Semi-enzymatic synthesis of pseudouridine

Metabolic Engineering of Escherichia coli for Efficient Production of Pseudouridine

“…In the initial 8 h, the cells grew exponentially, but no product accumulated. Isopropyl-β- d -thiogalactopyranoside (IPTG) was added to the medium after 8 h of cultivation, and the fermentation temperature was decreased to 30 °C, which was optimal for ΨMP glycosidase synthesis . Then, pseudouridine began to accumulate.…”

“…The ΨMP synthesis reaction involving ΨMP glycosidase was favored for pseudouridine production and was promoted by ΨMP dephosphorylation. , Therefore, the ΨMP glycosidase and phosphatase genes were the focus of this study. First, the phosphatase genes were screened to determine the most suitable genes for pseudouridine production.…”

“…psuG has bidirectional catalysis activity; it can hydrolyze ΨMP to uracil and ribose 5-phosphate, and it can also catalyze the reverse reaction to synthesize pseudouridine and other xenobiotic nucleic acids. , It is widely present in the genomes of prokaryotes . In this study, EcpsuG and four other psuG genes from different sources were tested: KspsuG (from Klebsiella spallanzanii), RspsuG (from Rhizobium sp.…”

“…Uracil and ribose 5′-phosphate were catalyzed by pseudouridine 5′-phosphate glycosidase (psuG) to synthesize pseudouridine 5′-phosphate, and pseudouridine-5′-phosphate was dephosphorylated by alkaline phosphatase (phoA) to obtain pseudouridine (Figure A) . A semi-enzymatic synthesis approach was developed for the synthesis of pseudouridine . In this approach, the substrate ribose 5′-phosphate was hydrolyzed from adenosine 5′-monophosphate.…”

C-Ribosylating Enzymes in the (Bio)Synthesis of C-Nucleosides and C-Glycosylated Natural Products

“…ΨMPGs have drawn interest for use in biocatalytic synthesis. ,,,,, The enzymes appear to be convenient to work with in respect to expression in standard hosts (e.g., E. coli BL21), specific activity, and general stability. ,,,, Their substrates [e.g., Rib5P ( 22 ), uracil ( 23 ), and polyketides] are usually available as reagents for small-scale reactions. Aside from divalent metals (Mg 2+ , Mn 2+ ) that are added at low mM concentration, the enzymatic reactions do not require complex supplementation. ,,, This facilitates product recovery. Mesophilic enzymes enable reactions under conditions (≤40 °C, neutral pH) that prevent uncatalyzed side reactions on substrates [e.g., decomposition of Rib5P ( 22 )] and products …”