Multiomics Elucidation and Engineering Optimization of Efficient Pterostilbene Biosynthesis in <i>Escherichia coli</i>

Pterostilbene is a bioactive compound with diverse health-promoting properties and has garnered growing interest in biomedical and industrial applications. However, traditional production methods based on plant extraction or chemical synthesis are limited by a low yield, high cost, and sustainability concerns. Microbial fermentation therefore represents a promising alternative for scalable and ecofriendly pterostilbene production. In this study, a comprehensive multiomics analysis was conducted to elucidate the mechanisms underlying efficient de novo pterostilbene biosynthesis in engineered Escherichia coli. The high-producing strain exhibited enhanced energy generation, improved precursor and cofactor availability, and a reprogrammed oxidative stress response. Guided by these omics insights, targeted genetic modifications further increased the pterostilbene yield, with the optimized strain achieving a titer of 150.47 mg/L, which is the highest de novo production reported to date in a bacterial system. These findings highlight the potential of E. coli as a robust platform for the biosynthesis of value-added natural products and provide a rational framework for engineering microbial cell factories optimized for biosynthetic pathways that are both energetically demanding and redox-regulated.