N-Debenzoyl-2'-deoxypaclitaxel:N-benzoyltransferase (NDTBT) isolated from Taxus plants is the last aroyltransferase on the paclitaxel biosynthesis pathway that catalyzes the formation of a benzamide functional group from co-substrates benzoyl CoA and N-debenzoyl-2'-deoxypaclitaxel. To determine substrate specificity, heterologously expressed NDTBT was purified for in vitro assays with three semisynthetically-derived N-debenzoylpaclitaxel substrates and several aroyl CoA donors (benzoyl; ortho-, meta-, and para-substituted benzoyls; various heterole carbonyls; alkanoyls; and butenoyl), which were obtained from commercial sources or synthesized via a mixed anhydride method. The biocatalytically assembled paclitaxel analogues had catalytic efficiencies (Vmax/KM) ranging from 0.15 to 1.74 nmol min-1 mM-1 when N-debenzoyl-2'-deoxypaclitaxel was assayed. When incubated with either N-debenzoylpaclitaxel or 10-deacetyl-N-debenzoylpaclitaxel, the relative velocity (vrel) values were compared ranging from ~1% to 200% for the aroyl CoAs compared to benzoyl CoA. The broad specificity of NDTBT's biocatalytic N-aroylation/alkanoylation could potentially facilitate large-scale semisynthetic processes that transfer unnatural acyl groups to similar diterpenoid compounds.

<i>N</i>-Benzoyltransferase has broad aroyl CoA specificity in vitro with analogs of <i>N</i>-debenzoylpaclitaxel

239th National Meeting (2010)

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N-Benzoyltransferase has broad aroyl CoA specificity in vitro with analogs of N-debenzoylpaclitaxel