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

Immobilized Drosophila melanogaster Deoxyribonucleoside Kinase (DmdNK) as a High Performing... 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 pH 10 and room temperature for 24 h (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 (50 mM ammonium acetate, substrate/ATP ratio=1:1.25, 2 mM MgCl2, 37 °C, pH 8) 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%). http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Advanced Synthesis & Catalysis Wiley

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

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Publisher
Wiley
Copyright
Copyright © 2014 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1615-4150
eISSN
1615-4169
DOI
10.1002/adsc.201300649
Publisher site
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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 pH 10 and room temperature for 24 h (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 (50 mM ammonium acetate, substrate/ATP ratio=1:1.25, 2 mM MgCl2, 37 °C, pH 8) 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%).

Journal

Advanced Synthesis & Catalysis – Wiley

Published: Oct 10, 2014

Keywords: ; ; ; ;

References