|Biochemical characterization of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2|
|Zhang, LK; Huang, YC; Xu DD(徐丹丹); Gong Y(龚勇); Xu, DD; Yang, LX; Qian, KC; Chang, GZ; Gong, Y; Zhou, XJ; Ma, KS
|发表期刊||APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
|摘要||His-Asn-His (HNH) proteins are a very common family of small nucleic acid-binding proteins that are generally associated with endonuclease activity and are found in all kingdoms of life. Although HNH endonucleases from mesophiles have been widely investigated, the biochemical functions of HNH endonucleases from thermophilic bacteriophages remain unknown. Here, we characterized the biochemical properties of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2. The recombinant GVE2 HNH endonuclease exhibited non-specific cleavage activity at high temperature. The optimal temperature of the GVE2 HNH endonuclease for cleaving DNA was 60-65 A degrees C, and the enzyme retained its DNA cleavage activity even after heating at 100 A degrees C for 30 min, suggesting the enzyme is a thermostable endonuclease. The GVE2 HNH endonuclease cleaved DNA over a wide pH spectrum, ranging from 5.5 to 9.0, and the optimal pH for the enzyme activity was 8.0-9.0. Furthermore, the GVE2 HNH endonuclease activity was dependent on a divalent metal ion. While the enzyme is inactive in the presence of Cu2+, the GVE2 HNH endonuclease displayed cleavage activity of varied efficiency with Mn2+, Mg2+, Ca2+, Fe2+, Co2+, Zn2+, and Ni2+. The GVE2 HNH endonuclease activity was inhibited by NaCl. This study provides the basis for determining the role of this endonuclease in life cycle of the bacteriophage GVE2 and suggests the potential application of the enzyme in molecular biology and biotechnology.|
Divalent metal ion
; HOMING ENDONUCLEASE
; NUCLEASE DOMAIN
; COLICIN E7
|WOS类目||Biotechnology & Applied Microbiology
Zhang, LK,Huang, YC,Xu DD,et al. Biochemical characterization of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2016,100(18):8003-8012.
Zhang, LK.,Huang, YC.,徐丹丹.,龚勇.,Xu, DD.,...&Ma, KS.(2016).Biochemical characterization of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,100(18),8003-8012.
Zhang, LK,et al."Biochemical characterization of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 100.18(2016):8003-8012.