{Reference Type}: Journal Article
{Title}: Systematic Engineering of Escherichia coli for Efficient Production of Pseudouridine from Glucose and Uracil.
{Author}: Zhang C;Wei G;Zhou N;Wang Y;Feng J;Wang X;Zhang A;Chen K;
{Journal}: ACS Synth Biol
{Volume}: 13
{Issue}: 4
{Year}: 2024 04 19
{Factor}: 5.249
{DOI}: 10.1021/acssynbio.4c00028
{Abstract}: In this study, we proposed a biological approach to efficiently produce pseudouridine (Ψ) from glucose and uracil in vivo using engineered Escherichia coli. By screening host strains and core enzymes, E. coli MG1655 overexpressing Ψ monophosphate (ΨMP) glycosidase and ΨMP phosphatase was obtained, which displayed the highest Ψ concentration. Then, optimization of the RBS sequences, enhancement of ribose 5-phosphate supply in the cells, and overexpression of the membrane transport protein UraA were investigated. Finally, fed-batch fermentation of Ψ in a 5 L fermentor can reach 27.5 g/L with a yield of 89.2 mol % toward uracil and 25.6 mol % toward glucose within 48 h, both of which are the highest to date. In addition, the Ψ product with a high purity of 99.8% can be purified from the fermentation broth after crystallization. This work provides an efficient and environmentally friendly protocol for allowing for the possibility of Ψ bioproduction on an industrial scale.