Reusable and modular combinatorial libraries for iterative metabolic engineering of Saccharomyces cerevisiae

We developed a methodology for generating reusable and modular integration-based combinatorial libraries and demonstrate their usefulness by engineering red beet pigment production in the yeast Saccharomyces cerevisiae. Long-read amplicon sequencing of the integrated library DNA from the high-performing library members from each round revealed novel insights into yeast and betacyanin metabolism.