1887

Abstract

NCIB 11883 was grown in glucose-limited continuous culture at low dilution rate. Whole cells transported glucose using an energy-dependent mechanism which exhibited an accumulation ratio > 2000. Three major periplasmic proteins were purified and their potential role as glucose-binding proteins (GBP) were investigated using equilibrium dialysis. Two of these, GBP1 ( 36500) and GBP2 ( 33500), bound -glucose with high affinity ( 0·23 and 0·07 μ respectively), whereas the third protein ( 30500) showed no binding ability. Competition experiments using various analogues showed that those which differed from glucose at C-6 (e.g. 6-chloro-6-deoxy--glucose and 6-deoxy--glucose) variably decreased the binding of glucose to both GBP1 and GBP2, whereas those which differed at C-4 (e.g. -galactose) were only effective with GBP1. The rate of glucose uptake and the concentration of the glucose-binding proteins increased in parallel during prolonged growth under glucose-limitation due to the emergence of new strains in which GBP1 (e.g. strain AR18) or GBP2 (e.g. strain AR9), but not both, was hyperproduced and accounted for at least 27% of the total cell protein. It is concluded that synthesizes two distinct periplasmic binding proteins which are involved in glucose transport, and that these proteins are maximally derepressed during growth under glucose limitation.

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1988-12-01
2021-10-23
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References

  1. Ames G.F.-L. 1986; Bacterial periplasmic transport systems: structure, mechanism, and evolution.. Annual Review of Biochemistry 55:397–425
    [Google Scholar]
  2. Beardsmore A.J., Aperghis P.N.G., Quayle J.R. 1982; Characterization of the assimilatory and dissimilatory pathways of carbon metabolism during growth of Methylophtius methylotrophus on methanol.. Journal of General Microbiology 128:1423–1439
    [Google Scholar]
  3. Brass J.M., Ehmann U., Bukau B. 1983; Reconstitution of maltose transport in Escherichia coli: conditions affecting import of maltose-binding protein into calcium-treated cells of maltose regulon mutants.. Journal of Bacteriology 155:97–106
    [Google Scholar]
  4. Brass M., Higgins C.F., Foley M., Rugman P.A., Birmingham J., Garland P.B. 1986; Lateral diffusion of proteins in the periplasm of Escherichia coli. . Journal of Bacteriology 165:787–794
    [Google Scholar]
  5. Collins S.H., Jarvis A.W., Lindsay R.J., Hamilton W.A. 1976; Proton movements coupled to lactate and alanine in Escherichia coli: isolation of mutants with altered stoichiometry in alanine transport.. Journal of Bacteriology 126:1232–1244
    [Google Scholar]
  6. Cornish A., Linton J.D., Jones C.W. 1987; The effect of growth conditions on the respiratory system of a succinoglucan-producing strain of Agrobacterium radiobacter. . Journal of General Microbiology 133:2971–2978
    [Google Scholar]
  7. Cornish A., Greenwood J.A., Jones C.W. 1988; The relationship between glucose transport and the production of succinoglucan exopolysaccharide by Agrobacterium radiobacter. . Journal of General Microbiology 134:3111–3122
    [Google Scholar]
  8. Durham D.R., Pmbbs P.V. Jr 1982; Fractionation and characterization of the phosphoenol- pyruvate: fructose 1 -phosphotransferase system from Pseudomonas aeruginosa. . Journal of Bacteriology 149:534–541
    [Google Scholar]
  9. Harder W., Dukhuizen L. 1983; Physiological responses to nutrient limitation.. Annual Review of Microbiology 37:1–23
    [Google Scholar]
  10. Harder W., Kuenen J.G., Matin A. 1977; Microbial selection in continuous culture.. Journal of Applied Bacteriology 43:1–24
    [Google Scholar]
  11. Henderson P.J.F. 1986; Active transport of sugars into Escherichia coli. . In Carbohydrate Metabolism in Cultured Cells pp 409–460 Morgan M. J. Edited by New York: Plenum Press;
    [Google Scholar]
  12. Henderson P.J.F., Macpherson A.J.S. 1986; Assay, genetics, proteins and reconstitution of proton-linked galactose, arabinose and xylose transport systems of Escherichia coli. . Methods in Enzymology 125:387–429
    [Google Scholar]
  13. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacterophage T4.. Nature, London 227:680–685
    [Google Scholar]
  14. Linton J.D., Woodard S., Gouldney D.G. 1986; The consequence of stimulating glucose dehydrogenase activity by the addition of PQQ on metabolite production by Agrobacterium radiobacter NCIB11883.. Applied Microbiology and Biotechnology 25:357–361
    [Google Scholar]
  15. Linton J.D., Evans M.W., Jones D.S., Gouldney D.G. 1987; Exocellular succinoglucan production by Agrobacterium radiobacter NCIB 11883.. Journal of General Microbiology 133:2961–2969
    [Google Scholar]
  16. Midgley M., Dawes E.A. 1973; The regulation of transport of glucose and methyl α-glucoside in Pseudomonas aeruginosa. . Biochemical Journal 132:141–154
    [Google Scholar]
  17. Neu H.C., Heppel L.A. 1965; The release of enzymes from Escherichia coli by osmotic shock and during the formation of sphaeroplasts.. Journal of Biological Chemistry 240:3685–3692
    [Google Scholar]
  18. Postma P.W. 1986; The bacterial phosphoenol- pyruvate: sugar phosphotransferase system of Escherichia coli and Salmonella typhimurium. . In Carbohydrate Metabolism in Cultured Cells pp 367–408 Morgan M. J. Edited by New York: Plenum Press;
    [Google Scholar]
  19. Quilter J.A., Jones C.W. 1984; The organisation of methanol dehydrogenase and c-type cytochromes on the respiratory membrane of Methylo- philus methylotrophus. . FEBS Letters 174:167–172
    [Google Scholar]
  20. Roberts B.K.., Midgley M., Dawes E.A. 1973; The metabolism of 2-oxogluconate by Pseudomonas aeruginosa. . Journal of General Microbiology 78:319–329
    [Google Scholar]
  21. Rutgers M., Teixera De Mattos M.J., Postma P.W., Van Dam K. 1987; Establishment of the steady state in glucose-limited chemostat cultures of Klebsiella pneumoniae. . Journal of General Microbiology 133:445–451
    [Google Scholar]
  22. Scatchard G. 1949; The attractions of proteins for small molecules and ions.. Annals of the New York Academy of Sciences 51:660–672
    [Google Scholar]
  23. Stinson M.W., Cohen M.A., Merrick J.M. 1977; Purification and properties of the periplasamic glucose-binding protein of Pseudomonas aeruginosa. . Journal of Bacteriology 131:672–681
    [Google Scholar]
  24. De Vries G.E., Van Brussel A.N., Qubpel A. 1982; Mechanism and regulation of glucose transport in Rhizobium leguminosarum. . Journal of Bacteriology 149:872–879
    [Google Scholar]
  25. Weber K., Osborne M. 1975; Proteins and sodium dodecyl sulphate: molecular weight determinations on polyacrylamide gels and related procedures.. In The Proteins, 3rd edn. pp 179–223 Neurath H., Hill R. L. Edited by London: Academic Press;
    [Google Scholar]
  26. Whiting P.H., Midgley M., Dawes E.A. 1976; The role of glucose limitation in the regulation of the transport of glucose, gluconate and 2-oxogluconate, and of glucose metabolism in Pseudomonas aeruginosa. . Journal of General Microbiology 92:304–310
    [Google Scholar]
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