Metabolic engineering Saccharomyces cerevisiae for de novo biosynthesis of hydroxycinnamoyl glycerols

Hydroxycinnamoyl glycerols (HCGs) are a class of bioconjugates characterized by the esterification of hydroxycinnamic acids with glycerol. Owing to their diverse biological activities, HCGs are considered high-value materials with potential applications in the food, pharmaceutical, and cosmetics industries. In this study, a de novo biosynthetic pathway of HCGs was constructed by integrating a p-coumaric acid-producing module and a 4-coumarate:coenzyme A ligase - hydroxycinnamoyltransferase (4CL-HCT) module into the genome of Saccharomyces cerevisiae. Through optimization of the primary metabolic pathway, fine-tuning of the shikimate pathway, and strategic selection of integration sites, the upstream metabolic flux in the S. cerevisiae strain was engineering to enhance the availability of l-tyrosine and l-phenylalanine precursors, as well as acetyl-coenzyme A (acetyl-CoA) and adenosine triphosphate (ATP) supply. As a result, the final engineered S. cerevisiae strain produced 1-O-p-coumaroylglycerol (1-OPCG) at a titer of 8.49 ± 2.29 μg/L in shake-flask fermentation. The strategies employed in this study provide a robust foundation for the synthesis of other HCGs.