Abstract
Cell-free extracts of Campylobacter sputorum subspecies bubulus contained superoxide dismutase. The enzyme was located in the cytoplasmic fraction and insensitive to cyanide. After centrifuging a cell-free extract at 144000 x g for 1.5 h the total activity in the supernatant fraction was threefold higher than in the crude cell-free extract. The pellet fraction thus obtained was shown to have a lowering effect on superoxide dismutase activities from different sources in the assay method used here. C. sputorum responded to a raised oxygen tension in the culture by an increase in the superoxide dismutase activity. The ability to produce superoxide anion radicals (O2 -·) during oxidation of formate and lactate was demonstrated. Furthermore C. sputorum was found to produce H2O2 while oxidizing formate. In experiments in which the reduction of cytochrome c by formate was followed, step-wise kinetics were observed. One of the steady states then obtained was attributed to the oxidizing action of H2O2, because it was abolished by the addition of catalase and lengthened by H2O2 added in addition to H2O2 formed as a product of formate oxidation. An overall reaction for formate oxidation by C. sputorum is discussed.
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Abbreviations
- O2 -·:
-
superoxide anion radical
- NBT:
-
p-nitro blue tetrazolium chloride
- ABTS:
-
2,2-azino-di-[3-ethylbenzthiazoline sulfonate (6)]
- TL-medium:
-
tryptose-lactate medium
References
Beauchamp, C., Fridovich, I.: Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analyt. Biochem. 44, 276–287 (1971)
Britton, L., Fridovich, I.: Intracellular localization of the superoxide dismutases of Escherichia coli: a reevaluation. J. Bacteriol. 131, 815–820 (1977)
Carlsson, J., Wrethén, J., Beckman, G.: Superoxide dismutase in Bacteroides fragilis and related Bacteroides species. J. Clin. Microbiol. 6, 280–284 (1977)
Costilow, R. N., Keele, B. B., Jr.: Superoxide dismutase in Bacillus popilliae. J. Bacteriol. 111, 628–630 (1972)
Dailey, H. A., Jr.: Membrane-bound respiratory chain of Spirillum itersonii. J. Bacteriol. 127, 1286–1291 (1976)
Davis, B. J.: Disc gel electrophoresis. Ann. N. Y. Acad. Sci. 121, 404–427 (1964)
Fridovich, I.: Superoxide dismutases. In: Advances in enzymology, Vol. 41 (A. Meister, ed.), pp. 35–97. New York: Wiley 1974
Gregory, E. M., Fridovich, I.: Induction of superoxide dismutase by molecular oxygen. J. Bacteriol. 114, 543–548 (1973)
Gregory, E. M., Kowalski, J. B., Holdeman, L. V.: Production and some properties of catalase and superoxide dismutase from the anaerobe Bacteroides distasonis. J. Bacteriol. 129, 1298–1302 (1977)
Hassan, H. M., Fridovich, I.: Enzymatic defenses against the toxicity of oxygen and of streptonigrin in Escherichia coli. J. Bacteriol. 129, 1574–1583 (1977a)
Hassan, H. M., Fridovich, I.: Regulation of the synthesis of superoxide dismutase in Escherichia coli. Induction by methyl viologen. J. Biol. Chem. 252, 7667–7672 (1977b)
Hatchikian, E. C., Henry, Y. A.: An iron-containing superoxide dismutase from the strict anaerobe Desulfovibrio desulfuricans (Norway 4). Biochimie 59, 153–161 (1977)
Hewitt, J., Morris, J. G.: Superoxide dismutase in some obligately anaerobic bacteria. FEBS-Lett, 50, 315–318 (1975)
Kanematsu, S., Asada, K.: Superoxide dismutase from an anaerobic photosynthetic bacterium, Chromatium vinosum. Arch. Biochem. Biophys. 185, 473–482 (1978)
Koppenol, W. H., Butler, J.: Mechanism of reactions involving singlet oxygen and the superoxide anion. FEBS-Lett. 83, 1–6 (1977)
Loesche, W. J., Gibbons, R. J., Socransky, S. S.: Biochemical characteristics of Vibrio sputorum and relationship to Vibrio bubulus and Vibrio fetus. J. Bacteriol. 89, 1109–1116 (1965)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Muraoka, S., Enomoto, H., Sugiyama, M., Yamasaki, H.: The mechanism of the reduction of cytochrome c by xanthine oxidase. Biochim. Biophys. Acta (Amst.) 143, 408–415 (1967)
Niekus, H. G. D., de Vries, W., Stouthamer, A. H.: The effect of different dissolved oxygen tensions on growth and enzyme activities of Campylobacter sputorum subspecies bubulus. J. Gen. Microbiol. 103, 215–222 (1977)
O'Brien, R. W.: Enzymatic analysis of the pathways of glucose catabolism and gluconeogenesis in Pseudomonas citronellolis. Arch. Microbiol. 103, 71–76 (1975)
Pritchard, G. G., Wimpenny, J. W. J., Morris, H. A., Lewis, M. W. A., Hughes, D. E.: Effects of oxygen on Propionibacterium shermanii grown in continuous culture. J. Gen. Microbiol. 102, 223–233 (1977)
Sinclair, P. R., White, D. C., Smith, L.: Oxidation and reduction of membrane-bound cytochrome c in Hemophilus parainfluenzae. Reaction with oxygen, hydrogen peroxide and nitrate. Biochim. Biophys. Acta (Amst.) 449, 37–47 (1976)
Taylly, F. P., Goldin, B. R., Jacobus, N. V., Gorbach, S. L.: Superoxide dismutase in anaerobic bacteria of clinical significance. Infect. Immun. 16, 20–25 (1977)
Yost, F. J., Jr., Fridovich, I.: Superoxide and hydrogen peroxide in oxygen damage. Arch. Biochem. Biophys. 175, 514–519 (1976)
Yousten, A. A., Bulla, L. A., Jr., McCord, J. M.: Superoxide dismutase in Bacillus popilliae, a catalaseless aerobe. J. Bacteriol. 113, 524–525 (1973)
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Niekus, H.G.D., Wouters, C.H., de Vries, W. et al. Superoxide dismutase and hydrogen peroxide formation in Campylobacter sputorum subspecies bubulus . Arch. Microbiol. 119, 37–42 (1978). https://doi.org/10.1007/BF00407925
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DOI: https://doi.org/10.1007/BF00407925