Cloning and characterization of a new manganese superoxide dismutase from deep-sea thermophile Geobacillus sp. EPT3
Abstract
A new gene encoding a superoxide dismutase (SOD) was identified from a thermophile Geobacillus sp. EPT3 isolated from a deep-sea hydrothermal field in east Pacific. The open reading frame of this gene encoded 437 amino acid residues. It was cloned, overexpressed in Escherichia coli (DE3), and the recombinant protein was purified to homogeneity. Geobacillus sp. EPT3 SOD was of the manganese-containing SOD type, as judged by the insensitivity of the recombinant enzyme to both KCN and H₂O₂, and the activity analysis of Fe or Mn reconstituted SODs by polyacrylamide gel electrophoresis. The recombinant SOD was determined to be a homodimer with monomeric molecular mass of 59.0 kDa. In comparison with other Mn-SODs, the manganese-binding sites are conserved in the sequence (His260, His308, Asp392, His396). The recombinant enzyme had high thermostability at 50 °C. It retained 57 % residual activity after incubation at 90 °C for 1 h, which indicated that this SOD was thermostable. The enzyme also showed striking stability over a wide range of pH 5.0-11.0. At tested conditions, the recombinant SOD from Geobacillus sp. EPT3 showed a relatively good tolerance to some inhibitors, detergents, and denaturants, such as β-mercaptoethanol, dithiothreitol, phenylmethylsulfonyl fluoride, Chaps, Triton X-100, urea, and guanidine hydrochloride.
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SODs can be inhibited by copper chelators such as EDTA and cuprizone.
“SODs can be inhibited also by copper chelators such as EDTA and cuprizone (a copper‐specific chelator) (Zhu et al. 2014).”
From Reactive Oxygen Species, Toxicity, Oxidative Stress, and Antioxidants: Chronic Diseases and Aging (2023) · cites this paper as Zhu et al. 2014
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Reactive Oxygen Species, Toxicity, Oxidative Stress, and Antioxidants: Chronic Diseases and Aging
Jomova, K., Raptova, R., Alomar, S. Y., Alwasel, S. H., Nepovimova, E., Kuca, K., & Valko, M. (2023) · Archives of Toxicology · doi:10.1007/s00204-023-03562-9 · Cited by 2,638 · see citing works
- Authors
- Zhu, Y., Wang, G., Ni, H., Xiao, A., & Cai, H.
- Affiliations
- Jimei University (CN)
- Henan Medical University (CN)
- Corresponding author
- Cai, Huinong
- Venue
- World Journal of Microbiology and Biotechnology
- Published
- 2013-11-17
- Volume
- 30
- Issue
- 4
- Pages
- 1347-1357
- ISSN
- 0959-3993 (ISSN-L)
- 1573-0972
- OpenAlex
- W1989036315
- Cited by
- 41
- References
- 66
- Citations by year
- 2026: 2 · 2025: 3 · 2023: 8 · 2022: 1 · 2021: 3 · 2020: 4 · 2019: 3 · 2018: 4 · 2017: 6 · 2016: 3 · 2015: 3 · 2014: 1
- FWCI
- 2.04
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- World Journal of Microbiology and Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme function and inhibition
- Keywords
- manganese superoxide dismutase, hydrothermal vents, thermal stability, enzyme stability, manganese binding site, recombinant protein expression, protein denaturation
- MeSH terms
- Cloning, Molecular, DNA, Bacterial, Enzyme Inhibitors, Enzyme Stability, Escherichia coli, Hydrogen-Ion Concentration, Molecular Sequence Data, Molecular Weight, Recombinant Proteins, Seawater, Superoxide Dismutase, Temperature, Gene Expression, Open Reading Frames, Conserved Sequence, Sequence Analysis, DNA, Protein Multimerization, Geobacillus
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