Expression, purification and crystallization of a plant polyketide cyclase from Cannabis sativa

Xinmei Yang, Takashi Matsui, Takahiro Mori, Futoshi Taura, Hiroshi Noguchi, Ikuro Abe, Hiroyuki Morita

    Research output: Contribution to journalArticlepeer-review

    2 Citations (Scopus)

    Abstract

    Plant polyketides are a structurally diverse family of natural products. In the biosynthesis of plant polyketides, the construction of the carbocyclic scaffold is a key step in diversifying the polyketide structure. Olivetolic acid cyclase (OAC) from Cannabis sativa L. is the only known plant polyketide cyclase that catalyzes the C2-C7 intramolecular aldol cyclization of linear pentyl tetra-β-ketide-CoA to generate olivetolic acid in the biosynthesis of cannabinoids. The enzyme is also thought to belong to the dimeric +β barrel (DABB) protein family. However, because of a lack of functional analysis of other plant DABB proteins and low sequence identity with the functionally distinct bacterial DABB proteins, the catalytic mechanism of OAC has remained unclear. To clarify the intimate catalytic mechanism of OAC, the enzyme was overexpressed in Escherichia coli and crystallized using the vapour-diffusion method. The crystals diffracted X-rays to 1.40Å resolution and belonged to space group P3121 or P3221, with unit-cell parameters a = b = 47.3, c = 176.0Å. Further crystallographic analysis will provide valuable insights into the structure-function relationship and catalytic mechanism of OAC.

    Original languageEnglish
    Pages (from-to)1470-1474
    Number of pages5
    JournalActa Crystallographica Section:F Structural Biology Communications
    Volume71
    DOIs
    Publication statusPublished - 2015

    Keywords

    • Cannabis sativa
    • cannabinoids
    • cyclase
    • olivetolic acid

    ASJC Scopus subject areas

    • Biophysics
    • Structural Biology
    • Biochemistry
    • Genetics
    • Condensed Matter Physics

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