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

Serra, Immacolata LU ; Ubiali, Daniela ; Piškur, Jure LU ; Munch-Petersen, Birgitte LU ; Bavaro, Teodora and Terreni, Marco (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
; ; ; ; and
organization
publishing date
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}},
}