Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 May;114(2):556–562. doi: 10.1128/jb.114.2.556-562.1973

Purification and Properties of Threonine Deaminase from the X-1 Isolate of the Genus Thermus

Edward H Higa a,1, Robert F Ramaley a,2
PMCID: PMC251809  PMID: 4706189

Abstract

Threonine deaminase (l-threonine dehydratase EC 4.2.1.16) has been partially purified from a new extreme thermophilic bacterium, Thermus X-1, which is similar to T. aquaticus YT-1. The threonine deaminase of strain X-1 has a maximal rate of reaction at 85 to 90 C and is more thermostable than the threonine deaminase from mesophilic bacteria. The enzyme has an apparent molecular weight of 100,000 to 115,000, a Km for l-threonine of 14 mM, a pH optimum of 8.0, and like other threonine deaminases also catalyzes the deamination of serine. However the Thermus X-1 threonine deaminase does not show a strong feedback inhibition by isoleucine. It is suggested that the regulation of the biosynthesis of isoleucine in this extreme theromophile may resemble that reported in Rodospirillum rubrum.

Full text

PDF
556

Selected References

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

  1. Brock T. D., Freeze H. Thermus aquaticus gen. n. and sp. n., a nonsporulating extreme thermophile. J Bacteriol. 1969 Apr;98(1):289–297. doi: 10.1128/jb.98.1.289-297.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brock T. D. Life at high temperatures. Evolutionary, ecological, and biochemical significance of organisms living in hot springs is discussed. Science. 1967 Nov;158(3804):1012–1019. doi: 10.1126/science.158.3804.1012. [DOI] [PubMed] [Google Scholar]
  3. Burns R. O., Zarlengo M. H. Threonine deaminase from Salmonella typhimurium. I. Purification and properties. J Biol Chem. 1968 Jan 10;243(1):178–185. [PubMed] [Google Scholar]
  4. Datta P. Purification and feedback control of threonine deaminase activity of Rhodopseudomonas spheroides. J Biol Chem. 1966 Dec 25;241(24):5836–5844. [PubMed] [Google Scholar]
  5. Datta P. Regulation of branched biosynthetic pathways in bacteria. Science. 1969 Aug 8;165(3893):556–562. doi: 10.1126/science.165.3893.556. [DOI] [PubMed] [Google Scholar]
  6. Desai I. D., Polglase W. J. Threonine dehydratase of streptomycin-dependent Escherichia coli K-12. Biochim Biophys Acta. 1966 Mar 21;114(3):642–644. doi: 10.1016/0005-2787(66)90116-x. [DOI] [PubMed] [Google Scholar]
  7. Feldberg R. S., Datta P. L-threonine deaminase of Rhodospirillum rubrum. Purification and characterization. Eur J Biochem. 1971 Aug 16;21(3):438–446. doi: 10.1111/j.1432-1033.1971.tb01490.x. [DOI] [PubMed] [Google Scholar]
  8. Feldberg R. S., Datta P. Threonine deaminase: a novel activity stain on polyacrylamide gels. Science. 1970 Dec 25;170(3965):1414–1415. doi: 10.1126/science.170.3965.1414. [DOI] [PubMed] [Google Scholar]
  9. Freeze H., Brock T. D. Thermostable aldolase from Thermus aquaticus. J Bacteriol. 1970 Feb;101(2):541–550. doi: 10.1128/jb.101.2.541-550.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Good N. E., Winget G. D., Winter W., Connolly T. N., Izawa S., Singh R. M. Hydrogen ion buffers for biological research. Biochemistry. 1966 Feb;5(2):467–477. doi: 10.1021/bi00866a011. [DOI] [PubMed] [Google Scholar]
  11. Hatfield G. W., Burns R. O. Ligand-induced maturation of threonine deaminase. Science. 1970 Jan 2;167(3914):75–76. doi: 10.1126/science.167.3914.75. [DOI] [PubMed] [Google Scholar]
  12. Hatfield G. W., Umbarger H. E. Threonine deaminase from Bacillus subtilis. I. Purification of the enzyme. J Biol Chem. 1970 Apr 10;245(7):1736–1741. [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Leitzmann C., Bernlohr R. W. Threonine dehydratase of Bacillus licheniformis. I. Purification and properties. Biochim Biophys Acta. 1968 Feb 5;151(2):449–460. doi: 10.1016/0005-2744(68)90113-7. [DOI] [PubMed] [Google Scholar]
  15. Ramaley R. F., Hixson J. Isolation of a nonpigmented, thermophilic bacterium similar to Thermophilic bacterium similar to Thermus aquaticus. J Bacteriol. 1970 Aug;103(2):527–528. doi: 10.1128/jb.103.2.527-528.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Reisfeld R. A., Small P. A., Jr Electrophoretic heterogeneity of polypeptide chains of specific antibodies. Science. 1966 May 27;152(3726):1253–1255. doi: 10.1126/science.152.3726.1253. [DOI] [PubMed] [Google Scholar]
  17. Siegelman H. W., Wieczorek G. A., Turner B. C. Preparation of calcium phosphate for protein chromatography. Anal Biochem. 1965 Dec;13(3):402–404. doi: 10.1016/0003-2697(65)90332-5. [DOI] [PubMed] [Google Scholar]
  18. Singleton R., Jr, Kimmel J. R., Amelunxen R. E. The amino acid composition and other properties of thermostable glyceraldehyde 3-phosphate dehydrogenase from Bacillus stearothermophilus. J Biol Chem. 1969 Mar 25;244(6):1623–1630. [PubMed] [Google Scholar]
  19. Thomas D. A., Kuramitsu H. K. Biosynthetic L-threonine deaminase from Bacillus stearothermophilus. I. Catalytic and regulatory properties. Arch Biochem Biophys. 1971 Jul;145(1):96–104. doi: 10.1016/0003-9861(71)90014-2. [DOI] [PubMed] [Google Scholar]
  20. UMBARGER H. E. Evidence for a negative-feedback mechanism in the biosynthesis of isoleucine. Science. 1956 May 11;123(3202):848–848. doi: 10.1126/science.123.3202.848. [DOI] [PubMed] [Google Scholar]
  21. WILLIAMS D. E., REISFELD R. A. DISC ELECTROPHORESIS IN POLYACRYLAMIDE GELS: EXTENSION TO NEW CONDITIONS OF PH AND BUFFER. Ann N Y Acad Sci. 1964 Dec 28;121:373–381. doi: 10.1111/j.1749-6632.1964.tb14210.x. [DOI] [PubMed] [Google Scholar]
  22. WOOD W. A., GUNSALUS I. C. Serine and threonine desaminaes of Escherichia coli; activators for a cell-free enzyme. J Biol Chem. 1949 Nov;181(1):171–182. [PubMed] [Google Scholar]
  23. Zeikus J. G., Brock T. D. Protein synthesis at high temperatures: aminoacylation of tRNA. Biochim Biophys Acta. 1971 Feb 11;228(3):736–745. doi: 10.1016/0005-2787(71)90739-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES