Immobilized Drosophila melanogaster Deoxyribonucleoside Kinase (DmdNK) as a High Performing Biocatalyst for the Synthesis of Purine Arabinonucleotides
(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:
https://lup.lub.lu.se/record/4418878
- author
- Serra, Immacolata ; Conti, Silvia ; Piskur, Jure LU ; Clausen, Anders Ranegaard LU ; Munch-Petersen, Birgitte ; Terreni, Marco and Ubiali, Daniela
- organization
- publishing date
- 2014
- 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}}, }