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. 1983 Jun;154(3):1204–1214. doi: 10.1128/jb.154.3.1204-1214.1983

Regulation and characterization of the galactose-phosphoenolpyruvate-dependent phosphotransferase system in Lactobacillus casei.

B M Chassy, J Thompson
PMCID: PMC217592  PMID: 6406427

Abstract

Cells of Lactobacillus casei grown in media containing galactose or a metabolizable beta-galactoside (lactose, lactulose, or arabinosyl-beta-D-galactoside) were induced for a galactose-phosphoenolpyruvate-dependent phosphotransferase system (gal-PTS). This high-affinity system (Km for galactose, 11 microM) was inducible in eight strains examined, which were representative of all five subspecies of L. casei. The gal-PTS was also induced in strains defective in glucose- and lactose-phosphoenolpyruvate-dependent phosphotransferase systems during growth on galactose. Galactose 6-phosphate appeared to be the intracellular inducer of the gal-PTS. The gal-PTS was quite specific for D-galactose, and neither glucose, lactose, nor a variety of structural analogs of galactose caused significant inhibition of phosphotransferase system-mediated galactose transport in intact cells. The phosphoenolpyruvate-dependent phosphorylation of galactose in vitro required specific membrane and cytoplasmic components (including enzyme IIIgal), which were induced only by growth of the cells on galactose or beta-galactosides. Extracts prepared from such cells also contained an ATP-dependent galactokinase which converted galactose to galactose 1-phosphate. Our results demonstrate the separate identities of the gal-PTS and the lactose-phosphoenol-pyruvate-dependent phosphotransferase system in L. casei.

