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. 1985 Apr;162(1):217–223. doi: 10.1128/jb.162.1.217-223.1985

Lactose metabolism in Streptococcus lactis: studies with a mutant lacking glucokinase and mannose-phosphotransferase activities.

J Thompson, B M Chassy, W Egan
PMCID: PMC218977  PMID: 3920203

Abstract

A mutant of Streptococcus lactis 133 has been isolated that lacks both glucokinase and phosphoenolpyruvate-dependent mannose-phosphotransferase (mannose-PTS) activities. The double mutant S. lactis 133 mannose-PTSd GK- is unable to utilize either exogenously supplied or intracellularly generated glucose for growth. Fluorographic analyses of metabolites formed during the metabolism of [14C]lactose labeled specifically in the glucose or galactosyl moiety established that the cells were unable to phosphorylate intracellular glucose. However, cells of S. lactis 133 mannose-PTSd GK- readily metabolized intracellular glucose 6-phosphate, and the growth rates and cell yield of the mutant and parental strains on sucrose were the same. During growth on lactose, S. lactis 133 mannose-PTSd GK- fermented only the galactose moiety of the disaccharide, and 1 mol of glucose was generated per mol of lactose consumed. For an equivalent concentration of lactose, the cell yield of the mutant was 50% that of the wild type. The specific rate of lactose utilization by growing cells of S. lactis 133 mannose-PTSd GK- was ca. 50% greater than that of the wild type, but the cell doubling times were 70 and 47 min, respectively. High-resolution 31P nuclear magnetic resonance studies of lactose transport by starved cells of S. lactis 133 and S. lactis 133 mannose-PTSd GK- showed that the latter cells contained elevated lactose-PTS activity. Throughout exponential growth on lactose, the mutant maintained an intracellular steady-state glucose concentration of 100 mM. We conclude from our data that phosphorylation of glucose by S. lactis 133 can be mediated by only two mechanisms: (i) via ATP-dependent glucokinase, and (ii) by the phosphoenolpyruvate-dependent mannose-PTS system.

