1887

Abstract

Summary: Washed cells of prepared from a fructose-limited continuous culture (D 0.045 h) transported D-[UC]fructose in a linear manner for up to 4 min at a rate several-fold higher than the rate of fructose utilization by the growing culture. D-[UC]Fructose transport exhibited a high affinity for fructose ( < 1 μM) and was inhibited to varying extents by osmotic shock, by the uncoupling agent carbonyl cyanide , and by unlabelled sugars (D-fructose/D-mannose > D-ribose > D-sorbose > D-glucose/D-galactose/D-xylose; no inhibition by D-arabinose). Prolonged growth of in fructose-limited continuous culture led to the selection of a novel strain (AR100) which overproduced a fructose-binding protein (FBP) and showed an increased rate of fructose transport. FBP was purified from osmotic-shock fluid using anion-exchange fast protein liquid chromatography (FPLC). The monomeric protein (M 34200 by SDS-PAGE and 37700 by gel-filtration FPLC) bound D-[UC]-fructose stoichiometrically (1.17 nmol nmol FBP) and with high affinity (K 0.49 μM) as shown by equilibrium dialysis. Binding of D-[UC]fructose by FBP was variably inhibited by unlabelled sugars (D-fructose/D-mannose > D-ribose > D-sorbose; no inhibition by D-glucose, D-galactose or D-arabinose). The N-terminal amino acid sequence of FBP (ADTSVCLI-) was similar to that of several sugar-binding proteins from other species of bacteria. Fructose transport and FBP were variably induced in batch cultures of by growth on different carbon sources (D-fructose > D-ribose/D-mannose > D-glucose; no induction by succinate). An immunologically similar protein to FBP was produced by and various species of following growth on fructose. It is concluded that fructose is transported into and related organisms via a periplasmic fructose/mannose-binding-protein-dependent active-transport system, in contrast to the phosphotransferase system used by many other species of bacteria.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-10-2601
1995-10-01
2021-07-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/10/mic-141-10-2601.html?itemId=/content/journal/micro/10.1099/13500872-141-10-2601&mimeType=html&fmt=ahah

