Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Oct;177(20):5806–5811. doi: 10.1128/jb.177.20.5806-5811.1995

Phosphorylating enzymes involved in glucose fermentation of Actinomyces naeslundii.

N Takahashi 1, S Kalfas 1, T Yamada 1
PMCID: PMC177402  PMID: 7592327

Abstract

Enzymatic activities involved in glucose fermentation of Actinomyces naeslundii were studied with glucose-grown cells from batch cultures. Glucose could be phosphorylated to glucose 6-phosphate by a glucokinase that utilized polyphosphate and GTP instead of ATP as a phosphoryl donor. Glucose 6-phosphate was further metabolized to the end products lactate, formate, acetate, and succinate through the Embden-Meyerhof-Parnas pathway. The phosphoryl donor for phosphofructokinase was only PPi. Phosphoglycerate kinase, pyruvate kinase, and acetate kinase coupled GDP as well as ADP, but P(i) compounds were not their phosphoryl acceptor. Cell extracts showed GDP-dependent activity of phosphoenolpyruvate carboxykinase, which assimilates bicarbonate and phosphoenolpyruvate into oxaloacetate, a precursor of succinate. Considerable amounts of GTP, polyphosphate, and PPi were found in glucose-fermenting cells, indicating that these compounds may serve as phosphoryl donors or acceptors in Actinomyces cells. PPi could be generated from UTP and glucose 1-phosphate through catalysis of UDP-glucose synthase, which provides UDP-glucose, a precursor of glycogen.

