1887

Abstract

The dynamic response of cellular carbohydrates to a NaCl shock in biovar TA-1 (0·25 -NaCl) and SU-47 (0·4 -NaCl) grown in NaCl-free medium was investigated in non-growing cell cultures and in cell suspensions, using NMR. After transferring cells grown in a NaCl-free medium to a glutamic-acid-free medium containing mannitol and NaCl, both strains immediately responded to the increased osmotic pressure by augmenting the cellular trehalose content of the cell. Without mannitol in the medium trehalose synthesis was slower, but clearly detectable. Its synthesis paralleled the breakdown of the reserve materials glycogen and poly--hydroxybutyric acid (PHB). NMR experiments with 25-fold-concentrated cell suspensions using C-mannitol as substrate revealed that 15–20% of the trehalose synthesized was derived from mannitol, but 80–85% was from other sources. Trehalose was mainly formed from the internal pool of glycogen and/or PHB, whether mannitol was present or not, and reached 135 and 280 μg (mg cell protein) in the strains TA-1 and SU-47, respectively. At low osmolarity, intracellular trehalose was metabolized by strains TA-1 and SU-47. Intracellularly accumulated phosphoglycerol-substituted and neutral cyclic (1,2)--glucans of SU-47 cells grown in the absence of NaCl were neither degraded nor excreted after exposure to NaCl. Strain TA-1, which only makes neutral cyclic (1,2)--glucans, continued to synthesize and excrete cyclic (1,2)--glucans after exposure to NaCl. By using P-NMR, a sharp peak at 1·34 p.p.m. was present in cell suspensions of strain SU-47. This peak, representing glycerol-1-phosphate-substituted cyclic glucans, was absent in strain TA-1.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-12-3157
1993-12-01
2021-10-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/12/mic-139-12-3157.html?itemId=/content/journal/micro/10.1099/00221287-139-12-3157&mimeType=html&fmt=ahah

