Advertisement

Biotechnology Letters

, Volume 25, Issue 2, pp 143–147 | Cite as

Arrest of cell cycle by inhibition of ribonucleotide reductase induces accumulation of NAD+ by Mn2+-supplemented growth of Corynebacterium ammoniagenes

  • Bouziane Abbouni
  • Hesham M. Elhariry
  • Georg AulingEmail author
Article

Abstract

Cell division of the wild type strain Corynebacterium (formerly Brevibacterium) ammoniagenes ATCC 6872 which requires 1 μM Mn2+ for balanced growth was inhibited by addition of 20 mM hydroxyurea (HU) or 10 mM p-methoxyphenol (MP) to a Mn2+-supplemented fermentation medium at an appropriate time. Scanning electron microscopy (SEM) showed a restricted elongation characteristic of arrest of the cell cycle in coryneform bacteria. The cultures treated with HU or MP had, respectively, a fourfold or sixfold enhanced accumulation of NAD+ by a salvage biosynthetic pathway. An assay of nucleotide-permeable cells for ribonucleotide reductase activity using [3H-CDP] as substrate revealed a pre-early and complete decline of DNA precursor biosynthesis not found in the untreated control. Overproduction of NAD+ is an alternative to the conventional fermentation process using Mn2+ deficiency. A simple model is presented to discuss the metabolic regulation of the new process based on the presence of a manganese ribonucleotide reductase (Mn-RNR) in the producing strain.

cell cycle,Corynebacterium ammoniagenes elongation hydroxyurea manganese-ribonucleotide reductase NAD+ p-methoxyphenol scanning electron microscopy 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Auling G, Follmann H (1994) Manganese-dependent ribonucleotide reduction and overproduction of nucleotides in coryneform bacteria. In: Sigel H, Sigel A, eds., Metal Ions in Biological Systems, Vol. 30. Metalloenzymes Involving Amino Acid-Residues and Related Radicals. New York: Marcel Dekker Inc, pp. 131–164.Google Scholar
  2. Chabes A, Thelander L (2000) Controlled protein degradation regulates ribonucleotide reductase activity in proliferating mammalian cells during the normal cell cycle and in response to DNA damage and replication blocks. J. Biol. Chem. 275: 17747–17753.Google Scholar
  3. Griepenburg U, Blasczyk K, Kappl R, Hüttermann J, Auling G (1998) A divalent metal site in the small subunit of manganesedependent ribonucleotide reductase of Corynebacterium ammoniagenes. Biochemistry 37: 7992–7996.Google Scholar
  4. Kijima N, Goyal D, Takada A, Wachi M (1998) Induction of only limited elongation instead of filamentation by inhibition of cell division in Corynebacterium glutamicum. Appl. Microbiol. Biotechnol. 50: 227–232.Google Scholar
  5. Liebl W, Klamer R, Schleifer K (1989) Requirement of chelating compounds for the growth of Corynebacterium glutamicum in synthetic media. Appl. Microbiol. Biotechnol. 32: 205–210.Google Scholar
  6. Luo C-H, Hansen J, Auling G (1997) Temperature-sensitive mutants of Corynebacterium ammoniagenes ATCC 6872 with a defective large subunit of the manganese-containing ribonucleotide reductase. Arch. Microbiol. 167: 317–324.Google Scholar
  7. Mihara Y, Utagawa T, Yamada H, Asano Y (2000) Phosphorylation of nucleosides by a mutated acid phosphatase from Morganella morganii. Appl. Environ. Microbiol. 66: 2811–2816.Google Scholar
  8. -monophosphate production using overexpressed guanosine/ inosine kinase. Appl. Microbiol. Biotechnol. 48: 693–698.Google Scholar
  9. Nagodawithana TW (1994) Savory flavors. In: Gabelman A, ed., Bioprocess Production of Flavor, Fragrance, and Color Ingredients. New York: John Wiley &; Sons Inc., pp. 135–168.Google Scholar
  10. Nakayama K, Sato Z, Tanaka H, Kinoshita S (1968) Production of nucleic acid-related substances by fermentative processes. Part XVII. Production of NAD and nicotinic acid mononucleotide with Brevibacterium ammoniagenes. Agric. Biol. Chem. 32: 1331–1336.Google Scholar
  11. Nara T, Misawa M, Komuro T, Kinoshita S (1969) Production of nucleic acid related substances by fermentative processes. Part XXX. Effect of antibiotics and surface active agents on 5′-purine-nucleotide production by Brevibacterium ammoniagenes. Agric. Biol. Chem. 33: 1198–1204.Google Scholar
  12. Satta G, Fontana R, Canepari P (1994) The two-competing site (TCS) model for cell shape regulation in bacteria: the envelope as an integration point for the regulatory circuits of essential physiological events. Adv. Microb. Physiol. 36: 181–245.Google Scholar
  13. Teshiba S, Furuya A (1989) Production of Nucleotides and Nucleosides by Fermentation. Amsterdam: Gordon and Breach Science Publishers, OPA BV.Google Scholar
  14. White D (2000) The Physiology and Biochemistry of Prokaryotes, 2nd edn. New York: Oxford University Press.Google Scholar
  15. Zellner G, Geveke M, De Macario EC, Diekmann H (1991) Population dynamics of biofilm development during start-up of a butyrate-degrading fluidized-bed reactor. Appl. Microb. Biotechnol. 36: 404–409.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Bouziane Abbouni
    • 1
  • Hesham M. Elhariry
    • 1
  • Georg Auling
    • 1
    Email author
  1. 1.Institut für MikrobiologieUniversität HannoverHannoverGermany

Personalised recommendations