Harnessing Gene Mining for Efficient (+)-Cedrol Synthesis in Yeast and Exploring its Anticancer Effects on Liver Cancer.
Plant-derived sesquiterpenes offer untapped opportunities for cancer therapeutics. Here, we strategy that unites gene mining, computational enzyme engineering, and metabolic rewiring to achieve record-level biosynthesis of (+)-cedrol. Screening terpene synthase homologues identified an ancestral enzyme with latent activity, which was rationally redesigned via structure-guided mutagenesis and molecular dynamics simulations to deliver >95% enantiomeric excess. By rationally rewiring the mevalonate pathway and optimizing fermentation processes in Saccharomyces cerevisiae, we achieved an unprecedented titre of 7109 mg/L-a 19,500-fold increase over baseline production levels. Pharmacological characterization revealed novel antihepatocellular carcinoma effects of (+)-cedrol, with mechanistic investigations demonstrating its ability to induce apoptosis, inhibit autophagy, and modulate the PI3K/AKT signalling pathway. This work underscores the utility of integrated biocatalytic design in enabling scalable production of bioactive phytochemicals while highlighting (+)-cedrol as a promising candidate for both mechanistic studies and therapeutic development