Archives of Microbiology

, Volume 157, Issue 2, pp 135–140 | Cite as

Phosphate-limited growth of Chromatium vinosum in continuous culture

  • Jordi Mas
  • Hans van Gemerden
Original Papers


Chromatium vinosum DSM 185 was grown in continuous culture at a constant dilution rate of 0.071 h-1 with sulfide as the only electron donor. The organism was subjected to conditions ranging from phosphate limitation (SR-phosphate=2.7 μM and SR-sulfide=1.8 mM) to sulfide limitation (SR-phosphate=86 μM and SR-sulfide=1.8 mM). At values of SR-phosphate below 7.5 μM the culture was washed out, whereas SR-phosphate above this value resulted in steady states. The saturation constant (Kμ) for growth on phosphate was estimated to be between 2.6 and 4.1 μM. The specific phosphorus content of the cells increased from 0.30 to 0.85 μmol P mg-1 protein with increasing SR-phosphate. The specific rate of phosphate uptake increased with increasing SR-phosphate, and displayed a non-hyperbolic saturation relationship with respect to the concentration of phosphate in the inflowing medium. Approximation of a hyperbolic saturation function yielded a maximum uptake rate (Vmax) of 85 nmol P mg-1 protein h-1, and a saturation constant for uptake (Kt) of 0.7 μM. When phosphate was supplied in excess 8.5% of the phosphate taken up by the cells was excreted as organic phosphorus at a specific rate of 8 nmol P mg-1 protein h-1.

Key words

Chromatium vinosum Phosphate Nutrient limitation Continuous culture Phototrophic bacteria 

