RT Journal Article
T1 Enzymatic production of non-natural nucleoside-5′-monophosphates by a thermostable uracil phosphoribosyltransferase
A1 Del Arco, Jon
A1 Acosta, Javier
A1 Pereira, Humberto M.
A1 Perona Requena, Almudena
A1 Lokanath, Neratur K.
A1 Kunishima, Naoki
A1 Fernández Lucas, Jesús
AB The use of enzymes as biocatalysts applied to synthesis ofmodified nucleoside-5’-monophosphates (NMPs) is an interest-ing alternative to traditional multistep chemical methodswhich offers several advantages, such as stereo-, regio-, andenantioselectivity, simple downstream processing, and mild re-action conditions. Herein we report the recombinant expres-sion, production, and purification of uracil phosphoribosyl-transferase from Thermus themophilus HB8 (TtUPRT). The struc-ture of TtUPRT has been determined by protein crystallogra-phy, and its substrate specificity and biochemical characteris-tics have been analyzed, providing new structural insights intothe substrate-binding mode. Biochemical characterization ofthe recombinant protein indicates that the enzyme is a homo-tetramer, with activity and stability across a broad range oftemperatures (50–80 8C), pH (5.5–9) and ionic strength (0–500 mm NaCl). Surprisingly, TtUPRT is able to recognize several5 and 6-substituted pyrimidines as substrates. These experi-mental results suggest TtUPRT could be a valuable biocatalystfor the synthesis of modified NMPs.
PB Wiley
SN 1867-3880
YR 2018
FD 2018
LK https://hdl.handle.net/20.500.14352/110029
UL https://hdl.handle.net/20.500.14352/110029
LA eng
NO del Arco, J., Acosta, J., Pereira, H. M., Perona, A., Lokanath, N. K., Kunishima, N., & Fernández-Lucas, J. (2018). Enzymatic Production of Non-Natural Nucleoside-5′-Monophosphates by a Thermostable Uracil Phosphoribosyltransferase. ChemCatChem, 10(2), 439-448. https://doi.org/10.1002/CCTC.201701223
NO Fundación Santander
NO Universidad Europea de Madrid
DS Docta Complutense
RD 19 abr 2025