Reprograming the\nCarbon Metabolism of Yeast for Hyperproducing\nMevalonate, a Building Precursor of the Terpenoid Backbone

Utilization of microbial hosts to produce natural plant\nproducts\nis regarded as a promising and sustainable approach. However, achieving\nhighly efficient production of terpenoids using microorganisms remains\na significant challenge. Here, mevalonate, a building block of terpenoids,\nwas used as a demo product to explore the potential metabolic constraints\nfor terpenoid biosynthesis in Yarrowia lipolytica. First, by regulation of the expression of <i>ERG12</i> and <i>HMGR</i>, the mevalonate titer was improved by\n7660%. Subsequently, the native mevalonate pathway (MVA pathway) was\nenhanced, and the production of mevalonate increased to 4.16 g/L.\nTo ensure a sufficient supply of acetyl-CoA, the citrate route and\nTCA cycle were simultaneously engineered, and the mevalonate titer\nwas further improved to 5.25 g/L in shake flasks. Ultimately, the\ncitrate overflow metabolism of Y. lipolytica was eliminated by deleting <i>CEX1</i>, resulting in the\nhighest mevalonate titer of 101 g/L with a yield of 0.255 g/g of glucose\nin eukaryotes. These insights could be applied to the effective production\nof terpenoids and biochemicals derived from central carbon metabolic\npathways.