Purine nucleoside phosphorylases (PNPs, EC 126.96.36.199) catalyze the reversible phosphorolysis of the glycosydic bond of purine nucleosides and, upon addition of a second nucleobase, may transfer the glycosyl moiety to stereoselectively form a new nucleoside (transglycosylation). Because of its wide substrate specificity,[1,2] a PNP from Aeromonas hydrophila (AhPNP) was exploited to catalyze a “one-pot, one-enzyme” batch transglycosylation, resulting in a moderate to high conversion (18-65%) of 6-substituted purine bases into a 23-compound library of 6-substituted purine-9-ribosides. Afterwards, AhPNP was covalently immobilized (50% yield) in a pre-packed stainless steel column and the resulting AhPNP-IMER (IMmobilized Enzyme Reactor) was coupled to a HPLC apparatus containing an analytical or semi-preparative chromatographic column associated with a UVvisible detector. The resulting system (Figure 1) was exploited to perform the flow-synthesis of five 6-substituted purine ribonucleosides at a mg scale by transglycosylation. Under optimized (Design of Experiments, DoE) reaction conditions, coupling of transglycosylation with product separation resulted in a fast and efficient process (52-89% conversion) with minimized sample handling. References:  Ubiali, D. et al. Adv. Synth. Catal. 2012, 354, 96-104.  Calleri, E. et al. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 2014, 968, 79-86.  Ubiali D. et al. Curr. Org. Chem. 2015, 19, 2220-2225.  Calleri, E. et al. Adv. Synth. Catal. 2015, 357, 2520-2528.
|Titolo:||Batch and flow synthesis of 6-substituted purine ribonucleosides by enzymatic transglycosylation|
|Data di pubblicazione:||13-giu-2016|
|Settore Scientifico Disciplinare:||Settore CHIM/06 - Chimica Organica|
|Citazione:||Batch and flow synthesis of 6-substituted purine ribonucleosides by enzymatic transglycosylation / M. Rabuffetti, G. Cattaneo, T. Bavaro, E. Calleri, G. Massolini, I. Serra, D. Ubiali, G. Speranza. ((Intervento presentato al 41. convegno International Summer School on Organic Synthesis "A. Corbella" ISOS 2016 tenutosi a Gargnano nel 2016.|
|Appare nelle tipologie:||14 - Intervento a convegno non pubblicato|