Chemoenzymatic Synthesis and Protein Engineering Enable Efficient, Scalable Production of Teleocidin Derivatives

Abstract

Monoterpenoid indole alkaloids (MIAs), a class of bioactive natural products, are highly valued in drug development for their unique pharmacological activities. Teleocidins, known for activating protein kinase C (PKC), are particularly promising but challenging to synthesize due to their structural complexity. Traditional methods often rely on heavy metals and yield low amounts, while biosynthetic approaches face efficiency issues. Our study developed an efficient chemoenzymatic route to produce 13 teleocidin B compounds and derivatives at scale. To overcome enzymatic reaction bottlenecks, we engineered the critical enzyme TleB by fusing a reductase module to create a self‐sufficient P450 system, boosting indolactam V production to 868.8 mg L−1. Additionally, we established a dual‐cell factory co‐expressing engineered hMAT2A‐TleD and TleB/TleC enzymes, enabling the first fully enzymatic synthesis of teleocidin B isomers with a total yield of 714.7 mg L−1. Chemical modifications further expanded the library with five novel indolactam V and two teleocidin A1 derivatives. Fermentation confirmed the recombinant Escherichia coli system's scalability, producing 430 mg indolactam V, 170 mg teleocidin A1, and 300 mg teleocidin B isomers. This work not only establishes a sustainable platform for teleocidin synthesis but also addresses efficiency and scalability challenges in complex natural product synthesis, paving the way for practical applications of bioactive compounds.