1887

Abstract

has two chromatographically distinct forms of glutamate dehydrogenase (GDH): one GDH utilizes NAD as coenzyme, the other uses NADP. The intracellular level of both GDHs is strongly regulated by the nitrogen source in the growth medium. NADP-dependent GDH was purified to homogeneity from crude extracts of The of the native enzyme was determined to be 200000 by size-exclusion high-performance liquid chromatography whereas after sodium dodecyl sulphate-polyacrylamide gel electrophoresis one major band of 49000 was found, suggesting that the enzyme is a tetramer. The enzyme was highly specific for the substrates 2-oxoglutarate and -glutamate, and required NADP, which could not be replaced by NAD, as a cofactor. The pH optimum was 9·2 for oxidative deamination of glutamate and 8·4 for reductive amination of 2-oxoglutarate. The Michaelis constants ( ) were 28·6 m for -glutamate and 0·12 m for NADP. values for reductive amination were 1·54 m for 2-oxoglutarate, 0·07 m for NADPH and 30·8 m for NH . The enzyme activity was significantly reduced by adenine nucleotides, particularly ATP.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-12-3311
1989-12-01
2021-10-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/12/mic-135-12-3311.html?itemId=/content/journal/micro/10.1099/00221287-135-12-3311&mimeType=html&fmt=ahah

