Abstract
Two forms of iron superoxide dismutase (SOD) were purified from cell extract (CE) and culture filtrate (CF) of Mycobacterium bovis BCG, respectively. The molecular weight of both enzymes was estimated to be approximately 84,000 by gel filtration, whereas that of their subunits was 21,500, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that each of purified enzymes is composed of four identical subunits. The specific activities of CE SOD and CF SOD were 3,850 and 4,040, respectively. The purified enzymes were not joined by disulfide bonds and were, to some extent, resistant to sodium dodecyl sulfate. Their activities were lost by H2O2, but not by azide and cyanide, indicating iron SODs. Enzyme activities were detectable over a broad range of pHs, from 5.0 to 9.0, and were stable for 6 months at -20 degreesC. Each value of pI was 4.5. In Western blots, both enzymes reacted with sera of tuberculosis patients, but not with normal sera. The N-terminal amino acid sequences of CE SOD and CF SOD were the same, suggesting that there is no N-terminal signal sequence.