Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-9521
Main Title: Human Deoxycytidine Kinase Is a Valuable Biocatalyst for the Synthesis of Nucleotide Analogues
Author(s): Hellendahl, Katja F.
Kamel, Sarah
Wetterwald, Albane
Neubauer, Peter
Wagner, Anke
Type: Article
Language Code: en
Abstract: Natural ribonucleoside-5’-monophosphates are building blocks for nucleic acids which are used for a number of purposes, including food additives. Their analogues, additionally, are used in pharmaceutical applications. Fludarabine-5´-monophosphate, for example, is effective in treating hematological malignancies. To date, ribonucleoside-5’-monophosphates are mainly produced by chemical synthesis, but the inherent drawbacks of this approach have led to the development of enzymatic synthesis routes. In this study, we evaluated the potential of human deoxycytidine kinase (HsdCK) as suitable biocatalyst for the synthesis of natural and modified ribonucleoside-5’-monophosphates from their corresponding nucleosides. Human dCK was heterologously expressed in E. coli and immobilized onto Nickel-nitrilotriacetic acid (Ni-NTA) superflow. A screening of the substrate spectrum of soluble and immobilized biocatalyst revealed that HsdCK accepts a wide range of natural and modified nucleosides, except for thymidine and uridine derivatives. Upon optimization of the reaction conditions, HsdCK was used for the synthesis of fludarabine-5´-monophosphate using increasing substrate concentrations. While the soluble biocatalyst revealed highest product formation with the lowest substrate concentration of 0.3 mM, the product yield increased with increasing substrate concentrations in the presence of the immobilized HsdCK. Hence, the application of immobilized HsdCK is advantageous upon using high substrate concentration which is relevant in industrial applications.
URI: https://depositonce.tu-berlin.de/handle/11303/10595
http://dx.doi.org/10.14279/depositonce-9521
Issue Date: 27-Nov-2019
Date Available: 13-Jan-2020
DDC Class: 660 Chemische Verfahrenstechnik
Subject(s): human deoxycytidine kinase
Ni-NTA sepharose
activity screening
nucleoside analogue
cladribine
clofarabine
fludarabine
Sponsor/Funder: DFG, 392246628, Chemo-enzymatische Synthese von Selen-modifizierten Nukleosiden, Nukleotiden und Oligonukleotiden
License: https://creativecommons.org/licenses/by/4.0/
Journal Title: Catalysts
Publisher: MDPI
Publisher Place: Basel
Volume: 9
Issue: 12
Article Number: 997
Publisher DOI: 10.3390/catal9120997
EISSN: 2073-4344
Appears in Collections:FG Bioverfahrenstechnik » Publications

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