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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. Chassy B. M., Porter E. V. Initial characterization of sucrose-6-phosphate hydrolase from Streptococcus mutans and its apparent identity with intracellular invertase. Biochem Biophys Res Commun. 1979 Jul 12;89(1):307–314. doi: 10.1016/0006-291x(79)90979-3. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Crow V. L., Thomas T. D. D-tagatose 1,6-diphosphate aldolase from lactic streptococci: purification, properties, and use in measuring intracellular tagatose 1,6-diphosphate. J Bacteriol. 1982 Aug;151(2):600–608. doi: 10.1128/jb.151.2.600-608.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curtis S. J., Epstein W. Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase. J Bacteriol. 1975 Jun;122(3):1189–1199. doi: 10.1128/jb.122.3.1189-1199.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FRAENKEL D. G., FALCOZ-KELLY F., HORECKER B. L. THE UTILIZATION OF GLUCOSE 6-PHOSPHATE BY GLUCOKINASELESS AND WILD-TYPE STRAINS OF ESCHERICHIA COLI. Proc Natl Acad Sci U S A. 1964 Nov;52:1207–1213. doi: 10.1073/pnas.52.5.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farrow J. A. Lactose hydrolysing enzymes in Streptococcus lactis and Streptococcus cremoris and also in some other species of streptococci. J Appl Bacteriol. 1980 Dec;49(3):493–503. doi: 10.1111/j.1365-2672.1980.tb04724.x. [DOI] [PubMed] [Google Scholar]
  8. Fordyce A. M., Crow V. L., Thomas T. D. Regulation of product formation during glucose or lactose limitation in nongrowing cells of Streptococcus lactis. Appl Environ Microbiol. 1984 Aug;48(2):332–337. doi: 10.1128/aem.48.2.332-337.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Harlander S. K., McKay L. L., Schachtele C. F. Molecular cloning of the lactose-metabolizing genes from Streptococcus lactis. Appl Environ Microbiol. 1984 Aug;48(2):347–351. doi: 10.1128/aem.48.2.347-351.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson K. G., McDonald I. J. Beta-D-phosphogalactoside galactohydrolase from Streptococcus cremoris HP: purification and enzyme properties. J Bacteriol. 1974 Feb;117(2):667–674. doi: 10.1128/jb.117.2.667-674.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mason P. W., Carbone D. P., Cushman R. A., Waggoner A. S. The importance of inorganic phosphate in regulation of energy metabolism of Streptococcus lactis. J Biol Chem. 1981 Feb 25;256(4):1861–1866. [PubMed] [Google Scholar]
  12. McKay L. L., Walter L. A., Sandine W. E., Elliker P. R. Involvement of phosphoenolpyruvate in lactose utilization by group N streptococci. J Bacteriol. 1969 Aug;99(2):603–610. doi: 10.1128/jb.99.2.603-610.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McKay L., Miller A., 3rd, Sandine W. E., Elliker P. R. Mechanisms of lactose utilization by lactic acid streptococci: enzymatic and genetic analyses. J Bacteriol. 1970 Jun;102(3):804–809. doi: 10.1128/jb.102.3.804-809.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Porter E. V., Chassy B. M., Holmlund C. E. Purification and kinetic characterization of a specific glucokinase from Streptococcus mutans OMZ70 cells. Biochim Biophys Acta. 1982 Dec 20;709(2):178–186. doi: 10.1016/0167-4838(82)90459-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Robillard G. T. The enzymology of the bacterial phosphoenolpyruvate-dependent sugar transport systems. Mol Cell Biochem. 1982 Jul 7;46(1):3–24. doi: 10.1007/BF00215577. [DOI] [PubMed] [Google Scholar]
  17. St Martin E. J., Wittenberger C. L. Regulation and function of sucrose 6-phosphate hydrolase in Streptococcus mutans. Infect Immun. 1979 Nov;26(2):487–491. doi: 10.1128/iai.26.2.487-491.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tanaka S., Fraenkel D. G., Lin E. C. The enzymatic lesion of strain MM-6, a pleiotropic carbohydrate-negative mutant of Escherichia coli. Biochem Biophys Res Commun. 1967 Apr 7;27(1):63–67. doi: 10.1016/s0006-291x(67)80040-8. [DOI] [PubMed] [Google Scholar]
  19. Thomas T. D. Regulation of lactose fermentation in group N streptococci. Appl Environ Microbiol. 1976 Oct;32(4):474–478. doi: 10.1128/aem.32.4.474-478.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Thompson J., Chassy B. M. Intracellular phosphorylation of glucose analogs via the phosphoenolpyruvate: mannose-phosphotransferase system in Streptococcus lactis. J Bacteriol. 1985 Apr;162(1):224–234. doi: 10.1128/jb.162.1.224-234.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Thompson J. In vivo regulation of glycolysis and characterization of sugar: phosphotransferase systems in Streptococcus lactis. J Bacteriol. 1978 Nov;136(2):465–476. doi: 10.1128/jb.136.2.465-476.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Thompson J., Saier M. H., Jr Regulation of methyl-beta-d-thiogalactopyranoside-6-phosphate accumulation in Streptococcus lactis by exclusion and expulsion mechanisms. J Bacteriol. 1981 Jun;146(3):885–894. doi: 10.1128/jb.146.3.885-894.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Thompson J., Torchia D. A. Use of 31P nuclear magnetic resonance spectroscopy and 14C fluorography in studies of glycolysis and regulation of pyruvate kinase in Streptococcus lactis. J Bacteriol. 1984 Jun;158(3):791–800. doi: 10.1128/jb.158.3.791-800.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. West I. C. Lactose transport coupled to proton movements in Escherichia coli. Biochem Biophys Res Commun. 1970 Nov 9;41(3):655–661. doi: 10.1016/0006-291x(70)90063-x. [DOI] [PubMed] [Google Scholar]

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