Skip to main content
SpringerLink
Log in

Transport and catabolism of proline betaine in salt-stressed Rhizobium meliloti

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Exogenous proline betaine (N,N-dimethylproline or stachydrine) highly stimulated the growth rate of Rhizobium meliloti, in media of inhibitory concentration of NaCl whereas proline was ineffective. High levels of proline betaine uptake occurred in cells grown in media of elevated osmotic strength; on the contrary, only low activity was found in cells grown in minimal medium. The apparent K m was 10 μM with a maximal transport rate of 25 nmol min-1 mg-1 of protein in 0.3 M NaCl-grown cells. The concentrative transport was totally abolished by KCN (2 mM), 2,4-dinitrophenol (2 mM), and carbonyl cyanide-m-chlorophenyl hydrazone (CCCP 10 μM) but was insensitive to arsenate (5 mM). Glycine betaine was a very potent inhibitor of proline betaine uptake while proline was not. Proline betaine transport was not reduced in osmotically shocked cells and no proline betaine binding activity was detected in the crude periplasmic shock fluid. In the absence of salt stress, Rhizobium meliloti actively catabolized proline betaine but this catabolism was blocked by increasing the osmotic strength of the medium. The osmolarity in the growth medium regulates the use of proline betaine either as a carbon and nitrogen source or as an osmoprotectant.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

LAS:

lactate-aspartate-salts

MSY:

mannitol-salts-yeast

CCCP:

carbonyl cyanide-m-chlorophenyl hydrazone

DCCD:

dicyclohexylcarbodiimide

KCN:

potassium cyanide

Hepes:

4-(2-hydroxyethyl)-1-piperzine-ethanesulphonic acid

References

  • Ames GFL (1988) Structure and mechanism of bacterial periplasmic transport system. J Bioenerg Biomembr 20:1–18

    Google Scholar 

  • Argast M, Boos W (1979) Purification and properties of the sn-glycerol-3-phosphate-binding protein of Escherichia coli. J Biol Chem 254:10931–10935

    Google Scholar 

  • Barron A, Jung JU, Villarejo M (1987) Purification and characterization of a glycine betaine binding protein from Escherichia coli. J Biol Chem 262:11841–11846

    Google Scholar 

  • Bernard T, Pocard JA, Perroud B, Le Rudulier D (1986) Variations in the response of salt-stressed Rhizobium strains to betaines. Arch Microbiol 143:359–364

    Google Scholar 

  • Bostford JL (1984) Osmoregulation in Rhizobium meliloti: inhibition of growth by salts. Arch Microbiol 137:124–127

    Google Scholar 

  • Cairney J, Booth IR, Higgins CF (1985) Osmoregulation of gene expression in Salmonella typhimurium: proU encodes an osmotically induced betaine transport system. J Bacteriol 164:1224–1232

    Google Scholar 

  • Chambers ST, Kunin CM (1987) Isolation of glycine betaine and proline betaine from human urine. Assessment of their role as osmoprotective agents for bacteria and the kidney. J Clin Invest 79:731–737

    Google Scholar 

  • Corti UA (1949) Über Ergebnisse der Methylierung von Tyrosine mit Dimethylsulfate und einige Derivate des N-Methyltyramin. Helv Chim Acta 32:681–686

    Google Scholar 

  • Csonka LN (1981) Proline over-production results in enhanced osmotolerance in Salmonella typhimurium. Mol Gen Genet 182:82–86

    Google Scholar 

  • Csonka LN (1982) A third L-proline permease in Salmonella typhimurium which functions in media of elevated osmotic strength. J Bacteriol 151:1433–1443

    Google Scholar 

  • Dahl MK, Manson MD (1985) Interspecific reconstitution of maltose transport and chemotaxis in Escherichia coli with maltose-binding protein from various enteric bacteria. J Bacteriol 164:1051–1063

    Google Scholar 

  • De Maagd RA, Lugtenberg B (1986) Fractionation of Rhizobium leguminosarum cells into outer membrane, cytoplasmic membrane, periplasmic, and cytoplasmic components. J Bacteriol 167:1083–1085

    Google Scholar 

  • Glenn AR, Dilworth MJ (1979) An examination of Rhizobium leguminosarum for the production of extracellular and periplasmic proteins. J Gen Microbiol 112:405–409

    Google Scholar 

  • Gorham J (1984) Separation of plant betaines and their sulphur analogues by cation-exchange high-performance liquid chromatography. J Chromatogr 287:345–351

    Google Scholar 

  • Gowrishankar J (1985) Identification of osmoresponsive genes in Escherichia coli: evidence for participation of potassium and proline transport systems in osmoregulation. J Bacteriol 166:331–333

    Google Scholar 

  • Higgins CF, Cairney J, Stirling DA, Sutherland L, Booth IR (1987a) Osmotic regulation of gene expression: ionic strength as an intracellular signal? Trends Biochem Sci 12:339–344

    Google Scholar 

  • Higgins CF, Sutherland L, Cairney J, Booth IR (1987b) The osmotically regulated proU locus of Salmonella typhimurium encodes a periplasmic betaine-binding protein. J Gen Microbiol 133:305–310

