Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine

Immacolata Serra, Daniela Ubiali, Jure Piskur, Birgitte Munch-Petersen, Teodora Bavaro, Marco Terreni

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Abstract

Deoxyadenosine kinase from Dictyostelium discoideum (DddAK) phosphorylates its natural substrate (2’‐deoxyadenosine, dAdo) as well as the arabinosyladenine analogues vidarabine (araA) and fludarabine (F‐araA) to their corresponding 5’‐monophosphates. DddAK has been here immobilized by ionic interaction on an aminated epoxy‐functionalized support (SepabeadsTM EC‐EP), and cross‐linked with oxidized dextran. The final activity recovery was 33–42 %, depending on the protein loading. Immobilization enhanced the stability of DddAK at pH 10 and, to a lesser extent, at 45 °C. Phosphorylation of dAdo, araA and F‐araA catalyzed by immobilized DddAK was always nearly quantitative in less than 12 hours. Fludarabine monophosphate was synthesized from F‐araA (95 % conversion, 20 g/L) using immobilized DddAK in fully aqueous medium, thus showing that this biocatalyst could be used for developing a preparative phosphorylation process greener and more efficient than POCl3‐based phosphorylation.
OriginalsprogEngelsk
TidsskriftChemistry Select
Vol/bind2
Udgave nummer19
Sider (fra-til)5403-5408
ISSN2365-6549
DOI
StatusUdgivet - 2017

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