Summary
Bacillus halodenitrificans produced a dimeric, manganese-containing superoxide dismutase constitutively when grown either aerobically or as a denitrifier. The molecular mass of the enzyme was determined by sedimentation equilibrium to be 41.4±3 kDa with each subunit estimated at 26 kDa. Plasma emission spectroscopy indicated the presence of 1.22 mol manganese atoms/mol holoenzyme. The electronic absorption spectrum displayed a broad band centered at approximately 474 nm (ε=560 mM−1 · cm−1) and a shoulder at 595 nm. In the ultraviolet range, the spectrum exhibited split maxima at 278 nm and 283 nm and a shoulder at 291 nm, thus resembling the spectra of superoxide dismutase fromBacillus subtilis andEscherichia coli. The amino acid composition of theB. halodenitrificans enzyme differed slightly quantitatively but little qualitatively from counterpart enzymes from other sources. Like the superoxide dismutases ofMycobacterium lepraemurium and human mitochondria, theB. halodenitrificans enzyme exhibited several cysteine residues. As expected from the capacity superoxide dismutase exhibits for protecting NO as neutrophil cytotoxicity factor, theB. halodenitrificans superoxide dismutase did not interfere with accumulation of NO produced by the organism's nitrite reductase.
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References
Bannister JV, Rotilio G (1984) A decade of superoxide dismutase activity. Dev Biochem 26:146–185
Barra D, Schinina ME, Simmaco M, Bannister JV, Bannister WH, Rotilio G, Bossa F (1984) The primary structure of human liver manganese superoxide dismutase. J Biol Chem 259:12595–12601
Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287
Britton L, Malinowski D, Fridovich I (1978) Superoxide dismutase and oxygen metabolism inStreptococcus faecalis and comparisons with other organisms. J Bacteriol 134:229–236
Brock CJ, Walker JE (1980) Superoxide dismutase fromBacillus stearothermophilus. Complete amino acid sequence of a manganese enzyme. Biochemistry 19:2873–2882
Carlioz A, Touati D (1986) Isolation of superoxide dismutase mutants inEscherichia coli: is superoxide dismutase necessary for life? EMBO J 5:623–630
DiGuiseppi J, Fridovich I (1982) Oxygen toxicity inStreptococcus sanguis. The relative importance of superoxide and hydroxyl radicals. J Biol Chem 257:4046–4051
Epp J, Fairchild S, Erickson G, Koppenol WH (1986) Superoxide dismutase activity of manganese chelates. In: Rotilio G (ed) Superoxide and superoxide dismutase in chemistry, biology and medicine. Elsevier, Amsterdam, pp 76–78
Fee JA (1981) Is superoxide toxic and are superoxide dismutases essential for aerobic life? In: Rodgers MAJ, Powers EL (eds) Oxygen and oxyradicals in chemistry and biology, Academic Press, London, pp 205–239
Gregory EM (1985) Characterization of the manganese-containing superoxide dismutase fromBacteroides fragilis. Arch Biochem Biophys 238:83–89
Gregory EM, Dapper CH (1980) Chemical and physical differentiation of superoxide dismutases in anaerobes. J Bacteriol 144:967–974
Gregory EM, Dapper CH (1983) Isolation of iron-containing superoxide dismutase fromBacteroides fragilis: reconstitution as a Mn-containing enzyme. Arch Biochem Biophys 220:293–300
Gregory EM, Fridovich I (1973) Induction of superoxide dismutase by molecular oxygen. J Bacteriol 114:543–548
Hassan HM, Moody CS (1986) Regulation of the biosynthesis of superoxide dismutase in procaryotes. In: Ratilio G (ed) Superoxide and superoxide dismutase in chemistry, biology and medicine. Elsevier, Amsterdam, pp 264–279
Hatchikian EC, LeGall J, Bell GR (1977) Significance of superoxide dismutase and catalase activities in the strict anaerobes, sulfate-reducing bacteria. In: Michelson AM, McCord JM, Fridovich I (eds) Superoxide and superoxide dismutase. Academic Press, London, pp 159–172
