2026-06-08T01:16:45Zhttps://docta.ucm.es/rest/oai/request

oai:docta.ucm.es:20.500.14352/1088672024-10-10T23:46:47Zcom_20.500.14352_14col_20.500.14352_15

Magnetic multi-enzymatic system for Cladribine manufacturing Cruz, Guillermo Saiz, Laura Pilar Bilal, Muhammad Eltoukhy, Lobna Loderer, Christoph Fernández Lucas, Jesús Enzyme-mediated processes have proven to be a valuable and sustainable alternative to traditional chemical methods. In this regard, the use of multi-enzymatic systems enables the realization of complex synthetic schemes, while also introducing a number of additional advantages, including the conversion of reversible reactions into irreversible processes, the partial or complete elimination of product inhibition problems, and the minimization of undesirable by-products. In addition, the immobilization of biocatalysts on magnetic supports allows for easy reusability and streamlines the downstream process. Herein we have developed a cascade system for cladribine synthesis based on the sequential action of two magnetic biocatalysts. For that purpose, purine 2′-deoxyribosyltransferase from Leishmania mexicana (LmPDT) and Escherichia coli hypoxanthine phosphoribosyltransferase (EcHPRT) were immobilized onto Ni2+-prechelated magnetic microspheres (MagReSyn®NTA). Among the resulting derivatives, MLmPDT3 (activity: 11,935 IU/gsupport, 63% retained activity, operational conditions: 40 °C and pH 5–7) and MEcHPRT3 (12,840 IU/gsupport, 45% retained activity, operational conditions: pH 5–8 and 40–60 °C) emerge as optimal catalysts for further synthetic application. Moreover, the MLmPDT3/MEcHPRT3 system was biochemically characterized and successfully applied to the one-pot synthesis of cladribine under various conditions. This methodology not only displayed a 1.67-fold improvement in cladribine synthesis (compared to MLmPDT3), but it also implied a practically complete transformation of the undesired by-product into a high-added-value product (90% conversion of Hyp into IMP). Finally, MLmPDT3/MEcHPRT3 was reused for 16 cycles, which displayed a 75% retained activity. 2024-10-10T15:27:13Z 2024-10-10T15:27:13Z 2024-10-10T15:27:13Z 2022-11 journal article https://hdl.handle.net/20.500.14352/108867 1422-0067 10.3390/ijms232113634 eng PID2020-117025RB-I00 info:eu-repo/grantAgreement/NCN//2020%2F37%2FK%2FST8%2F03805 info:eu-repo/grantAgreement/DFG//LO 2678%2F2-1 Cruz, Guillermo, et al. «Magnetic Multi-Enzymatic System for Cladribine Manufacturing». International Journal of Molecular Sciences, vol. 23, n.o 21, noviembre de 2022, p. 13634. DOI.org (Crossref), https://doi.org/10.3390/ijms232113634. http://creativecommons.org/licenses/by/4.0/ open access Attribution 4.0 International MDPI