Engineering an Artificial Taxol Biosynthetic Pathway from Baccatin III in Yeast

Abstract

Paclitaxel (Taxol) is among the most successful natural anticancer agents, yet its manufacturing still relies on expensive semisynthetic methods carried out under harsh alkaline conditions. Deciphering the entire Taxol biosynthetic pathway has been a longstanding challenge for over 50 years because of its complex regulatory network, but it has only recently been understood. Differences between plant and microbial systems render current knowledge insufficient for scalable biotechnological production, necessitating systematic metabolic engineering to overcome multiple bottlenecks. Artificial pathways can compensate for an incomplete understanding of native biosynthesis and offer an alternative strategy to alleviate supply constraints. Here, we report the construction of an artificial pathway in yeast that uses (2R,3S)-3-phenylisoserine as the substrate and requires only three enzymatic steps to convert baccatin III to Taxol, demonstrating the broad substrate scope of β-phenylalanoyl-CoA ligases (PCLs). Structure–activity analysis of Baccatin III:3-amino-3-phenylpropanoyltransferase (BAPT) is further employed to enhance its catalytic efficiency toward non-natural substrates. This work offers a practical strategy for the first step of sustainable production of Taxol using synthetic biology.