1887

Abstract

-Glutamine -fructose-6-phosphate amidotransferase (‘glucosamine synthase’, EC 5.3.1.19) from MRE 600 was purified at least 75-fold. It catalysed the formation of 21·1 μmol glucosamine 6-phosphate (mg protein) in 30 min at 37 °C. Its molecular weight, estimated by gel filtration, was about 90000 and it was inhibited by thiol group reagents. Anticapsin, the C-terminal amino acid of the dipeptide antibiotic bacilysin, and to a lesser extent bacilysin itself, inhibited glucosamine synthase activity. Kinetic studies indicated that the inhibition was non-competitive with respect to fructose 6-phosphate as substrate but partly competitive with respect to -glutamine. Incubation of the enzyme with anticapsin brought about a time-dependent and irreversible inhibition. It is suggested that anticapsin behaves as a glutamine analogue and that a reaction of its epoxide group with a thiol group of glucosamine synthase results in its linkage to the enzyme by a covalent bond.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-131-2-265
1985-02-01
2021-07-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/131/2/mic-131-2-265.html?itemId=/content/journal/micro/10.1099/00221287-131-2-265&mimeType=html&fmt=ahah

References

  1. Atsumi K., Oiwa R., Omura S. 1975; Production of bacillin by Bacillus subtilis sp. strain No. KM-208 and its identity with tetaine (bacilysin). Journal of Antibiotics 28:77–78
    [Google Scholar]
  2. Bates C. J., Handschumacher R. E. 1969; Inactivation and resynthesis of glucosamine-6-phosphate synthetase after treatment with glutamine analogs. Advances in Enzyme Regulation 7:183–204
    [Google Scholar]
  3. Borowski E. 1953; Isolation of pure tetaine from theta strain of Bacillus pumilus. Bulletin of the State Institute of Marine and Tropical Medicine in Gdansk 5:294–306
    [Google Scholar]
  4. Bradford M. M. 1976; A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  5. Buchanan J. M. 1973; The amidotransferases. Advances in Enzymology 39:91–183
    [Google Scholar]
  6. Chmara H., Smulkowski M., Borowski E. 1980; Growth inhibitory effect of amidotransferase inhibition in Candida albicans by epoxypeptides. Drugs under Experimental and Clinical Research 6:7–14
    [Google Scholar]
  7. Chmara H., Woynarowska B., Borowski E. 1981; Epoxypeptide antibiotic tetaine mimics peptide in transport to bacteria. Journal of Antibiotics 34:1608–1612
    [Google Scholar]
  8. Chmara H., Milewski S., Dzieduszycka M., Smulkowski M., Sawlewicz P., Borowski E. 1982; Epoxypeptides - a novel group of metabolic inhibitors in prokaryotic and eukaryotic organisms. Drugs under Experimental and Clinical Research 8:11–12
    [Google Scholar]
  9. Cornish-Bowden A. 1976 Principles of Enzyme Kinetics57–61 London & Boston:: Butterworths;
    [Google Scholar]
  10. Ghosh S., Blumenthal H. J., Davidson E., Roseman S. 1960; Glucosamine metabolism. V. Enzymatic synthesis of glucosamine-6-phosphate. Journal of Biological Chemistry 235:1265–1273
    [Google Scholar]
  11. Kaminski K., Sokolowska J. 1973; The probable identity of bacilysin and tetaine. Journal of Antibiotics 26:184–185
    [Google Scholar]
  12. Kenig M., Abraham E. P. 1976; Antimicrobial activities and antagonists of bacilysin and anticapsin. Journal of General Microbiology 94:37–45
    [Google Scholar]
  13. Kenig M., Vandamme E., Abraham E. P. 1976; The mode of action of bacilysin and anticapsin and biochemical properties of bacilysin-resistant mutants. Journal of General Microbiology 94:46–54
    [Google Scholar]
  14. Kornfeld R. 1967; Studies on l-glutamine: d-fructose-6-phosphate amidotransferase. Journal of Biological Chemistry 242:3135–3141
    [Google Scholar]
  15. Layne E. 1957; Spectrophotometric and turbidimetric methods for measuring proteins. Methods in Enzymology 3:447–454
    [Google Scholar]
  16. Meloche P. 1967; Bromopyruvate inactivation of 2-keto-3-deoxy-6-phosphogluconic aldolase. I. Kinetic evidence for active site specificity. Biochemistry 6:2273–2280
    [Google Scholar]
  17. Mitchell P. 1949; A new technique for stirred aerated culture. Nature London: 164846
    [Google Scholar]
  18. Neuss N., Molloy B. B., Shah R., de la Higuera N. 1970; The structure of anticapsin a new biologically active metabolite of Streptomyces griseo-planus. Biochemical Journal 118:571–575
    [Google Scholar]
  19. Norrman J., Myers R. B., Giddings T. H., Cantino E. C. 1973; Partial purification of l-glutamine : d-fructose-6-phosphate amidotransferase from zoospores of Blastocladiella emersonii. Biochimica et biophysica acta 302:173–177
    [Google Scholar]
  20. Payne J. W. 1972; The characterization of dipeptidases from Escherichia coli. Journal of General Microbiology 71:267–279
    [Google Scholar]
  21. Perry O., Abraham E. P. 1979; Transport and metabolism of bacilysin and other peptides by suspensions of Staphylococcus aureus. Journal of General Microbiology 115:213–221
    [Google Scholar]
  22. Rogers H. J., Lomakina N., Abraham E. P. 1965; Observation on the structure of bacilysin. Biochemical Journal 97:579–586
    [Google Scholar]
  23. Walker J. E., Abraham E. P. 1970; The structure of bacilysin and other products of Bacillus subtilis. Biochemical Journal 118:563–570
    [Google Scholar]
  24. Whitney J. G., Funderburk S. S. 1970; Anticapsin, a new biologically active metabolite. IV. Mechanism of action. Abstracts for the tenth International Congress for Microbiology Mexico City:101 Pérez-Miravete A., Peláez. D.
    [Google Scholar]
  25. Whitney J. G., Funderburk S. S., Westhead J. E., Lively D. H., Solenburg J. M., Denney J. W. 1970; Anticapsin, a new biologically active metabolite. I. Anticapsular screen and assay. Bacteriological Proceedings 7.:
    [Google Scholar]
  26. Winterburn P. J., Phelps C. F. 1971; Purification and some kinetic properties of rat liver glucosamine synthetase. Biochemical Journal 121:701–709
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-131-2-265
Loading
/content/journal/micro/10.1099/00221287-131-2-265
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error