Archives of Microbiology

, Volume 155, Issue 2, pp 153–158 | Cite as

Formation and role of glycine betaine in the moderate halophile Vibrio costicola

  • C. G. Choquet
  • I. Ahonkhai
  • M. Klein
  • D. J. Kushner
Original Papers


The moderate halophile Vibrio costicola, growing on a chemically-defined medium, transformed choline into glycine betaine (betaine) by the membrane-bound enzyme choline dehydrogenase and the cytoplasmic enzyme betainal (betaine aldehyde) dehydrogenase. Choline dehydrogenase was strongly induced and betainal dehydrogenase less strongly induced by choline. The formation of these enzymes was also regulated by the NaCl concentration of the growth medium, increasing with increasing NaCl concentrations. Intracellular betaine concentrations also increased with increasing choline and NaCl concentrations in the medium. This increase was almost completely blocked by chloramphenicol, which does not block the increase in salt-tolerant active transport on transfer from a low to a high salt concentration.

Choline dehydrogenase was inhibited by chloride salts of Na+, K+, and NH inf4 su+ , the inhibition being due to the Cl- ions. Betainal dehydrogenase was stimulated by 0.5 M salts and could function in up to 2.0 M salts.

Cells grew as well in the presence as in the absence of choline in 0.5 M and 1.0 M NaCl, but formed no intracellular betaine. Choline stimulated growth in 2.0 M NaCl and was essential for growth in 3.0 M NaCl. Thus, while betaine is important for some of the adaptations to high salt concentration by V. costicola, it by no means accounts for all of them.

Key words

Halophilic Vibrio costicola Choline dehydrogenase Betaine aldehyde dehydrogenase Betainal dehydrogenase Glycine betaine Transport 



