Biochemical Characterization of Ornithine Decarboxylases from Solanaceae Plants Producing Tropane Alkaloids

Ornithine decarboxylase (ODC) is the rate-limiting enzyme in the biosynthesis of polyamines and plant alkaloids, including medicinal tropane alkaloids (TAs). Due to its key role, ODC has been utilized as an effective molecular tool in metabolic engineering. However, to date, only a limited number of plant ODCs have been characterized. Among the reported ODCs, Erythroxylum coca ODC (EcODC) exclusively has ODC activity, while Nicotiana glutinosa ODC (NgODC) exhibits dual ODC and lysine decarboxylase (LDC) activities. The potential LDC activity of ODCs from TA-producing plants remains unknown. Here, we characterized AlODC and DsODC from Anisodus luridus and Datura stramonium, along with two previously reported ODCs from Atropa belladonna (AbODC) and Hyoscyamus niger (HnODC), in Escherichia coli to investigate their enzyme kinetics and substrate specificity. Enzymatic assays revealed that both AlODC and DsODC catalyzed the conversion of ornithine to putrescine, confirming their ODC activity, with AlODC exhibiting a higher catalytic efficiency, comparable to established ODCs. Furthermore, all four ODCs also displayed LDC activity, albeit at significantly lower efficiency (<1% of ODC activity). This study provides a comprehensive analysis of the enzyme kinetics of ODCs from TA-producing plants, identifying promising candidate genes for metabolic engineering for the biomanufacturing of putrescine-derived alkaloids. Moreover, this is the first report of LDC activity in ODCs from Solanaceae TA-producing plants, shedding light on the evolutionary relationship between ODC and LDC.