A modular in vitro multienzyme cascade for the sustainable synthesis of high-value vidarabine 5′-monophosphate from adenosine
- 1Henan Engineering Center of Bioconversion Technology of Functional Microbes, College of Life Science, Henan Normal University, Xinxiang, 453007, People's Republic of China
- 2MOE Key Laboratory of Bio-Intelligent Manufacturing, School of Bioengineering, Dalian University of Technology, Dalian, 116024, People's Republic of China
Abstract
Nucleoside analogues such as vidarabine (Ara-A) are vital therapeutics but are often limited by poor pharmacokinetics. Its phosphorylated form, vidarabine 5′-monophosphate (Ara-AMP), offers improved solubility and is used as a clinical alternative, although it can be dephosphorylated to Ara-A in vivo. Here, we report a modular multienzyme cascade for the sustainable synthesis of Ara-AMP directly from adenosine. The cascade integrates three functional modules: a core module (PNP, PPM, and RpiA) for the conversion of adenosine to Ara-A, a driver module (dNKδ20) for the selective phosphorylation of Ara-A to Ara-AMP, and a support module (PPK) for ATP regeneration from low-cost ADP and polyphosphate. The use of dNKδ20, a mutant deoxyribonucleoside kinase with high selectivity for Ara-A over adenosine, is critical for minimizing the formation of AMP byproduct, which would otherwise complicate downstream processing. Under optimized conditions, the cascade achieved an 80.2% conversion of 1 mM adenosine to Ara-AMP in 4 h using 0.05 mM ADP. The modular design was also applicable to guanosine and inosine, affording the corresponding arabinosyl nucleotides, albeit with lower yields. This work establishes a sustainable biocatalytic platform for nucleoside analogues, exemplifying how enzyme promiscuity, thermodynamic driving, and cofactor recycling enable green pharmaceutical manufacturing.
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BibTeX
@article{su2026modular,
title = {{A modular in vitro multienzyme cascade for the sustainable synthesis of high-value vidarabine 5′-monophosphate from adenosine}},
author = {Weiwei Su and Xiaofan Zhang and Rongrong Li and Kaige Li and Xiaojia Guo and Xiaobei Liu and Zhenyu Wang and Hailei Wang and Yuxue Liu},
journal = {Synthetic and Systems Biotechnology},
year = {2026},
month = oct,
doi = {10.1016/j.synbio.2026.10.005},
url = {https://doi.org/10.1016/j.synbio.2026.10.005},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - A modular in vitro multienzyme cascade for the sustainable synthesis of high-value vidarabine 5′-monophosphate from adenosine
AU - Weiwei Su
AU - Xiaofan Zhang
AU - Rongrong Li
AU - Kaige Li
AU - Xiaojia Guo
AU - Xiaobei Liu
AU - Zhenyu Wang
AU - Hailei Wang
AU - Yuxue Liu
PY - 2026
DA - 2026/10/16
JO - Synthetic and Systems Biotechnology
DO - 10.1016/j.synbio.2026.10.005
UR - https://doi.org/10.1016/j.synbio.2026.10.005
AB - Nucleoside analogues such as vidarabine (Ara-A) are vital therapeutics but are often limited by poor pharmacokinetics. Its phosphorylated form, vidarabine 5′-monophosphate (Ara-AMP), offers improved solubility and is used as a clinical alternative, although it can be dephosphorylated to Ara-A in vivo. Here, we report a modular multienzyme cascade for the sustainable synthesis of Ara-AMP directly from adenosine. The cascade integrates three functional modules: a core module (PNP, PPM, and RpiA) for the conversion of adenosine to Ara-A, a driver module (dNKδ20) for the selective phosphorylation of Ara-A to Ara-AMP, and a support module (PPK) for ATP regeneration from low-cost ADP and polyphosphate. The use of dNKδ20, a mutant deoxyribonucleoside kinase with high selectivity for Ara-A over adenosine, is critical for minimizing the formation of AMP byproduct, which would otherwise complicate downstream processing. Under optimized conditions, the cascade achieved an 80.2% conversion of 1 mM adenosine to Ara-AMP in 4 h using 0.05 mM ADP. The modular design was also applicable to guanosine and inosine, affording the corresponding arabinosyl nucleotides, albeit with lower yields. This work establishes a sustainable biocatalytic platform for nucleoside analogues, exemplifying how enzyme promiscuity, thermodynamic driving, and cofactor recycling enable green pharmaceutical manufacturing.
SN - 2405-805X
ER -