Flow‐Synthesis of Nucleosides Catalyzed by an Immobilized Purine Nucleoside Phosphorylase from Aeromonas hydrophila: Integrated Systems of Reaction Control and Product Purification. Issue 11 (10th August 2015)

• Record Type: Journal Article
• Authors: Calleri, Enrica; Cattaneo, Giulia; Rabuffetti, Marco; Serra, Immacolata; Bavaro, Teodora; Massolini, Gabriella; Speranza, Giovanna; Ubiali, Daniela
• Is Part Of: Advanced synthesis & catalysis. Volume 357:Issue 11(2015)
• Page Start: 2520
• Page End: 2528
• Publication Date: 2015-08-10
• Subjects: Catalysis -- Periodicals; Organic compounds -- Synthesis -- Periodicals; Chemistry -- Periodicals; Chemistry, Technical -- Periodicals; Chemistry -- Periodicals; Catalysis -- Periodicals; Technology, Pharmaceutical -- Periodicals; 547.2
• DOI: 10.1002/adsc.201500133
• Languages: English
• ISSNs: 1615-4150

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>A purine nucleoside phosphorylase from <italic>Aeromonas hydrophyla</italic> (<italic>Ah</italic>PNP) was covalently immobilized in a pre‐packed stainless steel column containing aminopropylsilica particles <italic>via</italic> Schiff base chemistry upon glutaraldehyde activation. The resulting <italic>Ah</italic>PNP‐IMER (<italic>Immobilized Enzyme Reactor</italic>, immobilization yield ≈50%) was coupled on‐line through a 6‐way switching valve to an HPLC apparatus containing an analytical or a semi‐preparative chromatographic column. The synthesis of five 6‐modified purine ribonucleosides was carried out by continuously pumping the reaction mixture through the <italic>Ah</italic>PNP‐IMER until the highest conversion was reached, and then directing the reaction mixture to chromatographic separation. The conditions of the <italic>Ah</italic>PNP‐catalyzed transglycosylations (2:1 <italic>ratio</italic> sugar <italic>donor</italic>:base <italic>acceptor</italic>; 10 mM phosphate buffer; pH 7.5; temperature 37 °C, flow rate 0.5 mL min<sup>−1</sup>) were optimized by a fractional factorial experimental design. Coupling the bioconversion step with the product purification in such an integrated platform resulted in a fast and efficient synthetic process (yield=52–89%; <10 mg) where sample handling was minimized. To date, <italic>Ah</italic>PNP‐IMER has retained completely its activity upon 50 reactions in 10 months.</p> <p> <graphic position="anchor" mimetype="image" xlink:href="ark:/27927/pgj2cqp61g8" orientation="portrait" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /> </p> </abstract>