Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Nov;176(22):6827–6835. doi: 10.1128/jb.176.22.6827-6835.1994

Enzymes of glucose and methanol metabolism in the actinomycete Amycolatopsis methanolica.

A M Alves 1, G J Euverink 1, H J Hektor 1, G I Hessels 1, J van der Vlag 1, J W Vrijbloed 1, D Hondmann 1, J Visser 1, L Dijkhuizen 1
PMCID: PMC197050  PMID: 7961441

Abstract

The actinomycete Amycolatopsis methanolica was found to employ the normal bacterial set of glycolytic and pentose phosphate pathway enzymes, except for the presence of a PPi-dependent phosphofructokinase (PPi-PFK) and a 3-phosphoglycerate mutase that is stimulated by 2,3-bisphosphoglycerate. Screening of a number of actinomycetes revealed PPi-PFK activity only in members of the family Pseudonocardiaceae. The A. methanolica PPi-PFK and 3-phosphoglycerate mutase enzymes were purified to homogeneity. PPi-PFK appeared to be insensitive to the typical effectors of ATP-dependent PFK enzymes. Nevertheless, strong N-terminal amino acid sequence homology was found with ATP-PFK enzymes from other bacteria. The A. methanolica pyruvate kinase was purified over 250-fold and characterized as an allosteric enzyme, sensitive to inhibition by P(i) and ATP but stimulated by AMP. By using mutants, evidence was obtained for the presence of transketolase isoenzymes functioning in the pentose phosphate pathway and ribulose monophosphate cycle during growth on glucose and methanol, respectively.

