Sun, Y.; Chen, C.; Lin, C.; Zhang, H.; Lian, J.; Hong, B.

Glyceollins are phytoalexins produced by soybeans in response to stressors such as pathogen invasion, injury, and envi-ronmental challenges. These compounds have attracted significant attention for their potential anticancer, anti-inflammatory, and antioxidant properties. However, their production is limited by the high cost of chemical synthesis, purification from soybeans, and incomplete knowledge of their biosynthetic pathways hinder their production via syn-thetic biology. In this study, we identified previously uncharacterized genes encoding reductase for 7,2\',4\'-trihydroxyisoflavanol (THI) synthesis and P450 enzymes responsible for the final oxidative cycliaztions in glyceollins I, II, and III biosynthesis, thereby completing the entire biosynthetic pathway. By reconstructing the pathway de novo through synthetic biology, we achieved the successful production of glyceollins from simple carbon sources in Baker\'s yeast. This work advances understanding of glyceollin biosynthesis, facilitates sustainable production in microbial hosts and offers new opportunities for their applications in agriculture and biology.