RT Journal Article
T1 Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii
A1 Acosta, Javier
A1 Nguyen, Kim
A1 Spitale, Robert C.
A1 Fernández Lucas, Jesús
AB Nowadays, enzymatic synthesis of nucleotides is an efficient and sustainable alternative to chemical methodologies. In this regard, after the biochemical characterization of wild-type and mutant uracil phosphoribosyltransferases from Toxoplasma gondii (TgUPRT, TgUPRT2, and TgUPRT3), TgUPRT2 was selected as the optimal candidate (69.5 IU mg−1, UMP synthesis) for structure-guided immobilization onto Ni2+ chelate (MNiUPRT2) and onto glutaraldehyde-activated microparticles (MGlUPRT2). Among resulting derivatives, MNiUPRT23 (6127 IU g−1biocat; 92% retained activity; 3–5 fold enhanced stability at 50–60 °C) and MGlUPRT2N (3711 IU g−1biocat; 27% retained activity; 8–20 fold enhanced stability at 50–60 °C) displayed the best operability. Moreover, the enzymatic synthesis of different pyrimidine NMPs was performed. Finally, the reusability of both derivatives in 5-FUMP synthesis (MNiUPRT23, 80% retained activity after 7 cycles, 5 min; MGlUPRT2N, 70% retained activity after 10 cycles, 20 min) was carried out at short times.
PB Elsevier
SN 0960-8524
YR 2021
FD 2021-11
LK https://hdl.handle.net/20.500.14352/109276
UL https://hdl.handle.net/20.500.14352/109276
LA eng
NO Acosta J, Nguyen K, Spitale RC, Fernández-Lucas J. Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii. Bioresource Technology 2021;339:125649. https://doi.org/10.1016/j.biortech.2021.125649.
NO This work was also supported by grant 5R21MH116415 from National Institutes of Health (NIH)
NO Fundación Santander
NO Universidad Europea de Madrid
NO National Institutes of Health
DS Docta Complutense
RD 25 feb 2026