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Selected References

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  1. Bissett D. L., Anderson R. L. Lactose and D-galactose metabolism in group N streptococci: presence of enzymes for both the D-galactose 1-phosphate and D-tagatose 6-phosphate pathways. J Bacteriol. 1974 Jan;117(1):318–320. doi: 10.1128/jb.117.1.318-320.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bissett D. L., Anderson R. L. Lactose and D0galactose metabolism in Staphylococcus aureus: pathway of D-galactose 6-phosphate degradation. Biochem Biophys Res Commun. 1973 May 15;52(2):641–647. doi: 10.1016/0006-291x(73)90761-4. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Chassy B. M., Thompson J. Regulation of lactose-phosphoenolpyruvate-dependent phosphotransferase system and beta-D-phosphogalactoside galactohydrolase activities in Lactobacillus casei. J Bacteriol. 1983 Jun;154(3):1195–1203. doi: 10.1128/jb.154.3.1195-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crow V. L., Davey G. P., Pearce L. E., Thomas T. D. Plasmid linkage of the D-tagatose 6-phosphate pathway in Streptococcus lactis: effect on lactose and galactose metabolism. J Bacteriol. 1983 Jan;153(1):76–83. doi: 10.1128/jb.153.1.76-83.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Demko G. M., Blanton S. J., Benoit R. E. Heterofermentative carbohydrate metabolism of lactose-impaired mutants of Streptococcus lactis. J Bacteriol. 1972 Dec;112(3):1335–1345. doi: 10.1128/jb.112.3.1335-1345.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deutscher J., Beyreuther K., Sobek H. M., Stüber K., Hengstenberg W. Phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus: factor IIIlac, a trimeric phospho-carrier protein that also acts as a phase transfer catalyst. Biochemistry. 1982 Sep 28;21(20):4867–4873. doi: 10.1021/bi00263a006. [DOI] [PubMed] [Google Scholar]
  8. EFTHYMIOU C., HANSEN P. A. An antigenic analysis of Lactobacillus acidophilus. J Infect Dis. 1962 May-Jun;110:258–267. doi: 10.1093/infdis/110.3.258. [DOI] [PubMed] [Google Scholar]
  9. Hays J. B., Simoni R. D., Roseman S. Sugar transport. V. A trimeric lactose-specific phosphocarrier protein of the Staphylococcus aureus phosphotransferase system. J Biol Chem. 1973 Feb 10;248(3):941–956. [PubMed] [Google Scholar]
  10. Kashket E. R., Wilson T. H. Role of metabolic energy in the transport of -galactosides by Streptococcus lactis. J Bacteriol. 1972 Feb;109(2):784–789. doi: 10.1128/jb.109.2.784-789.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kornberg H. L., Reeves R. E. Inducible phosphoenolpyruvate-dependent hexose phosphotransferase activities in Escherichia coli. Biochem J. 1972 Aug;128(5):1339–1344. doi: 10.1042/bj1281339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LeBlanc D. J., Crow V. L., Lee L. N., Garon C. F. Influence of the lactose plasmid on the metabolism of galactose by Streptococcus lactis. J Bacteriol. 1979 Feb;137(2):878–884. doi: 10.1128/jb.137.2.878-884.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee L. J., Hansen J. B., Jagusztyn-Krynicka E. K., Chassy B. M. Cloning and expression of the beta-D-phosphogalactoside galactohydrolase gene of Lactobacillus casei in Escherichia coli K-12. J Bacteriol. 1982 Dec;152(3):1138–1146. doi: 10.1128/jb.152.3.1138-1146.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McKay L. L., Baldwin K. A. Stabilization of Lactose Metabolism in Streptococcus lactis C2. Appl Environ Microbiol. 1978 Aug;36(2):360–367. doi: 10.1128/aem.36.2.360-367.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morse M. L., Hill K. L., Egan J. B., Hengstenberg W. Metabolism of lactose by Staphylococcus aureus and its genetic basis. J Bacteriol. 1968 Jun;95(6):2270–2274. doi: 10.1128/jb.95.6.2270-2274.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Park Y. H., McKay L. L. Distinct galactose phosphoenolpyruvate-dependent phosphotransferase system in Streptococcus lactis. J Bacteriol. 1982 Feb;149(2):420–425. doi: 10.1128/jb.149.2.420-425.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Postma P. W., Roseman S. The bacterial phosphoenolpyruvate: sugar phosphotransferase system. Biochim Biophys Acta. 1976 Dec 14;457(3-4):213–257. doi: 10.1016/0304-4157(76)90001-0. [DOI] [PubMed] [Google Scholar]
  18. SHERMAN J. R. Rapid enzyme assay technique utilizing radioactive substrate, ion-exchange paper, and liquid scintillation counting. Anal Biochem. 1963 Jun;5:548–554. doi: 10.1016/0003-2697(63)90075-7. [DOI] [PubMed] [Google Scholar]
  19. Saier M. H., Jr Bacterial phosphoenolpyruvate: sugar phosphotransferase systems: structural, functional, and evolutionary interrelationships. Bacteriol Rev. 1977 Dec;41(4):856–871. doi: 10.1128/br.41.4.856-871.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schrecker Otto, Hengstenberg Wolfgang. Purification of the lactose specific factor III of the staphylococcal PEP dependent phosphotransferase system. FEBS Lett. 1971 Mar 16;13(4):209–212. doi: 10.1016/0014-5793(71)80537-9. [DOI] [PubMed] [Google Scholar]
  21. Schäfer A., Schrecker O., Hengstenberg W. The staphylococcal phosphoenolpyruvate-dependent phosphotransferase system. Purification and characterisation of the galactoside-specific membrane-component enzyme II. Eur J Biochem. 1981 Jan;113(2):289–294. doi: 10.1111/j.1432-1033.1981.tb05065.x. [DOI] [PubMed] [Google Scholar]
  22. Simoni R. D., Nakazawa T., Hays J. B., Roseman S. Sugar transport. IV. Isolation and characterization of the lactose phosphotransferase system in Staphylococcus aureus. J Biol Chem. 1973 Feb 10;248(3):932–940. [PubMed] [Google Scholar]
  23. Simoni R. D., Roseman S. Sugar transport. VII. Lactose transport in Staphylococcus aureus. J Biol Chem. 1973 Feb 10;248(3):966–974. [PubMed] [Google Scholar]
  24. Simoni R. D., Smith M. F., Roseman S. Resolution of a staphylococcal phosphotransferase system into four protein components and its relation to sugar transport. Biochem Biophys Res Commun. 1968 Jun 10;31(5):804–811. doi: 10.1016/0006-291x(68)90634-7. [DOI] [PubMed] [Google Scholar]
  25. Thomas T. D., Turner K. W., Crow V. L. Galactose fermentation by Streptococcus lactis and Streptococcus cremoris: pathways, products, and regulation. J Bacteriol. 1980 Nov;144(2):672–682. doi: 10.1128/jb.144.2.672-682.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thompson J., Chassy B. M. Novel phosphoenolpyruvate-dependent futile cycle in Streptococcus lactis: 2-deoxy-D-glucose uncouples energy production from growth. J Bacteriol. 1982 Sep;151(3):1454–1465. doi: 10.1128/jb.151.3.1454-1465.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thompson J., Chassy B. M. Uptake and metabolism of sucrose by Streptococcus lactis. J Bacteriol. 1981 Aug;147(2):543–551. doi: 10.1128/jb.147.2.543-551.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thompson J. Galactose transport systems in Streptococcus lactis. J Bacteriol. 1980 Nov;144(2):683–691. doi: 10.1128/jb.144.2.683-691.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thompson J. Lactose metabolism in Streptococcus lactis: phosphorylation of galactose and glucose moieties in vivo. J Bacteriol. 1979 Dec;140(3):774–785. doi: 10.1128/jb.140.3.774-785.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thompson J., Thomas T. D. Phosphoenolpyruvate and 2-phosphoglycerate: endogenous energy source(s) for sugar accumulation by starved cells of Streptococcus lactis. J Bacteriol. 1977 May;130(2):583–595. doi: 10.1128/jb.130.2.583-595.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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