Structural Basis for β-Carboline Alkaloid Production by the Microbial Homodimeric Enzyme McbB

Takahiro Mori, Shotaro Hoshino, Shusaku Sahashi, Toshiyuki Wakimoto, Takashi Matsui, Hiroyuki Morita, Ikuro Abe

    Research output: Contribution to journalArticlepeer-review

    28 Citations (Scopus)

    Abstract

    Summary The β-carboline (βC) alkaloids occur throughout nature and exhibit diverse biological activities. In contrast to βC alkaloid synthesis in plants, the biosynthesis in microorganisms remains poorly understood. The recently reported McbB from Marinactinospora thermotolerans is a novel enzyme proposed to catalyze the Pictet-Spengler (PS) reaction of L-tryptophan and oxaloacetaldehyde to produce the βC scaffold of marinacarbolines. In this study, we solved the crystal structure of McbB complexed with L-tryptophan at 2.48 Å resolution, which revealed the novel protein folding of McbB and the totally different structure from those of other PS condensation catalyzing enzymes, such as strictosidine synthase and norcoclaurine synthase from plants. Structural analysis and site-directed mutagenesis confirmed that the previously proposed catalytic Glu97 at the active-site center functions as an acid and base catalyst. Remarkably, the structure-based mutants R72A and H87A, with expanded active-site cavities, newly accepted bulky phenylglyoxal as the aldehyde substrate, to produce 1-benzoyl-3-carboxy-β-carboline.

    Original languageEnglish
    Article number3075
    Pages (from-to)898-906
    Number of pages9
    JournalChemistry and Biology
    Volume22
    Issue number7
    DOIs
    Publication statusPublished - 2015 Jul 24

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Medicine
    • Molecular Biology
    • Pharmacology
    • Drug Discovery
    • Clinical Biochemistry

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