Tropinone synthesis via an atypical polyketide synthase

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Tropinone synthesis via an atypical polyketide synthase

Tropinone is the first intermediate in the biosynthesis of the pharmacologically important tropane alkaloids that possesses the 8-azabicyclo[3.2.1]octane core bicyclic structure that defines this alkaloid class. Chemical synthesis of tropinone was achieved in 1901 but the mechanism of tropinone biosynthesis has remained elusive. In this study, we identify a root-expressed type III polyketide synthase from Atropa belladonna (AbPYKS) that catalyzes the formation of 4-(1-methyl-2-pyrrolidinyl)-3-oxobutanoic acid. This catalysis proceeds through a non-canonical mechanism that directly utilizes an unconjugated N-methyl-Δ1-pyrrolinium cation as the starter substrate for two rounds of malonyl-Coenzyme A mediated decarboxylative condensation. Subsequent formation of tropinone from 4-(1-methyl-2-pyrrolidinyl)-3-oxobutanoic acid is achieved through cytochrome P450-mediated catalysis by AbCYP82M3. Silencing of AbPYKS and AbCYP82M3 reduces tropane levels in A. belladonna. This study reveals the mechanism of tropinone biosynthesis, explains the in planta co-occurrence of pyrrolidines and tropanes, and demonstrates the feasibility of tropane engineering in a non-tropane producing plant.ISOPROPYLBENZYLAMINE

For millennia, plants have been utilized by humans as medicines, and their associated bioactivity is now attributed to chemically diverse specialized metabolites that often display lineage-specific distribution1,2,3. Identifying these bioactive components, determining their structures, resolving their underlying biosynthetic pathways, and attempting to metabolically engineer plants and microbes to boost yields or enhance the production of specific metabolites has occupied scientists for the last 150 years4,5,6,7,8,9. However, the biosynthesis of many plant-derived medicines remains unresolved.

Tropane alkaloids are a diverse class of ~200 plant-specialized metabolites that display selective distribution across the plant kingdom. They include the pharmaceuticals hyoscyamine (17) and scopolamine (18) that are synthesized by select genera of the Solanaceae family and the narcotic cocaine that is synthesized in the Erythroxylaceae10. Tropane alkaloids are defined by a characteristic 8-azabicyclo[3.2.1]octane core skeleton that is comprised of a cycloheptane ring spanned by a nitrogen bridge. Chemical diversity is manifest through varied decorations of the cycloheptane ring, including hydroxylations and acylations10. The biosynthesis of tropane alkaloids is not fully understood and in particular, the enzymes that catalyze the formation of the bicyclic ring common to all tropanes remains elusive.