References

  1. Aharonowitz Y., Friedrich C. G. 1980; Alanine dehydrogenase of the β-lactam antibiotic producer Streptomyces clavuligerus.. Archives of Microbiology 125:137–142
    [Google Scholar]
  2. Bonete M. J., Camacho M. L., Cadenas E. 1987; A new glutamate dehydrogenase from Halobacterium halobium with different coenzyme specificity.. International Journal of Biochemistry 19:1149–1155
    [Google Scholar]
  3. BraÑa A. F., Paiva N., Demain A. L. 1986; Pathways and regulation of ammonium assimilation in Streptomyces clavuligerus.. Journal of General Microbiology 132:1305–1317
    [Google Scholar]
  4. Coulton J. W., Kapoor M. 1972; Studies on the kinetics and regulation of glutamate dehydrogenase of Salmonella typhimurium.. Canadian Journal of Microbiology 19:439–450
    [Google Scholar]
  5. Ferguson A. R., Sims A. P. 1971; Inactivation in vivo of glutamine synthetase and NAD-specific glutamate dehydrogenase: its role in the regulation of glutamine synthesis in yeasts.. Journal of General Microbiology 69:423–427
    [Google Scholar]
  6. Fisher S. H. 1988; Nitrogen assimilation in Streptomyces.. In Biology of Actinomycetes pp. 47–51 Okami Y., Beppu T., Ogawaza H. Edited by Tokyo: Japan Scientific Societies Press;
    [Google Scholar]
  7. GrÄfe U., Bocker H., Thrum H. 1977; Regulative influence of o-aminobenzoic acid on the biosynthesis of nourseothricin in cultures Streptomyces noursei JA 3890b.. Zeitschrift für allgemeine Mikrobiologie 17:201–209
    [Google Scholar]
  8. Itoh N., Morikawa R. 1983; Crystallization and properties of l-alanine dehydrogenase from Streptomyces phaeochromogenes.. Agricultural and Biological Chemistry 47:2511–2519
    [Google Scholar]
  9. Kew O. M., Woolfolk C. A. 1970; Preparation of glutamate dehydrogenase from Micrococcus aerogenes.. Biochemical and Biophysical Research Communications 39:1126–1133
    [Google Scholar]
  10. Kramer J. 1970; NAD- and NADP-dependent glutamate dehydrogenase in Hydrogenomonas H 16.. Archiv für Mikrobiologie 71:226–234
    [Google Scholar]
  11. Kumar S., Nicholas D.J.D. 1984; Purification, properties and regulation of glutamine synthetase from Nitrobacter agilis.. Journal of General Microbiology 130:959–966
    [Google Scholar]
  12. Leicht W., Weber M. M., Eisenberg H. 1978; Purification and characterization of glutamate dehydrogenase from Halobacterium of the Dead Sea.. Biochemistry 17:4004–4010
    [Google Scholar]
  13. Le’John H. B., Suzuki I., Wright J. A. 1968; Glutamate dehydrogenase of Thiobacillus novellus. Kinetic properties and a possible control mechanism.. Journal of Biological Chemistry 243:118–128
    [Google Scholar]
  14. Navarrete R. M., Vara J. A., Hutchinson C. R. 1989; Inducible l-valine dehydrogenase in Streptomyces coelicolor and other Streptomyces.. Journal of Bacteriology (in the Press)
    [Google Scholar]
  15. Ōmura S., Tanaka Y., Mamada H., Masuma R. 1983; Ammonium ion suppresses the biosynthesis of tylosin aglycone by interference with valine catabolism in Streptomyces fradiae.. Journal of Antibiotics 36:1792–1794
    [Google Scholar]
  16. Phibbs P. V., Bernlohr R. W. 1971; Purification, properties and regulation of glutamic dehydrogenase of Bacillus licheniformis.. Journal of Bacteriology 106:375–385
    [Google Scholar]
  17. Priestley N. D., Robinson J. A. 1989; Purification and catalytic properties of l-valine dehydrogenase from Streptomyces cinnamonensis.. Biochemical Journal 261:853–861
    [Google Scholar]
  18. Shapiro S., Vining L. C. 1983; Nitrogen metabolism and chloramphenicol production in Streptomyces venezuelae.. Canadian Journal of Microbiology 29:1706–1714
    [Google Scholar]
  19. VanČura A., Řezanka T., MarŠÁlek J., VanČurovÁ I., KŘiŠŤan V., BasaŘovÁ G. 1987; Effect of ammonium ions on the composition of fatty acids in Streptomyces fradiae,producer of tylosin.. FEMS Microbiology Letters 48:357–360
    [Google Scholar]
  20. VanČura A., VanČurovÁ I., Volc J., Fussey S.P.M., Flieger M., NeuŽil J., MarŠÁlek J., BĚhal V. 1988a; Valine dehydrogenase from Streptomyces fradiae: purification and properties.. Journal of General Microbiology 134:3213–3219
    [Google Scholar]
  21. VanČura A., Řezanka T., MarŠÁlek J., Melzoch K., BasaŘovÁ G., KŘiŠŤan V. 1988b; Metabolism of l-threonine and fatty acids and tylosin biosynthesis in Streptomyces fradiae.. FEMS Microbiology Letters 49:411–415
    [Google Scholar]
  22. VanČura A., VanČurovÁ I., KopeckÝ J., MarŠÁlek J., CikÁnek D., BasaŘovÁ G., KŘiŠŤan V. 1989a; Regulation of branched-chain amino acid biosynthesis in Streptomyces fradiae, a producer of tylosin.. Archives of Microbiology 151:537–540
    [Google Scholar]
  23. VanČura A., VanČurovÁ I., Volc J., Jones S.K.T., Flieger M., BasaŘovÁ G., BĚhal V. 1989b; Alanine dehydrogenase from Streptomyces fradiae.Purification and properties.. European Journal of Biochemistry 179:221–227
    [Google Scholar]
  24. VanČurovÁ I., VanČura A., Volc J., NeuŽil J., Flieger M., BasaŘovÁ G., BĚhal V. 1988a; Isolation and characterization of valine dehydrogenase from Streptomyces aureofaciens.. Journal of Bacteriology 170:5192–5196
    [Google Scholar]
  25. VanČurovÁ I., VanČura A., Volc J., NeuŽil J., Flieger M., BasaŘovÁ G., BĚhal V. 1988b; Purification and partial characterization of alanine dehydrogenase from Streptomyces aureofaciens.. Archives of Microbiology 150:438–440
    [Google Scholar]
  26. VanČurovÁ I., VanČura A., Volc J., NeuŽil J., Flieger M., BĚhal V. 1988c; NAD-dependent l-amino acid dehydrogenases from Streptomyces aureofaciens.. 14th International Congress of Biochemistry, Prague, Abstract book p. 166:
    [Google Scholar]
  27. VanČurovÁ I., Volc J., Flieger M., NeuŽil J., NovotnÁ J., Vlach J., BĚhal V. 1988d; Isolation of pure anhydrotetracycline oxygenase from Streptomyces aureofaciens.. Biochemical Journal 253:263–267
    [Google Scholar]
  28. Whitaker J. R., Granum P. E. 1980; An absolute method for protein determination based on difference in absorbance at 235 and 280 nm.. Analytical Biochemistry 109:156–159
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-12-3311
Loading
/content/journal/micro/10.1099/00221287-135-12-3311
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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