Abstract
A superoxide dismutase gene from thermotolerant Bacillus sp. MHS47 (MnSOD47) was cloned, sequenced, and expressed. The gene has an open reading frame of 612 bp, corresponding to 203 deduced amino acids, with high homology to the amino acid sequences of B. thuringiensis (accession no. EEN01322), B. anthracis (accession no. NP_846724), B. cereus (accession no. ZP_04187911), B. weihenstephanensis (accession no. YP_001646918), and B. pseudomycoides. The conserved manganese-binding sites (H28, H83, D165, and H169) show that MnSOD47 has the specific characteristics of the manganese superoxide dismutase (MnSOD) enzymes. MnSOD47 expressed an enzyme with a molecular weight of approximately 22.65 kDa and a specific activity of 3537.75 U/mg. The enzyme is active in the pH range 7–8.5, with an optimum pH of 7.5, and at temperatures in the range 30–45 °C, with an optimum temperature of 37 °C. Tests of inhibitors and metal ions indicated that the enzyme activity is inhibited by sodium azide, but not by hydrogen peroxide or potassium cyanide. These data should benefit future studies of MnSODs in other microorganisms and the biotechnological production of MnSOD47, and could also be used to develop a biosensor for the detection of antioxidants and free radical activity. In the future, this basic knowledge could be applicable to the detection of cancer risks in humans and therapeutic treatments.
Highlights
Superoxide dismutase (SOD) is a metabolic enzyme that catalyzes the conversion of the superoxide radical (O2−) to H2O2 and O2
We focused on the isolation and characterization of the gene encoding SOD in the thermotolerant microorganism Bacillus sp
Ribosomal DNA analysis and this sequence was deposited in GenBank under accession number HQ166833
Summary
Superoxide dismutase (SOD) is a metabolic enzyme that catalyzes the conversion of the superoxide radical (O2−) to H2O2 and O2. SODs are considered key enzymes in the control of oxidative stress because they can protect oxygen-metabolizing cells against the harmful effects of superoxide free radicals [1,2,3,4]. SODs have been used in gene therapy and therapeutic treatments for oxidative damage in the treatment of postischemic reperfusion injury, rheumatoid arthritis and osteoarthritis, brain trauma, influenza-induced lung pneumonitis, breast cancer, nervous system dysfunction, persistent pulmonary hypertension, and tissue damage. A SOD biosensor has been used to determine the antioxidant properties of acetylsalicylic-acid-based drugs and the antiradical activity of healthy and cancerous human brain tissues [18]
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