References

  1. Allenza P., Lee Y.N., Lessie T.G. 1982; Enzymes related to fructose utilisation in Pseudomonas cepacia. . J Bacteriol 150:1348–1356
    [Google Scholar]
  2. Argos P., Mahoney W.C., Hermodson M.A., Hanie M. 1981; Structural prediction of sugar-binding proteins functional in chemotaxis and transport.. J Biol Chem 256:4357–4361
    [Google Scholar]
  3. Arias A., Gardiol A., Martinez-Drets G. 1982; Transport and catabolism of D-mannose in Rhizobium meliloti. . J Bacteriol 151:1068–1072
    [Google Scholar]
  4. Brass M., Higgins C.F., Foley M., Rugman P.A., Birmingham i., Garland P.B. 1986; Lateral diffusion of proteins in the periplasm of Escherichia coli. . J Bacteriol 165:787–794
    [Google Scholar]
  5. Cornish A., Greenwood J.A., Jones C.W. 1988a; Binding- protein-dependent glucose transport by Agrobacterium radiobacter grown in continuous culture.. J Gen Microbiol 134:3099–3110
    [Google Scholar]
  6. Cornish A., Greenwood J.A., Jones C.W. 1988b; The relationship between glucose transport and the production of succinoglucan exopolysaccharide by Agrobacterium radiobacter. . J Gen Microbiol 134:3111–3122
    [Google Scholar]
  7. Cornish A., Greenwood J.A., Jones C.W. 1989; Binding- protein-dependent sugar transport by Agrobacterium radiobacter and A. tumefaciens grown in continuous culture.. J Gen Microbiol 135:3001–3013
    [Google Scholar]
  8. de Crecy-Lagard V., Lejeune P., Bouvet O.M.M., Danchin A. 1991; Identification of two fructose transport and phosphorylation pathways in Xanthomonas campestris pv. campestris.. Mol & Gen Genet 227:465–472
    [Google Scholar]
  9. Dills S.S., Apperson A., Schmidt M.R., Saier M.H. 1980; Carbohydrate transport in bacteria.. Microbiol Rev 44:385–418
    [Google Scholar]
  10. Dykhuizen D.E., Dean A.M., Hartl D.L. 1987; Metabolic flux and fitness.. Genetics 115:25–31
    [Google Scholar]
  11. Gardiol A., Arias A., Cervenansky C, Martinez-Drets G. 1980; Biochemical characterisation of a fructokinase mutant of Rhizobium meliloti. . J Bacteriol 144:12–16
    [Google Scholar]
  12. Glenn A.R., Arwas R., McKay I.A., Dilworth M.J. 1984; Fructose metabolism in wild-type, fructokinase-negative and re- vertant strains of Rhizobium leguminosarum. . J Gen Microbiol 130:213–237
    [Google Scholar]
  13. Greenwood J.A., Cornish A., Jones C.W. 1990; Binding protein-dependent lactose transport in Agrobacterium radiobacter. . J Bacteriol 172:1703–1710
    [Google Scholar]
  14. Guezzar M.E., Hornez J.-P., Courtois B., Derieux J.-C. 1988; Study of a fructose-negative mutant of Rhizobium meliloti. . FEMS Microbiol Lett 49:429–434
    [Google Scholar]
  15. Kornberg H.L. 1990; Fructose transport by Escherichia coli. . Philos Trans R Soc Fond B 326:505–513
    [Google Scholar]
  16. Lendenmann U., Egli T. 1995; Is Escherichia coli growing in glucose-limited chemostat culture able to utilize other sugars without lag?. Microbiology 141:71–78
    [Google Scholar]
  17. McLaughlin R.E., Hughes T.A. 1989; Transposon mutagenesis and complementation of the fructokinase gene in Rhizobium leguminosarum biovar trifolii. . J Gen Microbiol 135:2329–2334
    [Google Scholar]
  18. Postma P.W., Lengeler J. W., Jacobson G. R. . 1993; Phosphoenolpyruvate : carbohydrate phosphotransferase systems of bacteria.. Microbiol Rep 57:543–594
    [Google Scholar]
  19. Ronson C.W., Astwood P.M., Downie J.A. 1984; Molecular cloning and genetic organisation of C4-dicarboxylate transport genes from Rhizobium leguminosarum. . J Bacteriol 160:903–909
    [Google Scholar]
  20. Scatchard G. 1949; The attraction of proteins for small molecules and ions.. Ann N Y Acad Sci 51:660–672
    [Google Scholar]
  21. Spaink H.P., Aarts A., Stacey G., Bloemberg G.V., Lugtenberg B.J.J., Kennedy E.P. 1992; Detection and separation of Rhi^obium and Bradyrhizobium nod metabolites using thin-layer chromatography.. Mol Plant-Microbe Interact 5:72–80
    [Google Scholar]
  22. Williams S.G., Greenwood J.A., Jones C.W. 1990; Isolation of novel strains of Agrobacterium radiobacter with altered capacities for lactose metabolism and succinoglucan production.. J Gen Microbiol 136:2179–2188
    [Google Scholar]
  23. Williams S.G., Greenwood J.A., Jones C.W. 1992; Molecular analysis of the lac operon encoding the binding protein-dependent lactose transport system and β-galactosidase in Agrobacterium radiobacter. . Mol Microbiol 6:1755–1768
    [Google Scholar]
  24. Wylie J.L., Worobec E.A. 1993; substrate-specificity of the high-affinity glucose transport system of Pseudomonas aeruginosa. . Can J Microbiol 39:722–725
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-10-2601
Loading
/content/journal/micro/10.1099/13500872-141-10-2601
Loading

Data & Media loading...

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