Skip to main content
SpringerLink
Log in

Localization and stability of hydrogenases from aerobic hydrogen bacteria

  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Alcaligenes eutrophus strains H 16, B 19, G 27 and N9A contained two different hydrogenases. One enzyme catalyzed the reduction of NAD by hydrogen and was strictly localized in the soluble cell fraction, while the second enzyme was found to be particulate and unable to react with NAD.

All other tested strains, Alcaligenes paradoxus SA 29, Pseudomonas facilis, P. palleronii RH 2, Pseudomonas sp. strain GA 3, Paracoccus denitrificans, Aquaspirillum autotrophicum SA 32, and Corynebacterium autotrophicum 14g and 7C contained only a single enzyme exclusively bound to membranes. This was established using fractional centrifugation, indicator enzyme systems, gentle methods of cell disintegration and discontinuous sucrose density gradient centrifugation. In cell-free extracts obtained by rough disruption (sonication) of cells, hydrogenase was associated to particles of different size and sedimentation velocity. A partial solubilization of hydrogenase caused by sonication was observed with P. facilis.

Without exception, the particulate hydrogenases were found (1) to be unable to reduce pyridine nucleotides, and (2) to reduce methylene blue at an extremely high activity. The eminent reaction rate of 34 μmoles H2 oxidized per min and mg protein has been determined in particle suspensions of Pseudomonas sp. strain GA 3. All hydrogenases were stable during storage under hydrogen atmosphere, except the soluble enzyme from A. eutrophus H 16 which was shown to be more stable under aerobic conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ackrell, B. A. C., Asato, R. N., Mower, H. F.: Multiple forms of bacterial hydrogenases. J. Bact. 92, 828–838 (1966)

    Google Scholar 

  • Aggag, M., Schlegel, H. G.: Studies on a gram-positive hydrogen bacterium, Nocardia opaca strain 1b. I. Description and physiological characterization. Arch. Mikrobiol. 88, 299–318 (1973)

    Google Scholar 

  • Ahrens, J.: Über die Komponenten des Elektronentransportsystems bei Hydrogenomonas H 16. Thesis, Univ. Göttingen (1966)

  • Akagi, J. M., Cambell, L. L.: Studies on thermophilic sulfatereducing bacteria. II. Hydrogenase activity of Desulfotomaculum nigrificans. J. Bact. 82, 927–932 (1961)

    Google Scholar 

  • Aragno, M., Schlegel, H. G.: Aquaspirillum autotrophicum sp. nov., a new hydrogen oxidizing facultatively autotrophic bacterium. Int. J. Syst. Bact. (in press)

  • Aspen, A. J., Wolin, M. J.: Solubilization and reconstitution of a particulate hydrogenase from Vibrio succinogenes. J. biol. Chem. 241, 4152–4156 (1966)

    Google Scholar 

  • Atkinson, D. E., McFadden, B. A.: The biochemistry of Hydrogenomonas. I. The hydrogenase of H. facilis in cell-free preparations. J. biol. Chem. 210, 885–893 (1954)

    Google Scholar 

  • Beisenherz, G., Bolze, H. J., Bücher, T., Czok, R., Garbade, K. H., Meyer-Arendt, E., Pfleiderer, G.: Diphosphofructose-Aldolase, Phosphoglyceraldehyd-Dehydrogenase, Milchsäure-Dehydrogenase, Glyzerophosphat-Dehydrogenase und Pyruvat-Kinase aus Kaninchenmuskulatur in einem Arbeitsgang. Z. Naturforsch. 8b, 555–577 (1953)

    Google Scholar 

  • Bell, G. R., LeGall, J., Peck, H. D., Jr.: Evidence for the periplasmic location of hydrogenase in Desulfovibrio gigas. J. Bact. 120, 994–997 (1974)

    Google Scholar 

  • Bergmeyer, H. U.: Zur Messung von Katalase-Aktivitäten. Biochem. Z. 327, 255–258 (1955)