    Google Scholar 

  • Hua SST, Tsai VY, Lichens GM, Noma AT (1982) Accumulation of amino acids in Rhizobium sp strain WR1001 in response to sodium chloride salinity. Appl Environ Microbiol 44:135–140

    Google Scholar 

  • Hutkins RW, Ellefson WL, Kashket ER (1987) Betaine transport imparts osmotolerance on a strain of Lactobacillus acidophilus. Appl Environ Microbiol 53:2275–2281

    Google Scholar 

  • Ikutani Y (1968) Studies of the N-oxides of N,N-dialkylamino acids. I. The synthesis of N,N-dimethyl neutral amino acids and corresponding N-oxides. Bull Chem Soc Jpn 41:1679–1681

    Google Scholar 

  • Le Rudulier D, Bouillard L (1983) Glycine betaine: an osmotic effector in Klebsiella pneumoniae. Appl Environ Microbiol 46:152–159

    Google Scholar 

  • Le Rudulier D, Yang SS, Csonka LN (1982) Nitrogen fixation in Klebsiella pneumoniae during osmotic stress. Effect of exogenous proline or a proline overproducing plasmid. Biochim Biophys Acta 719:273–283

    Google Scholar 

  • Le Rudulier D, Bernard T, Pocard JA, Goas G (1983) Accroissement de l'osmotolérance chez Rhizobium meliloti par la glycine bétaine et la proline bétaine. CR Acad Sc Paris 297:155–160

    Google Scholar 

  • Le Rudulier D, Strøm AR, Dandekar AM, Smith LT, Valentine RC (1984) Molecular biology of osmoregulation. Science 224: 1064–1068

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • May G, Faatz E, Villarejo M, Bremer E (1986) Binding protein dependent transport of glycine betaine and its osmotic regulation in Escherichia coli K 12. Mol Gen Genet 205:225–233

    Google Scholar 

  • Neu HC, Heppel LA (1965) The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 240:3685–3692

    Google Scholar 

  • Nobile S, Deshusses J (1986) Transport of γ-butyrobetaine in an Agrobacterium species isolated from soil. J Bacteriol 168:780–784

    Google Scholar 

  • O'Gara F, Shanmugan KT (1976) Regulation of nitrogen fixation by Rhizobia: export of fixed N2 as NH +4 . Biochim Biophys Acta 437:313–321

    Google Scholar 

  • Perroud B, Le Rudulier D (1985) Glycine betaine transport in Escherichia coli: osmotic modulation. J Bacteriol 161:393–401

    Google Scholar 

  • Pettigrew GW, Smith GM (1977) Novel N-terminal protein blocking group identified as dimethylproline. Nature (London) 265:661–662

    Google Scholar 

  • Pierce SK, Edwards SC, Mazzocchi PH, Klingler LJ, Warten MK (1984) Proline betaine: a unique osmolyte in an extremely euryhaline osmoconformer. Biol Bull 167:495–500

    Google Scholar 

  • Sethi LT, Carew DP (1974) Growth and betaine formation in Medicago sativa tissue culture. Phytochemistry 13:321–324

    Google Scholar 

  • Smith LT, Pocard JA, Bernard T, Le Rudulier D (1988) Osmotic control of glycine betaine biosynthesis and degradation in Rhizobium meliloti. J Bacteriol 170:3142–3149

    Google Scholar 

  • Stock JB, Rauch B, Roseman S (1977) Periplasmic space in Salmonella typhimurium and Escherichia coli. J Biol Chem 252:7850–7861

    Google Scholar 

  • Sutherland L, Cairney J, Elmore MJ, Booth IR, Higgins CF (1986) Osmotic regulation of transcription: induction of the proU betaine transport gene is dependent on accumulation of intracellular potassium. J Bacteriol 168:805–814

    Google Scholar 

  • Tepfer D, Goldmann A, Pamboukdjian N, Maille M, Lepingle A, Chevalier D, Dénarié J, Rosenberg C (1988) A plasmid of Rhizobium meliloti 41 encodes catabolism of two compounds from root exudate of Calystegium sepium. J Bacteriol 170:1153–1161

    Google Scholar 

  • Yap SF, Lim ST (1983) Response of Rhizobium sp UMKL 20 to chloride stress. Arch Microbiol 135:224–228

    Google Scholar 

  • Yelton MM, Yang SS, Edie SA, Lim ST (1983) Characterization of an effective salt tolerance fast growing strain of Rhizobium japonicum. J Gen Microbiol 129:1537–1547

    Google Scholar 

Download references

Author information

Author notes

  1. Daniel Le Rudulier

    Present address: UA CNRS 1114, Faculté des Sciences et des Techniques, Université de Nice, Parc Valrose, F-06034, Nice Cédex, France

Authors and Affiliations

  1. Laboratoire de Microbiologie et Physiologie des Symbioses, UA CNRS 256, Université de Rennes I, F-35042, Rennes Cédex, France

    Karine Gloux & Daniel Le Rudulier

Authors
  1. Karine Gloux
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel Le Rudulier
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gloux, K., Le Rudulier, D. Transport and catabolism of proline betaine in salt-stressed Rhizobium meliloti . Arch. Microbiol. 151, 143–148 (1989). https://doi.org/10.1007/BF00414429

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00414429

Key words

Search

Navigation