RT Journal Article
T1 Structural and functional characterization of thermostable biocatalysts for the synthesis of 6-aminopurine nucleoside-5′-monophospate analogues
A1 Del Arco, Jon
A1 Pérez, Elena
A1 Naitow, Hisashi
A1 Matsuura, Yoshinori
A1 Kunishima, Naoki
A1 Fernández Lucas, Jesús
AB The present work describes the functional and structural characterization of adenine phosphoribosyltransferase 2 from Thermus thermophilus HB8 (TtAPRT2). The combination of structural and substrate specificity data provided valuable information for immobilization studies. Dimeric TtAPRT2 was immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles by two different strategies: a) an enzyme immobilization at pH 8.5 to encourage the immobilization process by N-termini (MTtAPRT2A, MTtAPRT2B, MTtAPRT2C) or b) an enzyme immobilization at pH 10.0 to encourage the immobilization process through surface exposed lysine residues (MTtAPRT2D, MTtAPRT2E, MTtAPRT2F). According to catalyst load experiments, MTtAPRT2B (activity: 480 IU g−1biocatalyst, activity recovery: 52%) and MTtAPRT2F (activity: 507 IU g−1biocatalyst, activity recovery: 44%) were chosen as optimal derivatives. The biochemical characterization studies demonstrated that immobilization process improved the thermostability of TtAPRT2. Moreover, the potential reusability of MTtAPRT2B and MTtAPRT2F was also tested. Finally, MTtAPRT2F was employed in the synthesis of nucleoside-5′-monophosphate analogues.
PB Elsevier
SN 0960-8524
YR 2019
FD 2019-03
LK https://hdl.handle.net/20.500.14352/109898
UL https://hdl.handle.net/20.500.14352/109898
LA eng
NO Arco, JD, Pérez, E., Naitow, H., Matsuura, Y., Kunishima, N. y Fernández-Lucas, J. (2019). Caracterización estructural y funcional de biocatalizadores termoestables para la síntesis de análogos de nucleósido-5''-monofosfato de 6-aminopurina. Tecnología de recursos biológicos , 276 , 244–252. https://doi.org/10.1016/J.BIORTECH
NO Fundación Santander
NO Universidad Europea de Madrid
DS Docta Complutense
RD 7 abr 2025