    Google Scholar 

  • Bone, D. H.: Localization of H2 activity enzymes in Pseudomonas saccharophila. Biochem. biophys. Res. Commun. 3, 211–214 (1960)

    Google Scholar 

  • Buller, C. S., Akagi, J. M.: Hydrogenase of Coleman's sulfatereducing bacterium. J. Bact. 88, 440–443 (1964)

    Google Scholar 

  • Burns, R. C., Bulen, W. A.: ATP-dependent hydrogen evolution by cell-free preparations of Azotobacter vinelandii. Biochim. biophys. Acta (Amst.) 105, 437–445 (1965)

    Google Scholar 

  • Canevascini, G., Eberhardt, U.: Chemolithotrophic growth and regulation of hydrogenase formation in the coryneform hydrogen bacterium strain 11/x. Arch. Microbiol. 103, 283–291 (1975)

    Google Scholar 

  • Eberhardt, U.: Über das wasserstoffaktivierende System bei Hydrogenomonas H 16. I. Verteilung der Hydrogenase-Aktivität auf zwei Zellfraktionen. Arch. Mikrobiol. 53, 288–302 (1966)

    Google Scholar 

  • Eberhardt, U.: On chemolithotrophy and hydrogenase of a grampositive Knallgasbacterium. Arch. Mikrobiol. 66, 91–104 (1969)

    Google Scholar 

  • Eberhardt, U.: The cell wall as the site of carotenoid in the “Knallgas”-bacterium, 12/60/x. Arch. Mikrobiol. 80, 32–37 (1971)

    Google Scholar 

  • Findley, J. E., Akagi, J. M.: Lysis of Desulfovibrio vulgaris by ethylene diaminetetraacetic acid and lysozyme. J. Bact. 96, 1427–1428 (1968)

    Google Scholar 

  • Gogotov, J. N., Zorin, N. A., Bogorov, L. V.: Hydrogen metabolism and ability for nitrogen fixation in Thiocapsa roseopersicina. Microbiology 43, 1–6 (1974)

    Google Scholar 

  • Haschke, R. H., Campbell, L. L.: Purification and properties of a hydrogenase from Desulfovibrio vulgaris. J. Bact. 105, 249–258 (1971)

    Google Scholar 

  • Klemme, J. H.: Studies on the mechanism of NAD-photoreduction by chromatophores of the facultative phototroph Rhodopseudomonas capsulata. Z. Naturforsch. 24b, 67–76 (1969)

    Google Scholar 

  • Klemme, J. H.: Alosterisch regulierte und nicht regulierte Pyrophosphat-Phosphohydrolasen aus phototrophen Bakterien. Inauguraldissertation, Univ. Göttingen (1972)

  • Kidman, A. D., Yanagihara, R., Asato, R. N.: Comparative studies of bacterial hydrogenase. Biochim. biophys. Acta (Amst.) 191, 170–173 (1969)

    Google Scholar 

  • Krasna, A. J., Rittenberg, D.: The hydrogenase of Proteus vulgaris. Biochim. biophys. Acta (Amst.) 26, 526–530 (1957)

    Google Scholar 

  • Kühnemund, H.: Zur Verwertung von molekularem Wasserstoff durch Micrococcus denitrificans. Thesis, Univ. Göttingen (1971)

  • LeGall, J., Dervartanian, D. V., Spilker, E., Lee, J. P., Peck, H. D., Jr.: Evidence for the involvement of non-heme iron in the active site of hydrogenase from Desulfovibrio vulgaris. Biochim. biophys. Acta (Amst.) 234, 525–530 (1971)

    Google Scholar 

  • Nakos, G., Mortenson, L. E.: Structural properties of hydrogenase from Clostridium pasteurianum W5. Biochemistry 10, 2442–2449 (1971)

