Biotechnology Letters

, Volume 20, Issue 5, pp 483–488 | Cite as

Cloning and Sequencing of the nrdF Gene of Corynebacterium Ammoniagenes ATCC 6872 Encoding the Functional Metallo-Cofactor of the Manganese-Ribonucleotide Reductase (Mn-RRase)

  • W. Oehlmann
  • U. Griepenburg
  • G. Auling


Cloning of a 4.3 kb DNA fragment from Corynebacterium (formerly Brevibacterium) ammoniagenes ATCC 6872 allowed sequencing and homologous expression of the nrdF gene encoding the metallo-cofactor (CA2 protein) of the Mn-RRase. Due to a multi copy effect transconjugants of C. ammoniagenes showed a sixfold increased resistance towards the radical-scavenger hydroxyurea (HU). The organic radical in the purified CA2 protein generated a visible absorption at 437 nm, absent upon manganese-depletion, and a distinct fluorescence at 513 nm. The two signals were each quenched by HU.


Organic Chemistry Bioorganic Chemistry Hydroxyurea Organic Radical Corynebacterium 
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  1. Auling, G., Blasczyk, K., Griepenburg, U., Hansen, J., Luo, C-H. and Oehlmann, W. (1997). The diversity of metal centers in ribonucleotide reductases. Studies of the bacterial manganese enzyme. In: Bioinorganic Chemistry — Transition Metals in Biology and Their Coordination Chemistry, Trautwein, A.X., ed, pp. 189-203. Research Report-DFG, Wiley-VCH, Weinheim.Google Scholar
  2. Auling, G. and Follmann, H. (1994). Manganese-dependent ribonucleotide reduction and overproduction of nucleotides in coryneform bacteria. In: Metal Ions in Biological Systems. Sigel, H. and A. Sigel eds, vol. 30, pp. 131-164: Metalloenzymes involving amino acid-residues and related radicals, Marcel Dekker Inc., New York.Google Scholar
  3. Auling, G. (1983). Eur. J. Appl. Microbiol. Biotechnol. 18: 229-235.Google Scholar
  4. Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidemann, J.G., Smitz, J.A. and Struhl, K. (1990). Current Protocols in Molecular Biology. Greene Publishing and Wiley Interscience, New York.Google Scholar
  5. Birnboim, H.C. and Doly, J. (1979). Nucleic Acids Res. 7: 1513-1523.PubMedGoogle Scholar
  6. Blattner, F.R., Plunkett III, G., Bloch, C.A., Perna, N.T., Burland, V., Riley, M., Collado-Vides, J., Glasner, J.D., Rode, C.K., Mayhew, G.F., Gregor, J., Davis, N.W., Kirkpatrick, H.A., Goeden, M.A., Rose, D.J., Mau, B., and Shao, Y. (1997). Science 277: 1453-1474.PubMedGoogle Scholar
  7. Follmann, H. (1982). Naturwissenschaften 69: 75-81.PubMedGoogle Scholar
  8. Griepenburg, U., Laßmann, G. and Auling, G. (1996). Free Radical Res. 26: 473-481.Google Scholar
  9. Jordan, A., Gibert, I. and Barbé, J. (1994). J. Bacteriol. 176: 3420-3427.PubMedGoogle Scholar
  10. Luo, C.-H., Hansen, J. and Auling, G. (1997). Arch. Microbiol. 167: 317-324.PubMedGoogle Scholar
  11. Oehlmann, W., Walter, A., Auling, G. (1997). BIOspektrum, abstracts of the VAAM-meeting (ISSN 0947-0867) 16.–19. March, p. 41.Google Scholar
  12. Piepersberg, W. (1993). Streptomycetes and corynebacteria. In: Biotechnology, Rehm, H.J., Reed, G., Pühler, A., Stadler, P., eds, vol. 1, pp. 433-468, VCH, Weinheim.Google Scholar
  13. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.Google Scholar
  14. Schäfer, A., Kalinowski, J., Simon, R., Seep-Feldhaus, A.H. and Pühler, A. (1990). J. Bacteviol. 172: 1663-1666.Google Scholar
  15. Simon, R., Priefer, U. and Pühler, A. (1983). Bio/Technology 1: 784-794.CrossRefGoogle Scholar
  16. Sjöberg, B.-M. (1997). Struct. Bonding 88: 139-173.Google Scholar
  17. Stubbe, J. and van der Donk, W. (1995). Chem. Biol. 2: 793-801.PubMedGoogle Scholar
  18. Willing, A., Follmann, H., Auling, G. (1988). Eur. J. Biochem. 170: 603-611.PubMedGoogle Scholar
  19. Yang, F., Lu, G. and Rubin, H. (1994). J. Bacteriol. 176: 6738-6743.PubMedGoogle Scholar
  20. Yang, F., Curran, S.C., Li, L.-S., Avarbock, D., Graf, J.D., Chua, M-M., Lu, G., Salem, J. and Rubin, H. (1997). J. Bacteriol. 179: 6408-6415.PubMedGoogle Scholar

Copyright information

© Chapman & Hall 1998

Authors and Affiliations

  • W. Oehlmann
    • 1
  • U. Griepenburg
    • 1
  • G. Auling
    • 1
  1. 1.Institut für MikrobiologieUniversität HannoverHannoverGermany

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