Molecular cloning and characterization of Pcal_0039, an ATP-/NAD
DNA ligase
Heterologous expression
Hyperthermophilic archaeon
Nick-joining
Pyrobaculum calidifontis
Journal
International journal of biological macromolecules
ISSN: 1879-0003
Titre abrégé: Int J Biol Macromol
Pays: Netherlands
ID NLM: 7909578
Informations de publication
Date de publication:
31 Dec 2023
31 Dec 2023
Historique:
received:
05
07
2023
revised:
25
08
2023
accepted:
03
09
2023
medline:
27
11
2023
pubmed:
7
9
2023
entrez:
6
9
2023
Statut:
ppublish
Résumé
The genome sequence of hyperthermophilic archaeon Pyrobaculum calidifontis contains an open reading frame, Pcal_0039, which encodes a putative DNA ligase. Structural analysis disclosed the presence of signature sequences of ATP-dependent DNA ligases. We have heterologously expressed Pcal_0039 gene in Escherichia coli. The recombinant protein, majorly produced in soluble form, was purified and functionally characterized. Recombinant Pcal_0039 displayed nick-joining activity between 40 and 85 °C. Optimal activity was observed at 70 °C and pH 5.5. Nick-joining activity was retained even after heating for 1 h at 90 °C, indicating highly thermostable nature of Pcal_0039. The nick-joining activity, displayed by Pcal_0039, was metal ion dependent and Mg
Identifiants
pubmed: 37673141
pii: S0141-8130(23)03608-5
doi: 10.1016/j.ijbiomac.2023.126711
pii:
doi:
Substances chimiques
NAD
0U46U6E8UK
DNA Ligase ATP
EC 6.5.1.1
DNA Ligases
EC 6.5.1.-
Adenosine Triphosphate
8L70Q75FXE
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
126711Informations de copyright
Copyright © 2023 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare that they have no competing interests.