References

  1. Batley M., Redmond J.W., Djordjevic S.P., Rolfe B.G. 1987; Characterization of glycerophosphorylated cyclic β-(1,2)-glucans from a fast growing Rhizobium species. Biochimica et Biophysica Acta 901:119–126
    [Google Scholar]
  2. Botsford J.L., Lewis T.A. 1990; Osmoregulation in Rhizobium meliloti: production of glutamic acid in response to osmotic stress. Appied and Environmental Microbiology 56:488–494
    [Google Scholar]
  3. Braunegg G., Sonnleiter B., Lafferty R.M. 1978; A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass,. European Journal of Appied Microbiology 6:29–37
    [Google Scholar]
  4. Breedveld M.W., Zevenhuizen L.P.T.M., Zehnder A.J.B. 1990a; Excessive excretion of cyclic β-(1,2)-glucan by Rhizobium trifolii TA-1. Appied and Environmental Microbiology 56:2080–2086
    [Google Scholar]
  5. Breedveld M.W., Zevenhuizen L.P.T.M., Zehnder A.J.B. 1990b; Osmotically-induced oligo- and polysaccharide synthesis by Rhizobium meliloti SU-47. Journal of General Microbiology 136:2511–2519
    [Google Scholar]
  6. Breedveld M.W., Zevenhuizen L.P.T.M., Zehnder A.J.B. 1991; Osmotically-regulated trehalose accumulation and cyclic β-(1,2)-glucan excretion by Rhizobium leguminosarum biovar trifolii TA-1. Archives of Microbiology 156:501–506
    [Google Scholar]
  7. Burton R.M. 1957; The determination of glycerol and dihydroxy-acetone. Methods in Enzymology 3:246–249
    [Google Scholar]
  8. Claassen P.A.M., Dijkema C., Visser J., Zehnder A.J.B. 1986; In vivo13C-NMR analysis of acetate metabolism in Thiobacillus versutus under denitrifying conditions. Archives of Microbiology 146:227–232
    [Google Scholar]
  9. Csonka L.N. 1989; Physiological and genetic responses of bacteria to osmotic stress. Microbiological Reviews 53:121–147
    [Google Scholar]
  10. Dylan T., Helinski D.R., Ditta G.S. 1990; Hypoosmotic adaptation in Rhizobium meliloti requires β-(1,2)-glucan. Journal of Bacteriology 172:1400–1408
    [Google Scholar]
  11. Gloux K., Le Rudulier D. 1989; Transport and catabolism of proline betaine in salt-stressed Rhizobium meliloti. Archives of Microbiology 151:143–148
    [Google Scholar]
  12. Harris P.J., Henry R.J., Blakeney A.B., Stone B.A. 1984; An improved procedure for the methylation analyis of oligosaccharides and polysaccharides. Carbohydrate Research 127:59–73
    [Google Scholar]
  13. Hoelzle I., Streeter J.G. 1990; Increased accumulation of trehalose in rhizobia cultured under 1-percent oxygen. Appied and Environmental Microbiology 56:3213–3215
    [Google Scholar]
  14. Kennedy E.P., Rumley M.K. 1988; Osmotic regulation of biosynthesis of membrane-derived oligosaccharides in Escherichia coli. Journal of Bacteriology 170:2457–2461
    [Google Scholar]
  15. Le Rudulier D., Bernard T. 1986; Salt tolerance in rhizobia: a possible role for betaines. FEMS Microbiological Reviews 39:67–72
    [Google Scholar]
  16. Meikle A.J., Chudek J.A., Reed R.H., Gadd G.M. 1991; Natural abundance 13C-nuclear magnetic resonance spectroscopic analysis of acyclic polyol and trehalose accumulation by several yeast species in response to salt-stress. FEMS Microbiological Letters 82:163–168
    [Google Scholar]
  17. Miller K.J., Kennedy E.P., Reinhold V.N. 1986; Osmotic adaptation by Gram-negative bacteria: possible role for periplasmic oligosaccharides. Science 231:48–51
    [Google Scholar]
  18. Miller K.J., Gore R.S., Benesi A.J. 1988; Phosphoglycerol substituents present on the cyclic β-l,2-glucans of Rhizobium meliloti 1021 are derived from phosphatidylglycerol. Journal of Bacteriology 170:4569–4575
    [Google Scholar]
  19. Streeter J.G. 1985; Accumulation of α-α-trehalose by Rhizobium bacteria and bacteroids. Journal of Bacteriology 164:78–84
    [Google Scholar]
  20. Tombras Smith L., Smith G.M. 1989; An osmoregulated dipeptide in stressed Rhizobium meliloti. Journal of Bacteriology 172:4714–4717
    [Google Scholar]
  21. Tombras Smith L., Smith G.M., Madkour M.A. 1990; Osmoregulation in Agrobacterium tumefaciens: accumulation of a novel disaccharide is controlled by osmotic strength and glycine betaine. Journal of Bacteriology 172:6849–6855
    [Google Scholar]
  22. Trevelyan W.E., Harrison J.S. 1952; Studies on yeast metabolism. I. Fractionation and microdetermination of cell carbohydrates. Biochemical Journal 50:298–310
    [Google Scholar]
  23. Van Laere A. 1989; Trehalose, reserve and/or stress metabolite?. FEMS Microbiological Reviews 63:201–210
    [Google Scholar]
  24. Welsh D.T., Reed R.H., Herbert R.A. 1991; The role of trehalose in the osmoadaptation of Escherichia coli NCIB 9484. Interaction of trehalose, K+ and glutamate during osmoadaptation in continuous culture. Journal of General Microbiology 137:745–750
    [Google Scholar]
  25. Wiemken A. 1990; Trehalose in yeast, stress protectant rather than reserve carbohydrate. Antonie van Leeuwenhoek 58:209–217
    [Google Scholar]
  26. Zevenhuizen L.P.T.M. 1991; Levels of trehalose and glycogen in Arthrobacter globiformis under conditions of nutrient starvation and osmotic stress. Antonie van Leeuwenhoek 61:61–68
    [Google Scholar]
  27. Zevenhuizen L.P.T.M., Van Veldhuizen A., Fokkens R.H. 1990; Re-examination of cellular β-(1,2)-glucans of Rhizobiaceae: distribution of ring sizes and degrees of glycerol-1-phosphate substitution. Antonie van Leeuwenhoek 57:173–178
    [Google Scholar]
  28. Zorreguieta A., Cavaignac S., Geremia R.A., Ugalde R.A. 1990; Osmotic regulation of β-(1,2)-glucan synthesis in members of the family Rhizobiaceae. Journal of Bacteriology 172:4701–4704
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-12-3157
Loading
/content/journal/micro/10.1099/00221287-139-12-3157
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error