Full text

PDF
6827

Selected References

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

  1. Baumann P., Baumann L. Catabolism of D-fructose and D-ribose by Pseudomonas doudoroffii. I. Physiological studies and mutant analysis. Arch Microbiol. 1975 Nov 7;105(3):225–240. doi: 10.1007/BF00447141. [DOI] [PubMed] [Google Scholar]
  2. 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Bystrykh L. V., Vonck J., van Bruggen E. F., van Beeumen J., Samyn B., Govorukhina N. I., Arfman N., Duine J. A., Dijkhuizen L. Electron microscopic analysis and structural characterization of novel NADP(H)-containing methanol: N,N'-dimethyl-4-nitrosoaniline oxidoreductases from the gram-positive methylotrophic bacteria Amycolatopsis methanolica and Mycobacterium gastri MB19. J Bacteriol. 1993 Mar;175(6):1814–1822. doi: 10.1128/jb.175.6.1814-1822.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlisle S. M., Blakeley S. D., Hemmingsen S. M., Trevanion S. J., Hiyoshi T., Kruger N. J., Dennis D. T. Pyrophosphate-dependent phosphofructokinase. Conservation of protein sequence between the alpha- and beta-subunits and with the ATP-dependent phosphofructokinase. J Biol Chem. 1990 Oct 25;265(30):18366–18371. [PubMed] [Google Scholar]
  5. Chou A. C., Wilson J. E. Hexikinase of rat brain. Methods Enzymol. 1975;42:20–25. doi: 10.1016/0076-6879(75)42086-9. [DOI] [PubMed] [Google Scholar]
  6. Euverink G. J., Hessels G. I., Vrijbloed J. W., Coggins J. R., Dijkhuizen L. Purification and characterization of a dual function 3-dehydroquinate dehydratase from Amycolatopsis methanolica. J Gen Microbiol. 1992 Nov;138(11):2449–2457. doi: 10.1099/00221287-138-11-2449. [DOI] [PubMed] [Google Scholar]
  7. Fothergill-Gilmore L. A., Michels P. A. Evolution of glycolysis. Prog Biophys Mol Biol. 1993;59(2):105–235. doi: 10.1016/0079-6107(93)90001-z. [DOI] [PubMed] [Google Scholar]
  8. French B. A., Chang S. H. Nucleotide sequence of the phosphofructokinase gene from Bacillus stearothermophilus and comparison with the homologous Escherichia coli gene. Gene. 1987;54(1):65–71. doi: 10.1016/0378-1119(87)90348-9. [DOI] [PubMed] [Google Scholar]
  9. Gracy R. W., Tilley B. E. Phosphoglucose isomerase of human erythrocytes and cardiac tissue. Methods Enzymol. 1975;41:392–400. doi: 10.1016/s0076-6879(75)41086-2. [DOI] [PubMed] [Google Scholar]
  10. Heijthuijsen J. H., Hansen T. A. Betaine fermentation and oxidation by marine desulfuromonas strains. Appl Environ Microbiol. 1989 Apr;55(4):965–969. doi: 10.1128/aem.55.4.965-969.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hellinga H. W., Evans P. R. Nucleotide sequence and high-level expression of the major Escherichia coli phosphofructokinase. Eur J Biochem. 1985 Jun 3;149(2):363–373. doi: 10.1111/j.1432-1033.1985.tb08934.x. [DOI] [PubMed] [Google Scholar]
  12. Hondmann D. H., Visser J. Screening method for large numbers of dye-adsorbents for enzyme purification. J Chromatogr. 1990 Jun 27;510:155–164. doi: 10.1016/s0021-9673(01)93749-5. [DOI] [PubMed] [Google Scholar]
  13. Kemp R. G., Tripathi R. L. Pyrophosphate-dependent phosphofructo-1-kinase complements fructose 1,6-bisphosphatase but not phosphofructokinase deficiency in Escherichia coli. J Bacteriol. 1993 Sep;175(17):5723–5724. doi: 10.1128/jb.175.17.5723-5724.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ladror U. S., Gollapudi L., Tripathi R. L., Latshaw S. P., Kemp R. G. Cloning, sequencing, and expression of pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii. J Biol Chem. 1991 Sep 5;266(25):16550–16555. [PubMed] [Google Scholar]
  15. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  16. Mertens E. ATP versus pyrophosphate: glycolysis revisited in parasitic protists. Parasitol Today. 1993 Apr;9(4):122–126. doi: 10.1016/0169-4758(93)90169-g. [DOI] [PubMed] [Google Scholar]
  17. Mertens E. Pyrophosphate-dependent phosphofructokinase, an anaerobic glycolytic enzyme? FEBS Lett. 1991 Jul 8;285(1):1–5. doi: 10.1016/0014-5793(91)80711-b. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Ozaki H., Shiio I. Regulation of the TCA and glyoxylate cycles in Brevibacterium flavum. II. Regulation of phosphoenolpyruvate carboxylase and pyruvate kinase. J Biochem. 1969 Sep;66(3):297–311. doi: 10.1093/oxfordjournals.jbchem.a129148. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Sakai H., Ohta T. Molecular cloning and nucleotide sequence of the gene for pyruvate kinase of Bacillus stearothermophilus and the production of the enzyme in Escherichia coli. Evidence that the genes for phosphofructokinase and pyruvate kinase constitute an operon. Eur J Biochem. 1993 Feb 1;211(3):851–859. doi: 10.1111/j.1432-1033.1993.tb17618.x. [DOI] [PubMed] [Google Scholar]
  22. Spring T. G., Wold F. Enolase from Escherichia coli. Methods Enzymol. 1975;42:323–329. doi: 10.1016/0076-6879(75)42135-8. [DOI] [PubMed] [Google Scholar]
  23. VISHNIAC W., SANTER M. The thiobacilli. Bacteriol Rev. 1957 Sep;21(3):195–213. doi: 10.1128/br.21.3.195-213.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Valentini G., Iadarola P., Somani B. L., Malcovati M. Two forms of pyruvate kinase in Escherichia coli. A comparison of chemical and molecular properties. Biochim Biophys Acta. 1979 Oct 11;570(2):248–258. doi: 10.1016/0005-2744(79)90145-1. [DOI] [PubMed] [Google Scholar]
  25. Van Dijken J. P., Quayle J. R. Fructose metabolism in four Pseudomonas species. Arch Microbiol. 1977 Sep 28;114(3):281–286. doi: 10.1007/BF00446874. [DOI] [PubMed] [Google Scholar]
  26. Warwick S., Bowen T., McVeigh H., Embley T. M. A phylogenetic analysis of the family Pseudonocardiaceae and the genera Actinokineospora and Saccharothrix with 16S rRNA sequences and a proposal to combine the genera Amycolata and Pseudonocardia in an emended genus Pseudonocardia. Int J Syst Bacteriol. 1994 Apr;44(2):293–299. doi: 10.1099/00207713-44-2-293. [DOI] [PubMed] [Google Scholar]
  27. White P. J., Nairn J., Price N. C., Nimmo H. G., Coggins J. R., Hunter I. S. Phosphoglycerate mutase from Streptomyces coelicolor A3(2): purification and characterization of the enzyme and cloning and sequence analysis of the gene. J Bacteriol. 1992 Jan;174(2):434–440. doi: 10.1128/jb.174.2.434-440.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. de Boer L., Dijkhuizen L., Grobben G., Goodfellow M., Stackebrandt E., Parlett J. H., Whitehead D., Witt D. Amycolatopsis methanolica sp. nov., a facultatively methylotrophic actinomycete. Int J Syst Bacteriol. 1990 Apr;40(2):194–204. doi: 10.1099/00207713-40-2-194. [DOI] [PubMed] [Google Scholar]

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

RESOURCES