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

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Plant polyketides are a structurally diverse family of natural products. In the biosynthesis of plant polyketides, the construction of the carbocyclic scaffolds is a key step for 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 cannabinoid. The enzyme is also thought to belong to the dimeric α+β barrel (DABB) protein family. However, because of lack of the functional analysis of the other plant DABB proteins and low sequence identity with the functionally and structually characterized 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 in a vapour-diffusion method. The crystals diffracted X-rays to 1.40 Å resolutions and belonged to space group P3121 or P3221, with unit-cell parameters a = b = 47.3 Å, c = 176.0 Å. Further crystallographic analysis will provide valuable insights into the structure–function relationship and catalytic mechanism of OAC.

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Acta Crystallogr F Struct Biol Commun, 2015 Dec; 71(Pt 12): 1470-4

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