<p>Traditional Chinese medicines (TCM) represent a valuable resource for drug discovery, with numerous bioactive components identified with diverse pharmacological activities. However, the scarcity of these compounds, particularly those derived from plants, presents significant challenges for their applications. Here, we develop a modular and concise chemoenzymatic platform that features a streamlined enzymatic cascade coupled with a simple chemical module to facilitate the highly cost-effective and sustainable manufacture of bioactive components from Coptidis Rhizoma (Huanglian). After enzyme screening and engineering, as well as the implementation of a “plug-and-play” strategy, we efficiently produce various protoberberine alkaloids, including berberine (44% yield), demethyleneberberine (48% yield), jatrorrhizine (39% yield), berberrubine (31% yield), columbamine (42% yield), and palmatine (44% yield), exhibiting high robustness and industrial potential of this designed chemoenzymatic platform. This approach not only addresses challenges associated with heterologous production in engineered cells and low regioselectivity in chemical synthesis, but also establishes a paradigm for manufacturing other bioactive components derived from TCM, further facilitating the modernization of TCM.</p>