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Immobilized Drosophila melanogaster Deoxyribonucleoside Kinase (DmdNK) as a High Performing Biocatalyst for the Synthesis of Purine Arabinonucleotides

Serra, Immacolata ; Conti, Silvia ; Piskur, Jure LU ; Clausen, Anders Ranegaard LU ; Munch-Petersen, Birgitte ; Terreni, Marco and Ubiali, Daniela (2014) In Advanced Synthesis & Catalysis 356(2-3). p.563-570
Abstract
Fruit fly (Drosophila melanogaster) deoxyribonucleoside kinase (DmdNK; EC: 2.7.1.145) was characterized for its substrate specificity towards natural and non-natural nucleosides, confirming its potential in the enzymatic synthesis of modified nucleotides. DmdNK was adsorbed on a solid ion exchange support (bearing primary amino groups) achieving an expressed activity >98%. Upon cross-linking with aldehyde dextran, expressed activity was 30-40%. Both biocatalysts (adsorbed or cross-linked) were stable at pH10 and room temperature for 24h (about 70% of retained activity). The cross-linked DmdNK preparation was used for the preparative synthesis of arabinosyladenine monophosphate (araA-MP) and fludarabine monophosphate (FaraA-MP). Upon... (More)
Fruit fly (Drosophila melanogaster) deoxyribonucleoside kinase (DmdNK; EC: 2.7.1.145) was characterized for its substrate specificity towards natural and non-natural nucleosides, confirming its potential in the enzymatic synthesis of modified nucleotides. DmdNK was adsorbed on a solid ion exchange support (bearing primary amino groups) achieving an expressed activity >98%. Upon cross-linking with aldehyde dextran, expressed activity was 30-40%. Both biocatalysts (adsorbed or cross-linked) were stable at pH10 and room temperature for 24h (about 70% of retained activity). The cross-linked DmdNK preparation was used for the preparative synthesis of arabinosyladenine monophosphate (araA-MP) and fludarabine monophosphate (FaraA-MP). Upon optimization of the reaction conditions (50mM ammonium acetate, substrate/ATP ratio=1:1.25, 2mM MgCl2, 37 degrees C, pH8) immobilized DmdNK afforded the title nucleotides with high conversion (>90%), whereas with the soluble enzyme lower conversions were achieved (78-87%). Arabinosyladenine monophosphate was isolated in 95% yield and high purity (96.5%). (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
biocatalysis, deoxyribonucleoside kinase, immobilization, nucleotides, phosphorylation
in
Advanced Synthesis & Catalysis
volume
356
issue
2-3
pages
563 - 570
publisher
John Wiley & Sons Inc.
external identifiers
  • wos:000332000200038
  • scopus:84899910432
ISSN
1615-4150
DOI
10.1002/adsc.201300649
language
English
LU publication?
yes
id
20487de2-aa22-4d46-927c-2dcbad263a63 (old id 4418878)
date added to LUP
2016-04-01 14:09:18
date last changed
2022-01-27 23:01:23
@article{20487de2-aa22-4d46-927c-2dcbad263a63,
  abstract     = {{Fruit fly (Drosophila melanogaster) deoxyribonucleoside kinase (DmdNK; EC: 2.7.1.145) was characterized for its substrate specificity towards natural and non-natural nucleosides, confirming its potential in the enzymatic synthesis of modified nucleotides. DmdNK was adsorbed on a solid ion exchange support (bearing primary amino groups) achieving an expressed activity >98%. Upon cross-linking with aldehyde dextran, expressed activity was 30-40%. Both biocatalysts (adsorbed or cross-linked) were stable at pH10 and room temperature for 24h (about 70% of retained activity). The cross-linked DmdNK preparation was used for the preparative synthesis of arabinosyladenine monophosphate (araA-MP) and fludarabine monophosphate (FaraA-MP). Upon optimization of the reaction conditions (50mM ammonium acetate, substrate/ATP ratio=1:1.25, 2mM MgCl2, 37 degrees C, pH8) immobilized DmdNK afforded the title nucleotides with high conversion (>90%), whereas with the soluble enzyme lower conversions were achieved (78-87%). Arabinosyladenine monophosphate was isolated in 95% yield and high purity (96.5%).}},
  author       = {{Serra, Immacolata and Conti, Silvia and Piskur, Jure and Clausen, Anders Ranegaard and Munch-Petersen, Birgitte and Terreni, Marco and Ubiali, Daniela}},
  issn         = {{1615-4150}},
  keywords     = {{biocatalysis; deoxyribonucleoside kinase; immobilization; nucleotides; phosphorylation}},
  language     = {{eng}},
  number       = {{2-3}},
  pages        = {{563--570}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Advanced Synthesis & Catalysis}},
  title        = {{Immobilized Drosophila melanogaster Deoxyribonucleoside Kinase (DmdNK) as a High Performing Biocatalyst for the Synthesis of Purine Arabinonucleotides}},
  url          = {{http://dx.doi.org/10.1002/adsc.201300649}},
  doi          = {{10.1002/adsc.201300649}},
  volume       = {{356}},
  year         = {{2014}},
}