Applied Microbiology and Biotechnology

, Volume 24, Issue 2, pp 106–112 | Cite as

Effect of different limitations in chemostat cultures on growth and production of exocellular protease byBacillus licheniformis

  • Jurjen Frankena
  • Gregory M. Koningstein
  • Henk W. van Verseveld
  • Adriaan H. Stouthamer


Maximal molar growth yields (Y sub max ) and protease production ofBacillus licheniformis S 1684 during NH 4 + -, O2-, and NH 4 + +O2-limitation with either glucose or citrate as carbon and energy source and during glucose-, and citratelimitation in chemostat cultures were determined. Protease production was repressed by excess ammonia when glucose served as C/E-source. Glucose and citrate repressed protease production during NH 4 + -limitation. A low oxygen tension enbanced protease production at low μ-values. It was concluded that, besides ammonia repression, catabolite flux and oxygen tension influence protease production, indicating that the energy status of the cell is important for the level of protease production.Y sub max -values were high during glucose-limitation and indicate a high efficiency of growth caused by a highY ATP max . During NH 4 + -, O2-, and NH 4 + +O2-limitation with glucose as C/E-values were lower than during glucose limitation. The lowerY sub max -values were due to a lower efficiency of energy conservation.Y sub max -values during limitations with citrate as C/E-source were lower than during limitations with glucose as C/E-source.



specific growth rate (h-1)


growth yield per mol substrate (g biomass/mol)


maximal molar growth yield corrected for maintenance requirements (g biomass/mol)

Ymax (corr)

Ymax corrected for product formation (g biomass/mol)


maintenance requirements (mol/g biomass·h)

msub (corr)

maintenance requirements corrected for product formation (mol/g biomass·h)


maximal specific rate of protease production (E440/mg DW·h)


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  1. Bernt E, Gutman I (1970) Äthanol: Bestimmung mit Alkohol-Dehydrogenase und NAD. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Band II. Verlag Chemie, Weinheim/Bergstr., pp. 1457–1460Google Scholar
  2. Dawes EA, McGill DJ, Midgley M (1971) Analysis of fermentation products. In: Norris JR, Ribbons DW (eds) Methods in microbiology, Vol 6A, Academic Press, New York London, pp 53–215Google Scholar
  3. Frankena J, Verseveld HW van, Stouthamer AH (1985) A continuous culture study of the bioenergetic aspects of growth and production of exocellular protease inBacillus licheniformis. Appl Microbiol Biotechnol 22:169–176Google Scholar
  4. Hanlon GW, Hodges NA, Russel AD (1982) The influence of glucose, ammonium and magnesium availability on the production of protease and bacitracin byBacillus licheniformis. J Gen Microbiol 128:845–851Google Scholar
  5. Hohorst HJ (1970)l-lactat: Bestimmung mit Lactat-Dehydrogenase und NAD: In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Band II. Verlag Chemie, Weinheim/Bergstr., pp 1425–1429Google Scholar
  6. Holz G, Bergmeyer HU (1970) Acetat: Bestimmung mit Acetatkinase und Hydroxylamin. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Band II. Verlag Chemie, Weinheim/Bergstr., pp 1487–1490Google Scholar
  7. Laishley EJ, Bernlohr RW (1968) Regulation of arginine and proline catabolism inBacillus licheniformis. J Bacteriol 96:322–329Google Scholar
  8. Lang E, Lang H (1972) Spezifische Farbreaktion zum direkten Nachweise der Ameisensäure. Z Anal Chem 260:8–10Google Scholar
  9. Neijssel OM, Tempest DW (1975) The regulation of carbohydrate metabolism inKlebsiella aerogenes NCTC 418 organisms, growing in chemostat culture. Arch Microbiol 106:251–258Google Scholar
  10. Neijssel OM, Tempest DW (1976) Bioenergetic aspects of aerobic growth ofKlebsiella aerogenes NCTC 418 in carbon-limited and carbon-sufficient chemostat culture. Arch Microbiol 107:215–221Google Scholar
  11. Neijssel OM, Tempest DW (1979) the physiology of metabolite over-production. Symp Soc Gen Microbiol 29:53–82Google Scholar
  12. Pirt SJ (1965) The maintenance energy of bacteria in growing cultures. Proc Roy Soc B 163:224–231Google Scholar
  13. Priest FG (1977) Extracellular enzyme synthesis in the genusBacillus. Bacteriol Rev 41:711–753Google Scholar
  14. Schaeffer P (1969) Sporulation and the production of antibiotics, exoenzymes, and exotoxins. Bacteriol Rev 33:48–71Google Scholar
  15. Stouthamer AH, Bettenhaussen CW (1975) Determination of the efficiency of oxidative phosphorylation in continuous cultures ofAerobacter aerogenes. Arch Microbiol 102:187–192Google Scholar
  16. Stouthamer AH (1979) Energy production, growth and product formation by microorganisms. In: Sebek OK, Laskin AL (eds) Genetics of industrial microorganisms. Am Soc Microbiol, Washington DC pp 70–76Google Scholar
  17. Vries W de, Stouthamer AH (1968) Fermentation of glucose, lactose, galactose, mannitol and xylose byBifidobacteria. J Bacteriol 96:472–478Google Scholar
  18. Whooley MA, O'Callaghan JA, McLoughlin AJ (1983) Effect of substrate on the regulation of exoprotease production byPseudomonas aeruginosa ATCC 10145. J Gen Microbiol 129:981–988Google Scholar
  19. Wiersma M, Harder W (1978) A continuous culture study of the regulation of extracellular protease production inVibrio SA1. Antonie van Leeuwenhoek 44:141–155Google Scholar
  20. Wouters JTM, Buysman PJ (1977) Production of some exocellular enzymes byBacillus licheniformis 749/C in chemostat cultures. FEMS Letters 1:109–112Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Jurjen Frankena
    • 1
  • Gregory M. Koningstein
    • 1
  • Henk W. van Verseveld
    • 1
  • Adriaan H. Stouthamer
    • 1
  1. 1.Department of Microbiology, Biological LaboratoryVrije UniversiteitAmsterdamThe Netherlands

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