A dual self-regulatory platform for programmable biosynthesis of L-lysine-derived alkaloids

L-lysine-derived alkaloids represent a structurally diverse and pharmacologically significant class of nitrogen-containing natural products with broad utility as pharmaceutical agents and industrial platform chemicals. However, their microbial biosynthesis has been persistently hindered by precursor imbalance and the rigid regulatory architecture of native metabolic pathways. Here, we report a programmable biosynthetic platform for the de novo production of L-lysine-derived alkaloids directly from glucose in Corynebacterium glutamicum . Through systematic metabolic rewiring, we developed a high-flux cadaverine-producing chassis that achieves robust accumulation of the central nitrogenous precursor required for downstream alkaloid assembly. Leveraging this chassis, we designed a dual metabolite-responsive regulatory circuit that autonomously orchestrates malonate assimilation, malonyl-CoA supply, and alkaloid formation without external inducers. This self-regulating strategy enabled the production of pelletierine at 6.11 ± 0.06 g/L in fed-batch fermentation, representing the highest reported microbial titer to date. Extending this platform through selective short-chain dehydrogenase/reductase catalysis, we further achieved the stereoselective conversion of 1-piperideine and pelletierine into piperidine (8.60 ± 0.16 g/L) and sedridine (3.71 ± 0.19 g/L), respectively—constituting a heterologous microbial production of either compound. Collectively, this work establishes a versatile, programmable, and industrially tractable platform for L-lysine-derived alkaloid biosynthesis, providing a compelling foundation for the sustainable microbial production of diverse alkaloid intermediates and drug leads.