Biochemical Characterization of an Arabinoside Monophosphate Specific 5'-Nucleotidase-like Enzyme from

Overview

Journal Biomolecules

Publisher MDPI

Specialties Biochemistry
Molecular Biology

Date 2024 Nov 27

PMID 39595545

Authors

Yuxue Liu

Xiaobei Liu

Xiaojing Zhang

Xiaoting Tang

Weiwei Su

Zhenyu Wang

Hailei Wang

Affiliations

Soon will be listed here.

Abstract

To investigate the function of the gene in the pentostatin and vidarabine (Ara-A) biosynthetic gene cluster in NRRL 3238, PenF was recombinantly expressed and characterized. Enzymatic characterization of the enzyme demonstrated that PenF exhibited metal-dependent nucleoside 5'-monophosphatase activity, showing a substrate preference for arabinose nucleoside 5'-monophosphate over 2'-deoxyribonucleoside 5'-monophosphate and ribonucleoside 5'-monophosphate. Metal ions such as Mg and Mn significantly enhanced enzyme activity, whereas Zn, Cu, and Ca inhibited it. For vidarabine 5'-monophosphate, the and values were determined to be 71.5 μM and 33.9 min, respectively. The value was 474.1 mM·min for vidarabine 5-monophosphate and was 68-fold higher than that for 2'-deoxyadenosine 5'-monophosphate. Comparative sequence alignment and structural studies suggested that residues outside the primary substrate-binding site are responsible for this substrate specificity. In conclusion, PenF's activity toward vidarabine 5'-monophosphate likely plays a role in the dephosphorylation of precursors during Ara-A biosynthesis.

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