    Google Scholar 

  • Peck, H. D., Jr., Gest, H.: Fumarate reduction by molecular hydrogen in cell-free systems. Bact. Proc. 54, 122 (1954)

    Google Scholar 

  • Probst, I., Schlegel, H. G.: Respiratory components and oxidase activities in Alcaligenes eutrophus. Biochim. biophys. Acta (Amst.) 440, 412–418 (1976)

    Google Scholar 

  • Racker, E.: Resolution and reconstitution of the inner mitochondrial membrane. Fed. Proc. 26, 1335–1340 (1967)

    Google Scholar 

  • Reddy, C. A., Bryant, M. P., Wolin, M. J.: Ferredoxin-dependent conversion of acetaldehyde to acetate and H2 in extracts of S organism. J. Bact. 110, 133–138 (1972)

    Google Scholar 

  • Schlegel, H. G., Kaltwasser, H., Gottschalk, G.: in Submersverfahren zur Kultur wasserstoffoxydierender Bakterien: Wachstumsphysiologische Untersuchungen. Arch. Mikrobiol. 38, 209–222 (1961)

    Google Scholar 

  • Schneider, K., Rudolph, V., Schlegel, H. G.: Description and physiological characterization of a coryneform hydrogen bacterium, strain 14g. Arch. Mikrobiol. 93, 179–193 (1973)

    Google Scholar 

  • Schneider, K., Schlegel, H. G.: Purification and properties of soluble hydrogenase from Alcaligenes eutrophus H 16. Biochim. biophys. Acta (Amst.) 452, 66–80 (1976)

    Google Scholar 

  • Temperli, A., Pengra, R. M., Wilson, P. W.: Some properties of a soluble and particle-bound hydrogenase in Aerobacter aerogenes. Biochim. biophys. Acta (Amst.) 38, 557–558 (1960)

    Google Scholar 

  • Tunail, N., Schlegel, H. G.: A new coryneform hydrogen bacterium: Corynebacterium autotrophicum strain 7C. I. Characterization of the wild type strain. Arch. Microbiol. 100, 341–350 (1974)

    Google Scholar 

  • Vishniac, W., Trudinger, P. A.: Carbon dioxide fixation and substrate oxidation in chemosynthethic sulfur and hydrogen bacteria. In: Symposium on autotrophy. Bact. Rev. 26, 168–169 (1962)

  • Wittenberger, H., Repaske, R.: Studies on hydrogen in cell-free extracts of Hydrogenomonas eutropha. Biochim. biophys. Acta (Amst.) 47, 542–552 (1961)

    Google Scholar 

  • Yagi, T.: Solubilization, purification and properties of particulate hydrogenase from Desulfovibrio vulgaris. J. Biochem. 68, 649–657 (1970)

    Google Scholar 

  • Yagi, T., Honya, M., Tamiya, N.: Purification and properties of hydrogenases of different origins. Biochim. biophys. Acta (Amst.) 153, 699–705 (1968)

    Google Scholar 

  • Yagi, T., Kimura, K., Daidoji, H., Sakai, F., Tamura, S., Inokuchi, H.: Properties of purified hydrogenase from the particulate fraction of Desulfovibrio vulgaris, Miyazaki. J. biol. Chem. 79, 661–671 (1976)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie der Gesellschaft für Strahlen- und Umweltforschung mbH München, Grisebachstr. 8, D-3400, Göttingen, Germany

    Klaus Schneider & Hans G. Schlegel

  2. Institut für Mikrobiologie der Universität Göttingen, Grisebachstr. 8, D-3400, Göttingen, Germany

    Klaus Schneider & Hans G. Schlegel

Authors
  1. Klaus Schneider
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hans G. Schlegel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schneider, K., Schlegel, H.G. Localization and stability of hydrogenases from aerobic hydrogen bacteria. Arch. Microbiol. 112, 229–238 (1977). https://doi.org/10.1007/BF00413086

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00413086

Key words

Search

Navigation