Full Text

The Full Text of this article is available as a PDF (213.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alves A. M., Euverink G. J., Hektor H. J., Hessels G. I., van der Vlag J., Vrijbloed J. W., Hondmann D., Visser J., Dijkhuizen L. Enzymes of glucose and methanol metabolism in the actinomycete Amycolatopsis methanolica. J Bacteriol. 1994 Nov;176(22):6827–6835. doi: 10.1128/jb.176.22.6827-6835.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bertagnolli B. L., Cook P. F. Kinetic mechanism of pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii. Biochemistry. 1984 Aug 28;23(18):4101–4108. doi: 10.1021/bi00313a014. [DOI] [PubMed] [Google Scholar]
  3. Bowden G. H., Ekstrand J., McNaughton B., Challacombe S. J. Association of selected bacteria with the lesions of root surface caries. Oral Microbiol Immunol. 1990 Dec;5(6):346–351. doi: 10.1111/j.1399-302x.1990.tb00439.x. [DOI] [PubMed] [Google Scholar]
  4. Brown A. T., Breeding L. C. Carbon dioxide metabolism by Actinomyces viscosus: pathways for succinate and aspartate production. Infect Immun. 1980 Apr;28(1):82–91. doi: 10.1128/iai.28.1.82-91.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buchanan B. B., Pine L. Path of glucose breakdown and cell yields of a facultative anaerobe, Actinomyces naeslundii. J Gen Microbiol. 1967 Feb;46(2):225–236. doi: 10.1099/00221287-46-2-225. [DOI] [PubMed] [Google Scholar]
  6. Clark J. E., Beegen H., Wood H. G. Isolation of intact chains of polyphosphate from "Propionibacterium shermanii" grown on glucose or lactate. J Bacteriol. 1986 Dec;168(3):1212–1219. doi: 10.1128/jb.168.3.1212-1219.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dawes E. A., Senior P. J. The role and regulation of energy reserve polymers in micro-organisms. Adv Microb Physiol. 1973;10:135–266. doi: 10.1016/s0065-2911(08)60088-0. [DOI] [PubMed] [Google Scholar]
  8. Duggleby R. G., Dennis D. T. Nicotinamide adenine dinucleotide-specific glyceraldehyde 3-phosphate dehydrogenase from Pisum sativum. Assay and steady state kinetics. J Biol Chem. 1974 Jan 10;249(1):167–174. [PubMed] [Google Scholar]
  9. Ellen R. P., Banting D. W., Fillery E. D. Longitudinal microbiological investigation of a hospitalized population of older adults with a high root surface caries risk. J Dent Res. 1985 Dec;64(12):1377–1381. doi: 10.1177/00220345850640121001. [DOI] [PubMed] [Google Scholar]
  10. Ellen R. P. Establishment and distribution of Actinomyces viscosus and Actinomyces naeslundii in the human oral cavity. Infect Immun. 1976 Nov;14(5):1119–1124. doi: 10.1128/iai.14.5.1119-1124.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ellen R. P., Segal D. N., Grove D. A. Relative proportions of Actinomyces viscosus and Actinomyces naeslundii in dental plaques collected from single sites. J Dent Res. 1978 Apr;57(4):550–550. doi: 10.1177/00220345780570040201. [DOI] [PubMed] [Google Scholar]
  12. Griffin J. B., Davidian N. M., Penniall R. Studies of phosphorus metabolism by isolated nuclei. VII. Identification of polyphosphate as a product. J Biol Chem. 1965 Nov;240(11):4427–4434. [PubMed] [Google Scholar]
  13. Hamilton I. R., Ellwood D. C. Carbohydrate metabolism by Actinomyces viscosus growing in continuous culture. Infect Immun. 1983 Oct;42(1):19–26. doi: 10.1128/iai.42.1.19-26.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heinonen J. K., Honkasalo S. H., Kukko E. I. A method for the concentration and for the colorimetric determination of nanomoles of inorganic pyrophosphate. Anal Biochem. 1981 Nov 1;117(2):293–300. doi: 10.1016/0003-2697(81)90725-9. [DOI] [PubMed] [Google Scholar]
  15. Janssen P. H., Morgan H. W. Glucose catabolism by Spirochaeta thermophila RI 19.B1. J Bacteriol. 1992 Apr;174(8):2449–2453. doi: 10.1128/jb.174.8.2449-2453.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johnson J. L., Moore L. V., Kaneko B., Moore W. E. Actinomyces georgiae sp. nov., Actinomyces gerencseriae sp. nov., designation of two genospecies of Actinomyces naeslundii, and inclusion of A. naeslundii serotypes II and III and Actinomyces viscosus serotype II in A. naeslundii genospecies 2. Int J Syst Bacteriol. 1990 Jul;40(3):273–286. doi: 10.1099/00207713-40-3-273. [DOI] [PubMed] [Google Scholar]
  17. Josse J. Constitutive inorganic pyrophosphatase of Escherichia coli. 1. Purification and catalytic properties. J Biol Chem. 1966 May 10;241(9):1938–1947. [PubMed] [Google Scholar]
  18. Kalfas S., Takahashi N., Yamada T. Initial catabolism of sorbitol in Actinomyces naeslundii and Actinomyces viscosus. Oral Microbiol Immunol. 1994 Dec;9(6):372–375. doi: 10.1111/j.1399-302x.1994.tb00288.x. [DOI] [PubMed] [Google Scholar]
  19. Komiyama K., Khandelwal R. L., Heinrich S. E. Glycogen synthetic and degradative activities by Actinomyces viscosus and Actinomyces naeslundii of root surface caries and noncaries sites. Caries Res. 1988;22(4):217–225. doi: 10.1159/000261109. [DOI] [PubMed] [Google Scholar]
  20. Kulaev I. S., Vagabov V. M. Polyphosphate metabolism in micro-organisms. Adv Microb Physiol. 1983;24:83–171. doi: 10.1016/s0065-2911(08)60385-9. [DOI] [PubMed] [Google Scholar]
  21. Loesche W. J., Syed S. A. Bacteriology of human experimental gingivitis: effect of plaque and gingivitis score. Infect Immun. 1978 Sep;21(3):830–839. doi: 10.1128/iai.21.3.830-839.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Macy J. M., Ljungdahl L. G., Gottschalk G. Pathway of succinate and propionate formation in Bacteroides fragilis. J Bacteriol. 1978 Apr;134(1):84–91. doi: 10.1128/jb.134.1.84-91.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Malcovati M., Kornberg H. L. Two types of pyruvate kinase in Escherichia coli K12. Biochim Biophys Acta. 1969 Apr 22;178(2):420–423. doi: 10.1016/0005-2744(69)90417-3. [DOI] [PubMed] [Google Scholar]
