期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 117, 期 -, 页码 17-24出版社
ELSEVIER
DOI: 10.1016/j.ijbiomac.2018.05.155
关键词
Endonuclease V; Thermococcus barophilus; DNA repair; Thermostability
资金
- National Natural Science Foundation of China [41306131]
- Provincial Natural Science Foundation Grant of Jiangsu Province, China [BK20130440]
- Academic Leader of Middle and Young People of Yangzhou University Grant
- practice innovation training program for college students in Jiangsu [201711117059Y]
Endonuclease V (Endo V) is an important enzyme for repairing deoxyinosine in DNA. While bacterial and eukaryotic endo Vs have been well studied, knowledge of archaeal endo Vs is limited. Here, we first presented biochemical characterization of a thermostable endonudease V from the hyperthermophilic euryarchaeon Thermococcus barophilus Ch5 (Tba endo V). The recombinant enzyme possessed optimal endonuclease activity for cleaving deoxyinosine-containing DNA at 70-90 degrees C. Furthermore, Tba endo V can withstand 100 degrees C for 120 min without significant loss of its activity, suggesting the enzyme is thermostable. Tba endo V exhibited varying cleavage efficiencies at various pH levels from 6.0 to 11.0, among which an optimal pH for the enzyme was 8.0-9.0. In addition, a divalent metal ion was required for the enzyme to cleave DNA. Mn2+ and Mg2+ were optimal ions for the enzyme's activity whereas Ca2+, Zn2+ and Co2+ inhibited the enzyme activity. Moreover, the enzyme activity was suppressed by high NaCI concentration. Tba endo V bound to all DNA substrates; however, the enzyme exhibited a higher affinity for binding to deoxyinosine-containing DNA than normal DNA. Our work provides valuable information for revealing the role of Tba endo V in the base excision repair pathway for deoxyinosine repair in Thermococcus. (C) 2018 Published by Elsevier B.V.
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