450s consist of a limited number of P450 families, CYP102, CYP106, CYP107, CYP109, CYP134, CYP152, and CYP197. Previously, we identified the catalytic activities of three P450s of Bacillus subtilis towards steroids by rapid substrate screening using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). Here, we further applied this method to evaluate the activity of Bacillus cereus P450s towards steroids. Five P450 genes were cloned from B. cereus ATCC 10987 based on its genomic sequence and were expressed in Escherichia coli. These P450s were reacted with a mixture of 30 compounds that mainly included steroids, and the reaction mixtures were analyzed using FT-ICR/MS. We found that BCE_2659 (CYP106) catalyzed the monooxygenation of methyltestosterone, progesterone, 11-ketoprogesterone, medroxyprogesterone acetate, and chlormadinone acetate. BCE_2654 (CYP107) monooxygenated testosterone enanthate, and BCE_3250 (CYP109) monooxygenated testosterone and compactin. Based on the phylogenetic relationship and the known substrate specificities including ones identified in this study, we discuss the catalytic potential of Bacillus P450s towards steroids.

    Original languageEnglish
    Pages (from-to)906-912
    Number of pages7
    JournalSteroids
    Volume74
    Issue number12
    DOIs
    Publication statusPublished - 2009 Nov 4

    Fingerprint

    Bacilli
    Mixed Function Oxygenases
    Bacillus
    Steroids
    Bacillus cereus
    Cyclotrons
    Cyclotron resonance
    Fourier Analysis
    Open Reading Frames
    Mass spectrometry
    Mass Spectrometry
    Fourier transforms
    Genes
    Chlormadinone Acetate
    Ions
    Methyltestosterone
    Medroxyprogesterone Acetate
    Substrates
    Substrate Specificity
    Bacillus subtilis

    Keywords

    • Bacillus
    • Biotransformation
    • Cytochrome P450
    • Mass spectrometry
    • Monooxygenase

    ASJC Scopus subject areas

    • Biochemistry
    • Clinical Biochemistry
    • Endocrinology
    • Molecular Biology
    • Organic Chemistry
    • Pharmacology

    Cite this

    Phylogenetic analysis of Bacillus P450 monooxygenases and evaluation of their activity towards steroids. / Furuya, Toshiki; Shibata, Daisuke; Kino, Kuniki.

    In: Steroids, Vol. 74, No. 12, 04.11.2009, p. 906-912.

    Research output: Contribution to journalArticle

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