Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine
(2017) In ChemistrySelect 2(19). p.5403-5408- 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... (More)
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.
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- author
- Serra, Immacolata LU ; Ubiali, Daniela ; Piškur, Jure LU ; Munch-Petersen, Birgitte LU ; Bavaro, Teodora and Terreni, Marco
- organization
- publishing date
- 2017
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- arabinonucleoside-5’-monophosphate, deoxyadenosine kinase, immobilization, nucleotide synthesis, phosphorylation
- in
- ChemistrySelect
- volume
- 2
- issue
- 19
- pages
- 6 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:85041745989
- ISSN
- 2365-6549
- DOI
- 10.1002/slct.201700558
- language
- English
- LU publication?
- yes
- id
- df4741b6-e58a-45e2-a27c-612f619e5232
- date added to LUP
- 2018-02-22 13:23:30
- date last changed
- 2022-04-25 05:49:28
@article{df4741b6-e58a-45e2-a27c-612f619e5232, abstract = {{<p>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 (Sepabeads<sup>TM</sup> 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 POCl<sub>3</sub>-based phosphorylation.</p>}}, author = {{Serra, Immacolata and Ubiali, Daniela and Piškur, Jure and Munch-Petersen, Birgitte and Bavaro, Teodora and Terreni, Marco}}, issn = {{2365-6549}}, keywords = {{arabinonucleoside-5’-monophosphate; deoxyadenosine kinase; immobilization; nucleotide synthesis; phosphorylation}}, language = {{eng}}, number = {{19}}, pages = {{5403--5408}}, publisher = {{Wiley-Blackwell}}, series = {{ChemistrySelect}}, title = {{Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine}}, url = {{http://dx.doi.org/10.1002/slct.201700558}}, doi = {{10.1002/slct.201700558}}, volume = {{2}}, year = {{2017}}, }