The complete synthetic pathway of echinacoside from Cistanche deserticola and its de novo biosynthesis in yeast

Echinacoside (ECH), a representative phenylethanol glycoside, possesses diverse pharmacological properties and has been employed in the treatment of neurodegenerative disorders (e.g., Parkinson's and Alzheimer's diseases), ischemic brain injury, and cancer. The growing therapeutic demand for ECH has highlighted the need for scalable production. However, conventional methods face significant challenges: chemical synthesis is hindered by the compound's structural complexity, while plant extraction is limited by the low natural yield of ECH from Cistanche deserticola, a parasitic desert plant with host-dependent growth. To establish a sustainable microbial biosynthesis platform, we first deciphered the biosynthetic pathway of ECH in C. deserticola by integrating metabolomic analyses of plant tissues and callus cultures. This enabled the identification of key precursors, enzymatic steps, and regulatory mechanisms. Leveraging this knowledge, we engineered Saccharomyces cerevisiae for de novo ECH production by reconstructing the heterologous pathway, achieving a titer of 7.52 ± 1.42 mg/L. Our study not only provides a foundation for the industrial-scale production of ECH but also deepens the understanding of bioactive compound biosynthesis in parasitic plants, offering insights for future pathway engineering efforts.