Non-standard abbreviations


bacteriochlorophyll a


dilution rate; μmax, maximum specific growth rate


maximum specific growth rate if the substrate were not inhibitory


saturation constant for growth on phosphate


maximum rate of phosphate uptake


saturation constant for phosphate uptake


inhibition constant for growth in the presence of sulfide


concentration of substrate in the inflowing medium


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bader FB (1982) Kinetics of double-substrate limited growth. In: Bazin MJ (ed) Microbial population dynamics. CRC Press, Boca Raton, pp 1–32Google Scholar
  2. Beeftink HH, Gemerden H van (1979) Actual and potential rates of substrate oxidation and product formation in continuous cultures of Chromatium vinosum. Arch Microbiol 121: 161–167CrossRefGoogle Scholar
  3. Bergstein T, Henis Y, Cavari BZ (1979) Investigations on the photosynthetic sulfur bacterium Chlorobium phaeobacteroides causing seasonal blooms in Lake Kinneret. Can J Microbiol 25: 999–1007CrossRefGoogle Scholar
  4. Burns DJW, Beever RE (1977) Kinetic characterization of the two phosphate uptake systems in the fungus Neurospora crassa. J Bacteriol 132: 511–519PubMedPubMedCentralGoogle Scholar
  5. Button DK (1983) Differences between the kinetics of nutrient uptake by micro-organisms, growth and enzyme kinetics. Trends Biochem Sci 8: 121–124CrossRefGoogle Scholar
  6. Button DK (1985) Kinetics of nutrient limited transport and microbial growth. Microb Rev 49: 270–297Google Scholar
  7. deJonge VN, Villerius LA (1980) Interference of sulfide in inorganic phosphate determination in natural waters. Marine Chem 9: 191–197CrossRefGoogle Scholar
  8. deWit R, Gemerden H van (1987) Chemolithotrophic growth of the phototrophic sulfur bacterium Thiocapsa roseopersicina. FEMS Microbiol Ecol 45: 117–126CrossRefGoogle Scholar
  9. Edwards VH (1970) The influence of high substrate concentrations on microbial kinetics. Biotechnol Bioeng 12: 679–712CrossRefGoogle Scholar
  10. Ellwood DC, Tempest DW (1972) Effects of environment on bacterial wall content and composition. Adv Microbial Physiol 7: 83–117CrossRefGoogle Scholar
  11. Gemerden H van, Beeftink HH (1978) Specific rates of substrate oxidation and product formation in autotrophically growing Chromatium vinosum cultures. Arch Microbiol 119: 135–143CrossRefGoogle Scholar
  12. Gemerden H van (1980) Survival of Chromatium vinosum at low light intensities. Arch Microbiol 125: 115–121CrossRefGoogle Scholar
  13. Gemerden H van (1984) The sulfide affinity of phototrophic bacteria in relation to the location of elemental sulfur. Arch Microbiol. 139: 289–294CrossRefGoogle Scholar
  14. Gogotov IN, Glinskii VP (1973) A comparative study of nitrogen fixation in the purple bacteria. Microbiology 42: 877–880Google Scholar
  15. Grillo JF, Gibson J (1979) Regulation of phosphate accumulation in the unicellular cyanobacterium Synechococcus. J Bacteriol 140: 508–517PubMedPubMedCentralGoogle Scholar
  16. Guerrero R, Montesinos E, Pedrós-Alió C, Esteve I, Mas J, Gemerden H van, Hofman PAG, Bakker JF (1985) Phototrophic sulfur bacteria in two Spanish lakes: vertical distribution and limiting factors. Limnol Oceanogr 30: 919–931CrossRefGoogle Scholar
  17. Guerrero R, Pedrós-Alió C, Esteve I, Mas J (1987) Communities of phototrophic sulfur bacteria in lakes of the Spanish Mediterranean region. Acta Academiae Aboensis 47: 125–151Google Scholar
  18. Heda GD, Madigan MT (1986) Aspects of nitrogen fixation in Chlorobium. Arch Microbiol 143: 330–336CrossRefGoogle Scholar
  19. Herbert D, Elsworth R, Telling RC (1956) The continuous culture of bacteria: a theoretical and experimental study. J Gen Microbiol 14: 601–622CrossRefGoogle Scholar
  20. Keppen OI, Lebedeva NV, Petukhov SA, Rodionov YV (1985) Nitrogenase activity in the green bacterium Chlorobium limicola. Microbiology 54: 28–32Google Scholar
  21. Koroleff F (1983) Determination of nutrients. In: Grasshoff K, Erhardt M, Kremling K (eds) Methods of seawater analysis, 2nd edn. Verlag Chemie, Weinheim, pp 125–187Google Scholar
  22. Lowry OH, Rosebrough NH, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275Google Scholar
  23. Maaløe OG, Kjeldgaard NO (1966) Control of macromolecular synthesis. WA Benjamin, New YorkGoogle Scholar
  24. Mas J, Gemerden H van (1987) Influence of sulfur accumulation and composition of the sulfur globule on cell volume and buoyant density of Chromatium vinosum. Arch Microbiol 146: 362–369CrossRefGoogle Scholar
  25. Megee III RD, Drake JF, Fredrickson AG, Tsuchiya HM (1972) Studies in intermicrobial symbiosis. Saccharomyces cerevisiae and Lactobacillus casei. Can J Microbiol 18: 1733–1742CrossRefGoogle Scholar
  26. Minnikin DE, Abdolrahimzadeh H (1974) The replacement of phosphatidylethanolamine and acidic phospholipids by an ornithine-amide lipid and a minor phosphorus-free lipid in Pseudomonas fluorescens NCMB 129. FEBS Lett 43: 257–260CrossRefGoogle Scholar
  27. Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta 27: 31–36CrossRefGoogle Scholar
  28. Ormerod JG, Ormerod KS, Gest H (1961) Light-dependent utilization of organic compounds and photoproduction of molecular hydrogen by photosynthetic bacteria; relationships with nitrogen metabolism. Arch Biochem Biophys 94: 449–463CrossRefGoogle Scholar
  29. Pachmayr F (1960) Vorkommen und Bestimmung von Schwefelverbindungen in Mineralwasser. Thesis, University of Munich, Faculty of Life SciencesGoogle Scholar
  30. Parkin TB, Brock TD (1980) Photosynthetic bacterial production in lakes: the effects of light intensity. Limnol Oceanogr 25: 711–718CrossRefGoogle Scholar
  31. Rhee GY (1973) A continuous culture study of phosphate uptake, growth rate and polyphosphate in Scenedesmus sp. J Phycol 9: 495–506Google Scholar
  32. Riegman R (1985) Phosphate-phytoplankton interactions. PhD Thesis, University of Amsterdam, Faculty of Mathematics and Life SciencesGoogle Scholar
  33. Robertson BR, Button DK (1979) Phosphate-limited continuous culture of Rhodotorula rubra: kinetics of transport, leakage and growth. J Bacteriol 138: 884–895PubMedPubMedCentralGoogle Scholar
  34. Standard methods for the examination of water and wastewater, 16th edn (1985) Am Publ Health Assoc, WashingtonGoogle Scholar
  35. Stal LJ, Gemerden H van, Krumbein WE (1984) The simultaneous assay of chlorophyll and bacteriochlorophyll in natural microbial communities. J Microbiol Methods 2: 295–306CrossRefGoogle Scholar
  36. Tilman D, Kilham SS, Kilham P (1982) Phytoplankton community ecology: the role of limiting nutrients. Ann Rev Ecol Syst 13: 349–372CrossRefGoogle Scholar
  37. Trüper HG, Schlegel HG (1964) Sulphur metabolism in Thiorhodaceae. I. Quantitative measurements on growing cells of Chromatium okenii. Antonie van Leeuwenhoek 30: 225–238CrossRefGoogle Scholar
  38. Visscher PT, Gemerden H van (1988) Growth of Chlorobium limicola f. thiosulfatophilum on polysulfides. In: Olsen JM, Ormerod JG, Amesz J, Stackebrandt E, Trüper HG (eds) Green photosynthetic bacteria. Plenum Press, New York, pp 287–294CrossRefGoogle Scholar
  39. Visscher PT, Nijburg JW, Gemerden H van (1990) Polysulfide utilization by Thiocapsa roseopersicina. Arch Microbiol 155: 75–81CrossRefGoogle Scholar
  40. Zaitseva GN, Gulikova OM, Kondrat'eva EN (1965) Biochemical changes in the cells of Chromatium minutissimum under photoautotrophic and photoheterotrophic conditions of growth. Microbiology 34: 499–504Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Jordi Mas
    • 1
  • Hans van Gemerden
    • 1
  1. 1.Department of MicrobiologyUniversity of GroningenHarenThe Netherlands

Personalised recommendations