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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)
Please use this url to cite or link to this publication:
author
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
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
2014-05-05 11:09:09
date last changed
2017-06-25 04:05:02
@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},
  keyword      = {biocatalysis,deoxyribonucleoside kinase,immobilization,nucleotides,phosphorylation},
  language     = {eng},
  number       = {2-3},
  pages        = {563--570},
  publisher    = {John Wiley & Sons},
  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},
  volume       = {356},
  year         = {2014},
}