We here described a three-step multi-enzymatic reaction for the one-pot synthesis of vidarabine 5'-monophosphate (araA-MP), an antiviral drug, using arabinosyluracil (araU), adenine (Ade), and adenosine triphosphate (ATP) as precursors. To this aim, three enzymes involved in the biosynthesis of nucleosides and nucleotides were used in a cascade mode after immobilization: uridine phosphorylase from Clostridium perfringens (CpUP), a purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP), and deoxyadenosine kinase from Dictyostelium discoideum (DddAK). Specifically, CpUP catalyzes the phosphorolysis of araU thus generating uracil and α-d-arabinose-1-phosphate. AhPNP catalyzes the coupling between this latter compound and Ade to form araA (vidarabine). This nucleoside becomes the substrate of DddAK, which produces the 5'-mononucleotide counterpart (araA-MP) using ATP as the phosphate donor. Reaction conditions (i.e., medium, temperature, immobilization carriers) and biocatalyst stability have been balanced to achieve the highest conversion of vidarabine 5'-monophosphate (≥95.5%). The combination of the nucleoside phosphorylases twosome with deoxyadenosine kinase in a one-pot cascade allowed (i) a complete shift in the equilibrium-controlled synthesis of the nucleoside towards the product formation; and (ii) to overcome the solubility constraints of araA in aqueous medium, thus providing a new route to the highly productive synthesis of araA-MP.

Robescu, M., Serra, I., Terreni, M., Ubiali, D., Bavaro, T. (2020). A multi-enzymatic cascade reaction for the synthesis of vidarabine 5'-monophosphate. CATALYSTS, 10(1 (January 2020)) [10.3390/catal10010060].

A multi-enzymatic cascade reaction for the synthesis of vidarabine 5'-monophosphate

Serra I.
;
2020

Abstract

We here described a three-step multi-enzymatic reaction for the one-pot synthesis of vidarabine 5'-monophosphate (araA-MP), an antiviral drug, using arabinosyluracil (araU), adenine (Ade), and adenosine triphosphate (ATP) as precursors. To this aim, three enzymes involved in the biosynthesis of nucleosides and nucleotides were used in a cascade mode after immobilization: uridine phosphorylase from Clostridium perfringens (CpUP), a purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP), and deoxyadenosine kinase from Dictyostelium discoideum (DddAK). Specifically, CpUP catalyzes the phosphorolysis of araU thus generating uracil and α-d-arabinose-1-phosphate. AhPNP catalyzes the coupling between this latter compound and Ade to form araA (vidarabine). This nucleoside becomes the substrate of DddAK, which produces the 5'-mononucleotide counterpart (araA-MP) using ATP as the phosphate donor. Reaction conditions (i.e., medium, temperature, immobilization carriers) and biocatalyst stability have been balanced to achieve the highest conversion of vidarabine 5'-monophosphate (≥95.5%). The combination of the nucleoside phosphorylases twosome with deoxyadenosine kinase in a one-pot cascade allowed (i) a complete shift in the equilibrium-controlled synthesis of the nucleoside towards the product formation; and (ii) to overcome the solubility constraints of araA in aqueous medium, thus providing a new route to the highly productive synthesis of araA-MP.
Articolo in rivista - Articolo scientifico
Deoxyadenosine kinase; Enzyme cascade; Enzyme immobilization; Nucleoside phosphorylase; Phosphorylation; Transglycosylation; Unnatural nucleosides; Vidarabine 5′-monophosphate;
English
1-gen-2020
2020
10
1 (January 2020)
60
open
Robescu, M., Serra, I., Terreni, M., Ubiali, D., Bavaro, T. (2020). A multi-enzymatic cascade reaction for the synthesis of vidarabine 5'-monophosphate. CATALYSTS, 10(1 (January 2020)) [10.3390/catal10010060].
File in questo prodotto:
File Dimensione Formato  
catalysts 2020.pdf

accesso aperto

Tipologia di allegato: Publisher’s Version (Version of Record, VoR)
Dimensione 930.59 kB
Formato Adobe PDF
930.59 kB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/316392
Citazioni
  • Scopus 17
  • ???jsp.display-item.citation.isi??? 18
Social impact