Orotic acid production from crude glycerol by engineered Ashbya gossypii

Orotic acid is an intermediate of the de novo pyrimidine biosynthesis that plays a crucial role in several nutraceutical supplements. In some microorganisms, blockage this pathway downstream dihydroorotate dehydrogenase (DHOD) leads to orotic accumulation. Here, Ashbya gossypii Agura3 auxotroph was shown accumulate and excrete (~1.5 g/L), trait further explored for production from raw substrates. Metabolic engineering strain combined with culture conditions optimization led 3.6-fold increase crude glycerol. The mitochondrial DHOD encoded by AgURA9 be determinant phenotype. Heterologous expression engineered S. cerevisiae enabled accumulation excretion (~0.5 g/L). This study demonstrates potential A. valorization glycerol discloses molecular determinants its fungi.