  24. Mertens E., Van Schaftingen E., Müller M. Presence of a fructose-2,6-bisphosphate-insensitive pyrophosphate: fructose-6-phosphate phosphotransferase in the anaerobic protozoa Tritrichomonas foetus, Trichomonas vaginalis and Isotricha prostoma. Mol Biochem Parasitol. 1989 Dec;37(2):183–190. doi: 10.1016/0166-6851(89)90150-3. [DOI] [PubMed] [Google Scholar]
  25. Moore W. E., Holdeman L. V., Smibert R. M., Cato E. P., Burmeister J. A., Palcanis K. G., Ranney R. R. Bacteriology of experimental gingivitis in children. Infect Immun. 1984 Oct;46(1):1–6. doi: 10.1128/iai.46.1.1-6.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moore W. E., Holdeman L. V., Smibert R. M., Good I. J., Burmeister J. A., Palcanis K. G., Ranney R. R. Bacteriology of experimental gingivitis in young adult humans. Infect Immun. 1982 Nov;38(2):651–667. doi: 10.1128/iai.38.2.651-667.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Brien W. E., Bowien S., Wood H. G. Isolation and characterization of a pyrophosphate-dependent phosphofructokinase from Propionibacterium shermanii. J Biol Chem. 1975 Nov 25;250(22):8690–8695. [PubMed] [Google Scholar]
  28. Phillips N. F., Horn P. J., Wood H. G. The polyphosphate- and ATP-dependent glucokinase from Propionibacterium shermanii: both activities are catalyzed by the same protein. Arch Biochem Biophys. 1993 Jan;300(1):309–319. doi: 10.1006/abbi.1993.1043. [DOI] [PubMed] [Google Scholar]
  29. Pollack J. D., Williams M. V. PPi-dependent phosphofructotransferase (phosphofructokinase) activity in the mollicutes (mycoplasma) Acholeplasma laidlawii. J Bacteriol. 1986 Jan;165(1):53–60. doi: 10.1128/jb.165.1.53-60.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Preiss J., Romeo T. Physiology, biochemistry and genetics of bacterial glycogen synthesis. Adv Microb Physiol. 1989;30:183–238. doi: 10.1016/s0065-2911(08)60113-7. [DOI] [PubMed] [Google Scholar]
  31. Reeves R. E., South D. J., Blytt H. J., Warren L. G. Pyrophosphate:D-fructose 6-phosphate 1-phosphotransferase. A new enzyme with the glycolytic function of 6-phosphofructokinase. J Biol Chem. 1974 Dec 25;249(24):7737–7741. [PubMed] [Google Scholar]
  32. Roberton A. M., Glucina P. G. Fructose 6-phosphate phosphorylation in Bacteroides species. J Bacteriol. 1982 Jun;150(3):1056–1060. doi: 10.1128/jb.150.3.1056-1060.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rogers A. H., de Jong M. H., Zilm P. S., van der Hoeven J. S. Estimation of growth parameters for some oral bacteria grown in continuous culture under glucose-limiting conditions. Infect Immun. 1986 Jun;52(3):897–901. doi: 10.1128/iai.52.3.897-901.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sabularse D. C., Anderson R. L. D-Fructose 2,6-bisphosphate: a naturally occurring activator for inorganic pyrophosphate:D-fructose-6-phosphate 1-phosphotransferase in plants. Biochem Biophys Res Commun. 1981 Dec 15;103(3):848–855. doi: 10.1016/0006-291x(81)90888-3. [DOI] [PubMed] [Google Scholar]
  35. Schedel M., Klemme J. H., Schlegel H. G. Regulation of C3-enzymes in facultative phototrophic bacteria: the cold-labile pyruvate kinase of Rhodopseudomonas sphaeroides. Arch Microbiol. 1975 May 5;103(3):237–245. doi: 10.1007/BF00436356. [DOI] [PubMed] [Google Scholar]
  36. Schobert P., Bowien B. Unusual C3 and C4 metabolism in the chemoautotroph Alcaligenes eutrophus. J Bacteriol. 1984 Jul;159(1):167–172. doi: 10.1128/jb.159.1.167-172.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Syed S. A., Loesche W. J., Pape H. L., Jr, grenier E. Predominant cultivable flora isolated from human root surface caries plaque. Infect Immun. 1975 Apr;11(4):727–731. doi: 10.1128/iai.11.4.727-731.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takahashi N., Iwami Y., Yamada T. Metabolism of intracellular polysaccharide in the cells of Streptococcus mutans under strictly anaerobic conditions. Oral Microbiol Immunol. 1991 Oct;6(5):299–304. doi: 10.1111/j.1399-302x.1991.tb00497.x. [DOI] [PubMed] [Google Scholar]
  39. Takahashi N., Kalfas S., Yamada T. The role of the succinate pathway in sorbitol fermentation by oral Actinomyces viscosus and Actinomyces naeslundii. Oral Microbiol Immunol. 1994 Aug;9(4):218–223. doi: 10.1111/j.1399-302x.1994.tb00061.x. [DOI] [PubMed] [Google Scholar]
  40. Takahashi N., Yamada T. Stimulatory effect of bicarbonate on the glycolysis of Actinomyces viscosus and its biochemical mechanism. Oral Microbiol Immunol. 1992 Jun;7(3):165–170. doi: 10.1111/j.1399-302x.1992.tb00530.x. [DOI] [PubMed] [Google Scholar]
  41. Tanzer J. M., Krichevsky M. I. Polyphosphate formation by caries-conducive streptococcus SL-I. Biochim Biophys Acta. 1970 Aug 14;215(2):368–376. doi: 10.1016/0304-4165(70)90036-x. [DOI] [PubMed] [Google Scholar]
  42. Tijssen J. P., Beekes H. W., Van Steveninck J. Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4',6-diamidino-2-phenylindole fluorescence. Biochim Biophys Acta. 1982 Dec 30;721(4):394–398. doi: 10.1016/0167-4889(82)90094-5. [DOI] [PubMed] [Google Scholar]
  43. Van Schaftingen E. Fructose 2,6-bisphosphate. Adv Enzymol Relat Areas Mol Biol. 1987;59:315–395. doi: 10.1002/9780470123058.ch7. [DOI] [PubMed] [Google Scholar]
  44. Wilke D., Schlegel H. G. Regulation of the pyruvate kinase from Alcaligenes eutrophus H 16 in vitro and in vivo. Arch Microbiol. 1975 Oct 27;105(2):109–115. doi: 10.1007/BF00447123. [DOI] [PubMed] [Google Scholar]
  45. Wood H. G., Goss N. H. Phosphorylation enzymes of the propionic acid bacteria and the roles of ATP inorganic pyrophosphate, and polyphosphates. Proc Natl Acad Sci U S A. 1985 Jan;82(2):312–315. doi: 10.1073/pnas.82.2.312. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES