The study describes the de novo biosynthesis of xanthohumol, a bioactive flavonoid found in hops, in *Saccharomyces cerevisiae*. Xanthohumol is a valuable compound with various health benefits but is unstable and present in low concentrations in beer. The researchers engineered *S. cerevisiae* to produce xanthohumol by balancing three parallel biosynthetic pathways: p-coumaric-CoA (p-CA-CoA) biosynthesis, malonyl-CoA supply, and the mevalonate (MVA) pathway. They enhanced the prenylation step, which is crucial for xanthohumol production, by engineering prenyltransferase (PTase) and improving the availability of dimethylallyl diphosphate (DMAPP). This approach increased the production of the key precursor demethylxanthohumol (DMX) by 83-fold. Additionally, they optimized the methylation step to achieve the de novo biosynthesis of xanthohumol. The study provides a systematic approach for engineering yeast cell factories to produce complex natural products, highlighting the potential for sustainable and economically viable production of valuable compounds.The study describes the de novo biosynthesis of xanthohumol, a bioactive flavonoid found in hops, in *Saccharomyces cerevisiae*. Xanthohumol is a valuable compound with various health benefits but is unstable and present in low concentrations in beer. The researchers engineered *S. cerevisiae* to produce xanthohumol by balancing three parallel biosynthetic pathways: p-coumaric-CoA (p-CA-CoA) biosynthesis, malonyl-CoA supply, and the mevalonate (MVA) pathway. They enhanced the prenylation step, which is crucial for xanthohumol production, by engineering prenyltransferase (PTase) and improving the availability of dimethylallyl diphosphate (DMAPP). This approach increased the production of the key precursor demethylxanthohumol (DMX) by 83-fold. Additionally, they optimized the methylation step to achieve the de novo biosynthesis of xanthohumol. The study provides a systematic approach for engineering yeast cell factories to produce complex natural products, highlighting the potential for sustainable and economically viable production of valuable compounds.