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

Resumé

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

Citer dette

@article{17f4da056e6347b2b32f591240d91df6,
title = "Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine",
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.",
author = "Immacolata Serra and Daniela Ubiali and Jure Piskur and Birgitte Munch-Petersen and Teodora Bavaro and Marco Terreni",
year = "2017",
doi = "10.1002/slct.201700558",
language = "English",
volume = "2",
pages = "5403--5408",
journal = "Chemistry Select",
issn = "2365-6549",
publisher = "Wiley-VCH",
number = "19",

}

Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine. / Serra, Immacolata ; Ubiali, Daniela; Piskur, Jure; Munch-Petersen, Birgitte; Bavaro, Teodora; Terreni, Marco.

I: Chemistry Select, Bind 2, Nr. 19, 2017, s. 5403-5408.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Immobilization of Deoxyadenosine Kinase from Dictyostelium discoideum (DddAK) and Its Application in the 5’-Phosphorylation of Arabinosyladenine and Arabinosyl-2-fluoroadenine

AU - Serra, Immacolata

AU - Ubiali, Daniela

AU - Piskur, Jure

AU - Munch-Petersen, Birgitte

AU - Bavaro, Teodora

AU - Terreni, Marco

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

U2 - 10.1002/slct.201700558

DO - 10.1002/slct.201700558

M3 - Journal article

VL - 2

SP - 5403

EP - 5408

JO - Chemistry Select

JF - Chemistry Select

SN - 2365-6549

IS - 19

ER -