Enhanced production of 3-hydroxypropionic acid from glucose and xylose by alleviation of metabolic congestion due to glycerol flux in engineered Escherichia coli

•A metabolomic study revealed the metabolic burden during glucose and xylose co-fermentation.•Replacement of the GPD1 gene with the gpsA gene alleviated metabolic congestion.•The carbon flux to 3-HP production was improved in the newly engineered strain.•Fed-batch fermentation of the glucose/xylose mixture produced 37.6 g/L 3-HP titer. Among platform chemicals obtained from renewable biomass, 3-hydroxypropionic acid (3-HP) has attracted considerable attention. A GC/TOF-MS study revealed that the intracellular metabolites of the TCA cycle and fatty acid synthesis increased in JHS01302, a galP-overexpressing strain of Escherichia coli, during glucose and xylose co-fermentation. Decreased intracellular glycerol levels and increased intracellular biosynthesis of 3-HP were also detected in the strain. Based on these results, the yeast GPD1 gene was replaced with the endogenous gpsA gene to modulate the rate of glycerol metabolism. In flask cultures, JHS01304 containing the gpsA gene displayed 43% lower glycerol accumulation and 52% higher 3-HP production than the control. JHS01304 produced 37.6 g/L 3-HP with a productivity rate of 0.63 g/L/h and yield of 0.17 g/g in the fed-batch fermentation. The metabolome analysis provided valuable information for alleviating the metabolic burden of glycerol flux to improve the production of 3-HP during glucose and xylose co-fermentation.