chemically-defined minimal medium


proteose-peptone tryptone medium


sodium dodecyl sulfate


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bengis-GarberC, Gromet-ElhananZ (1979) Purification of the energy transducing adenosine triphosphatase complex from Rhodospirillum rubrum. Biochemistry 18:3577–3581CrossRefGoogle Scholar
  2. Bengis-GarberC, KushnerDJ (1981) Purification and properties of 5′-nucleotidase from the membrane of Vibrio costicola, a moderately halophilic bacterium. J Bacteriol 146:24–32PubMedPubMedCentralGoogle Scholar
  3. BorowitzkaLJ, BrownAD (1974) The salt relations of marine and halophilic species of the unicellular green alga, Dunaliella. The role of glycerol as a compatible solute. Arch Microbiol 96:37–52CrossRefGoogle Scholar
  4. BouillardL, LeRudulierD (1983) Nitrogen fixation under osmotic stress: enhancement of nitrogenase biosynthesis in Klebsiella pneumoniae by glycine betaine. Physiol Veg 21:447–457Google Scholar
  5. BrownAD (1976) Microbial water stress. Bacteriol Rev 40:803–846PubMedPubMedCentralGoogle Scholar
  6. ChoquetCG, KamekuraM, KushnerDJ (1989) In vitro protein synthesis by the moderate halophile Vibrio costicola: site of action of Cl- ions. J Bacteriol 171:880–886CrossRefGoogle Scholar
  7. ChoquetCG, KushnerDJ (1990) Use of natural mRNAs in the cell-free protein-synthesising systems of the moderate halophile Vibrio costicola. J Bacteriol 172:3462–3468CrossRefGoogle Scholar
  8. FlanneryWL, DoetschRN, HansenPA (1952) Salt desideratum of Vibrio costicolus, an obligate halophilic bacterium. I. Ionic replacement of sodium chloride requirement. J Bacteriol 64:713–717PubMedPubMedCentralGoogle Scholar
  9. ForsythMP, KushnerDJ (1970) Nutrition and distribution of salt response in populations of moderately halophilic bacteria. Can J Microbiol 16:253–261CrossRefGoogle Scholar
  10. GalinskyEA, PfeifferHP, TrüperHG (1985) 1,4,5,5-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid. A novel cyclic amino acid from halophilic phototrophic bacteria of the genus Ectothiorhodospira. Eur J Biochem 149:135–140CrossRefGoogle Scholar
  11. IkutaS, MatuuraK, ImamuraS, MisakiH, HoriutiY (1977) Oxidative pathway of choline to betaine in the soluble fraction prepared from Arthrobacter globiformis. J Biochem 82:157–163CrossRefGoogle Scholar
  12. ImhoffJF (1986) Osmoregulation and compatible solutes in eubacteria. FEMS Microbiol Rev 39:57–66CrossRefGoogle Scholar
  13. ImhoffJF, Rodriguez-ValeraF (1984) Betaine is the main compatible solute of halophilic eubacteria. J Bacteriol 160:478–479PubMedPubMedCentralGoogle Scholar
  14. KamekuraM, KushnerDJ (1984) Effect of chloride and glutamate ions on in vitro protein synthesis by the moderate halophile, Vibrio costicola. J Bacteriol 160:385–390PubMedPubMedCentralGoogle Scholar
  15. KamekuraM, WallaceR, HipkissAR, KushnerDJ (1985) Growth of Vibrio costicola and other moderate halophiles in a chemically defined minimal medium. Can J Microbiol 31:870–872CrossRefGoogle Scholar
  16. KushnerDJ (1978) Life in high salt and solute concentrations: halophilic bacteria. In: KushnerDJ (ed) Microbial life in extreme environments. Academic Press, London, pp 317–368Google Scholar
  17. KushnerDJ (1988) What is the “true” internal environment of halophilic and other bacteria? Can J Microbiol 34:482–486CrossRefGoogle Scholar
  18. KushnerDJ, KamekuraM (1988) Physiology of halophilic eubacteria. In: Rodriguez-ValeraF (ed) Halophilic bacteria. CRC Press, Boca Raton, FL, USA, pp 109–140Google Scholar
  19. KushnerDJ, HamaideF, MacLeodRA (1983) Development of saltresistant active transport in a moderately halophilic bacterium. J Bacteriol 153:1163–1171PubMedPubMedCentralGoogle Scholar
  20. LandfaldB, StromAR (1986) Choline-glycine betaine pathway confers high level of osmotic tolerance in Escherichia coli. J Bacteriol 165:849–855CrossRefGoogle Scholar
  21. LeRudulierD, BernardT (1986) Salt tolerance in Rhizobium: a possible role for betaines. FEMS Microbiol Rev 39:67–72CrossRefGoogle Scholar
  22. LowryOH, RosebroughNJ, FarrAL, RandallRJ (1951) Protein measurement with the Folin phenol reagent. J Mol Biol 193:265–275Google Scholar
  23. MeuryJ (1988) Glycine betaine reverses the effects of osmotic stress on DNA replication and cellular division in Escherichia coli. Arch Microbiol 149:232–239CrossRefGoogle Scholar
  24. NagasawaT, KawabataY, TaniY, OgataK (1975) Choline dehydrogenase of Pseudomonas aeruginosa A-16. Agric Biol Chem 7:1513–1514Google Scholar
  25. NagasawaT, KawabataY, TaniY, OgataK (1976) Purification and characterization of betaine aldehyde dehydrogenase from Pseudomonas aeruginosa A-16. Agric Biol Chem 9:1743–1749Google Scholar
  26. Ohte-FukuyamaM, MiyakeY, EmiS, YamanoT (1980) Identification and properties of the prosthetic group of choline oxidase from Alcaligenes sp. J Biochem 88:197–203Google Scholar
  27. PavlicekKA, YoppJH (1982) Betaine as a compatible solute in the complete relief of salt inhibition of glucose-6-phosphate dehydrogenase from a halophilic blue-green alga. Plant Physiol (Suppl) 69:58CrossRefGoogle Scholar
  28. PollardA, Wyn JonesRG (1979) Enzyme activities in concentrated solutions of glycinebetaine and other solutes. Plant 144:291–298CrossRefGoogle Scholar
  29. Rafaeli-EshkolD, Avi-DorY (1968) Studies on halotolerance in a moderately halophilic bacterium. Effect of betaine on salt resistance of the respiratory system. Biochem J 109:687–691CrossRefGoogle Scholar
  30. ShindlerDB, WydroRM, KushnerDJ (1977) Cell-bound cations of the moderately halophilic bacterium Vibrio costicola. J Bacteriol 130:698–703PubMedPubMedCentralGoogle Scholar
  31. SibleyMH, YoppJH (1987) Regulation of S-adenosylhomocysteine hydrolase in the halophilic cyanobacterium Aphanothece halophytica: a possible role in glycinebetaine biosynthesis. Arch Microbiol 149:43–46CrossRefGoogle Scholar
  32. SmithLT, PocardJA, BernardT, LeRudulierD (1988) Osmotic control of glycine betaine biosynthesis and degradation in Rhizobium meliloti. J Bacteriol 170:3142–3149CrossRefGoogle Scholar
  33. Trüper HG (1990) Halophily, taxonomy, phylogeny and nomenclature. Workshop on General and Applied Aspects of Halophilic Microorganisms, Alicante, Spain; In proceedings of this workshop (in press)Google Scholar
  34. WallJS, ChristiansonRJ, DimlerRJ, SentiFR (1960) Spectrophotometric determination of betaines and other quaternary nitrogen compounds as their periodides. Anal Chem 32:870–874CrossRefGoogle Scholar
  35. WarrSRC, ReedRH, StewartWDP (1984) Osmotic adjustment of cyanobacteria: the effects of NaCl, KCl, sucrose and glycine betaine on glutamine synthetase activity in a marine and a halotolerant strain. J Gen Microbiol 130:2169–2175Google Scholar
  36. WohlfarthA, SeverinJ, GalinskiEA (1990) The spectrum of compatible solutes in heterotrophic halophilic eubacteria of the family Halomonadaceae. J Gen Microbiol 136:705–712CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • C. G. Choquet
    • 1
  • I. Ahonkhai
    • 1
  • M. Klein
    • 1
  • D. J. Kushner
    • 1
  1. 1.Department of BiologyUniversity of OttawaOttawaCanada

Personalised recommendations