Henry Y, Bessieres P (1984) Denitrification and nitrite reduction:Pseudomonas aeruginosa nitrite reductase. Biochimie 66:259–289
Ichihara K, Kusunose E, Kusunose M, Mori J (1977) Superoxide dismutase fromMycobacterium lepremurium. J Biochem (Tokyo) 81:1427–1433
Jovin T, Chrambach A, Naughton MA (1964) An apparatus for preparative temperature-regulated polyacrylamide gel electrophoresis. Anal Biochem 9:351–369
Kanematsu S, Asada K (1978) Crystalline ferric superoxide dismutase from an anaerobic green sulfur bacterium,Chlorobium thiosulfatophilum. FEBS Lett 91:94–98
Keele BB Jr, McCord JM, Fridovich I (1970) Superoxide dismutase fromEscherichia coli: a new manganese-containing enzyme. J Biol Chem 245:6176–6181
Kirby TW, Blum J, Kahane I, Fridovich I (1980) Distinguishing between Mn-containing and Fe-containing superoxide dismutases in crude extracts of cells. Arch Biochem Biophys 201:551–555
Kirby TW, Lancaster JR Jr, Fridovich I (1981) Isolation and characterization of the iron-containing superoxide dismutase ofMethanosarcina bryantii. Arch Biochem Biophys 210:140–148
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymatic function for erythrocuprein (hemocuprein). J Biol Chem 244:6049–6055
Meier B, Barra D, Bossa F, Calabrese L, Rotilio G (1982) Synthesis of either Fe- or Mn-superoxide dismutase with an apparently identical protein moiety by an anaerobic bacterium dependent on the metal supplied. J Biol Chem 257:13977–13980
Misra HP, Fridovich I (1978) Inhibition of superoxide dismutase by azide. Arch Biochem Biophys 189:317–322
Miyake K (1986) Effect of nitrate on the level of superoxide dismutase in anaerobically grownEscherichia coli. J Gen Appl Microbiol 32:527–533
Parker MW, Bossa F, Barra D, Bannister WH, Bannister JV (1986) Structural and evolutionary relationships between the eukaryotic superoxide dismutases. In: Rotilio G (ed) Superoxide and superoxide dismutase in chemistry, biology and medicine. Elsevier, Amsterdam, pp 237–245
Pennington CD, Gregory E (1986) Isolation and reconstitution of iron- and manganese-containing superoxide dismutases fromBacteroides thetaiotaomicron. J Bacteriol 166:528–532
Puget K, Michelson AM (1974) Isolation of a new copper-containing superoxide dismutase bacteriocuprein. Biochem Biophys Res Commun 58:830–838
Ravindranath SD, Fridovich I (1975) Isolation and characterization of a manganese-containing superoxide dismutase from yeast. J Biol Chem 250:6107–6112
Read SM, Northcote DH (1981) Minimization of variation in response to different proteins of the Coomassie blue G dyebinding assay for protein. Anal Biocem 116:53–64
Salin ML, Oesterhelt D (1988) Purification of a manganese-containing superoxide dismutase fromHalobacterium halobium. Arch Biochem Biophys 260:806–810
Schachman HK (1959) Ultracentrifugation in biochemistry. Academic Press, New York
Steinman HM (1982) Copper-zinc superoxide dismutase fromCaulobacter crescentus CB15. A novel bacteriocuprein form of the enzyme. J Biol Chem 257:10283–10293
Tsukuda K, Kido T, Shimasue Y, Soda K (1983) Isolation of manganese-containing superoxide dismutase fromBacillus subtilis. Agric Biol Chem 47:2865–2870
Vignais PM, Henry M-F, Terech A, Chabert J (1980) Production of superoxide anion and superoxide chemical and biochemical aspects of superoxide and superoxide dismutase. Dev Biochem 11A:154–159
Wright CD, Mülsch A, Busse R, Osswald H (1989) Generation of nitric oxide by human neutrophils. Biochem Biophys Res Commun 160:813–819
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Denariaz, G., Payne, W.J. & LeGall, J. Characterization of the superoxide dismutase of the denitrifying bacterium,Bacillus halodenitrificans . Biol Metals 3, 14–18 (1990). https://doi.org/10.1007/BF01141171
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DOI: https://doi.org/10.1007/BF01141171