In nature, enzyme cascades can be found in many metabolic pathways. The idea of using multienzymatic systems to mimic these processes is gaining interest for production of chemical compounds. A type of multi-enzymatic application is the use of multiple enzymes for shifting reaction equilibria. This strategy relies on removing intermediates, inhibitory products or byproducts, via a second enzymatic reaction. In the context of a multi-enzymatic system, a one-pot process uses more than one enzyme in a single reactor.1 We here describe a three-step sequential 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 immobilized biocatalysts involved in the biosynthesis of nucleosides and nucleotides were used: uridine phosphorylase from Clostridium perfringens (CpUP),2 a purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP),2 and deoxyadenosine kinase from Dictyostelium discoideum (DddAK).3 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 (Scheme 1). Reaction conditions (i.e. medium, temperature, immobilization carriers) and biocatalyst stability have been balanced and optimized to achieve the highest productivity. Vidarabine 5’-monophosphate was obtained in 95.5% conversion. Optimization of the purification step is in progress.

A green synthesis of vidarabine 5’-monophosphate via a one-pot multienzymatic reaction catalyzed by immobilized biocatalysts

UBIALI, DANIELA;CATTANEO, GIULIA;BAVARO, TEODORA;TERRENI, MARCO;SERRA, IMMACOLATA
2015-01-01

Abstract

In nature, enzyme cascades can be found in many metabolic pathways. The idea of using multienzymatic systems to mimic these processes is gaining interest for production of chemical compounds. A type of multi-enzymatic application is the use of multiple enzymes for shifting reaction equilibria. This strategy relies on removing intermediates, inhibitory products or byproducts, via a second enzymatic reaction. In the context of a multi-enzymatic system, a one-pot process uses more than one enzyme in a single reactor.1 We here describe a three-step sequential 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 immobilized biocatalysts involved in the biosynthesis of nucleosides and nucleotides were used: uridine phosphorylase from Clostridium perfringens (CpUP),2 a purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP),2 and deoxyadenosine kinase from Dictyostelium discoideum (DddAK).3 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 (Scheme 1). Reaction conditions (i.e. medium, temperature, immobilization carriers) and biocatalyst stability have been balanced and optimized to achieve the highest productivity. Vidarabine 5’-monophosphate was obtained in 95.5% conversion. Optimization of the purification step is in progress.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11571/1180306
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