Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1977 Jun 15;164(3):579–587. doi: 10.1042/bj1640579

Bacterial catabolism of threonine. Threonine degradation initiated by l-threonine hydrolyase (deaminating) in a species of Corynebacterium

Stephen C Bell 1,*, John M Turner 1
PMCID: PMC1164834  PMID: 16743051

Abstract

1. Three bacterial isolates capable of growth on l-threonine medium only when supplemented with branched-chain amino acids, and possessing high l-threonine dehydratase activity, were examined to elucidate the catabolic route for the amino acid. 2. Growth, manometric, radiotracer and enzymic experiments indicated that l-threonine was catabolized by initial deamination to 2-oxobutyrate and thence to propionate. No evidence was obtained for the involvement of l-threonine 3-dehydrogenase or l-threonine aldolase in threonine catabolism. 3. l-Threonine dehydratase of Corynebacterium sp. F5 (N.C.I.B. 11102) was partially purified and its kinetic properties were examined. The enzyme exhibited a sigmoid kinetic response to substrate concentration. The concentration of substrate giving half the maximum velocity, [S0.5], was 40mm and the Hill coefficient (h) was 2.0. l-Isoleucine inhibited enzyme activity markedly, causing 50% inhibition at 60μm, but did not affect the Hill constant. At the fixed l-threonine concentration of 10mm, the effect of l-valine was biphasic, progressive activation occurring at concentrations up to 2mm-l-valine, but was abolished by higher concentrations. Substrate-saturation plots for the l-valine-activated enzyme exhibited normal Michaelis–Menten kinetics with a Hill coefficient (h) of 1.0. The kinetic properties of the enzyme were thus similar to those of the `biosynthetic' isoenzyme from Rhodopseudomonas spheroides rather than those of the enteric bacteria. 4. The synthesis of l-threonine dehydratase was constitutive and was not subject to multivalent repression by l-isoleucine or other branched-chain amino acids either singly or in combination. 5. The catabolism of l-threonine, apparently initiated by a `biosynthetic' l-threonine dehydratase in the isolates studied, depended on the concomitant catabolism of branched-chain amino acids. The biochemical basis of this dependence appeared to lie in the further catabolism of 2-oxobutyrate by enzymes which required branched-chain 2-oxo acids for their induction.

Full text

PDF
579

Selected References

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

  1. Barrtt G. J. Biosynthesis of isoleucine and valine in Rhodopseudomonas spheroides: regulation of threonine deaminase activity. J Bacteriol. 1971 Mar;105(3):718–721. doi: 10.1128/jb.105.3.718-721.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell S. C., Turner J. M. Bacterial catabolism of threonine. Threonine degradation initiated by L-threonine-NAD+ oxidoreductase. Biochem J. 1976 May 15;156(2):449–458. doi: 10.1042/bj1560449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bell S. C., Turner J. M. L-Threonine catabolism via aminoacetone: a search for a pathway in bacteria. Biochem Soc Trans. 1976;4(3):497–500. doi: 10.1042/bst0040497. [DOI] [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. FREUNDLICH M., BURNS R. O., UMBARGER H. E. Control of isoleucine, valine, and leucine biosynthesis. I. Multivalent repression. Proc Natl Acad Sci U S A. 1962 Oct 15;48:1804–1808. doi: 10.1073/pnas.48.10.1804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gupta Y. P. Threonine deaminase (dehydratase) in Azotobacter chroococcum. Enzymologia. 1971 Aug 31;41(2):91–98. [PubMed] [Google Scholar]
  7. Hall T. C., Cocking E. C. High-efficiency liquid-scintillation counting of 14C-labelled material in aqueous solution and determination of specific activity of labelled proteins. Biochem J. 1965 Sep;96(3):626–633. doi: 10.1042/bj0960626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hayakawa T., Hirashima M., Ide S., Hamada M., Okabe K., Koike M. Mammalian alpha-keto acid dehydrogenase complexes. I. Isolation, purification, and properties of pyruvate dehydrogenase complex of pig heart muscle. J Biol Chem. 1966 Oct 25;241(20):4694–4699. [PubMed] [Google Scholar]
  9. Higgins I. J., Pickard M. A., Turner J. M. Aminoacetone formation and utilization by pseudomonads grown on DL-1-aminopropan-2-ol. J Gen Microbiol. 1968 Nov;54(1):105–114. doi: 10.1099/00221287-54-1-105. [DOI] [PubMed] [Google Scholar]
  10. Hill A. V. The Combinations of Haemoglobin with Oxygen and with Carbon Monoxide. I. Biochem J. 1913 Oct;7(5):471–480. doi: 10.1042/bj0070471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KORNBERG H. L. The metabolism of C2 compounds in micro-organisms. I. The incorporation of [2-14C] acetate by Pseudomonas fluorescens, and by a Corynebacterium, grown on ammonium acetate. Biochem J. 1958 Mar;68(3):535–542. doi: 10.1042/bj0680535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Katsunuma T., Elsässer S., Holzer H. Purification and some properties of threonine dehydratase from yeast. Eur J Biochem. 1971 Dec 22;24(1):83–87. doi: 10.1111/j.1432-1033.1971.tb19657.x. [DOI] [PubMed] [Google Scholar]
  13. Kumagai H., Nagate T., Yoshida H., Yamada H. Threonine aldolase from Candida humicola. II. Purification, crystallization and properties. Biochim Biophys Acta. 1972 Mar 8;258(3):779–790. doi: 10.1016/0005-2744(72)90179-9. [DOI] [PubMed] [Google Scholar]
  14. Marshall V. D., Sokatch J. R. Regulation of valine catabolism in Pseudomonas putida. J Bacteriol. 1972 Jun;110(3):1073–1081. doi: 10.1128/jb.110.3.1073-1081.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Massey L. K., Conrad R. S., Sokatch J. R. Regulation of leucine catabolism in Pseudomonas putida. J Bacteriol. 1974 Apr;118(1):112–120. doi: 10.1128/jb.118.1.112-120.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Massey L. K., Sokatch J. R., Conrad R. S. Branched-chain amino acid catabolism in bacteria. Bacteriol Rev. 1976 Mar;40(1):42–54. doi: 10.1128/br.40.1.42-54.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGilvray D., Morris J. G. Utilization of L-threonine by a species of Arthrobacter. A novel catabolic role for "aminoacetone synthase". Biochem J. 1969 May;112(5):657–671. doi: 10.1042/bj1120657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miyajima R., Shiio I. Regulation of aspartate family amino acid biosynthesis in Brevibacterium flavum. VI. Effects of isoleucine and valine on threonine dehydratase activity and its formation. J Biochem. 1972 Jun;71(6):951–960. doi: 10.1093/oxfordjournals.jbchem.a129866. [DOI] [PubMed] [Google Scholar]
  19. Morris J. G. Utilization of L-threnonine by a pseudomonad: a catabolic role for L-threonine aldolase. Biochem J. 1969 Nov;115(3):603–605. doi: 10.1042/bj1150603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ning C., Gest H. Regulation of L-isoleucine biosynthesis in the photosynthetic bacterium rhodospirillum rubrum. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1823–1827. doi: 10.1073/pnas.56.6.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tokushige M., Hayaishi O. Threonine metabolism and its regulation in Clostridium tetanomorphum. J Biochem. 1972 Aug;72(2):469–477. doi: 10.1093/oxfordjournals.jbchem.a129923. [DOI] [PubMed] [Google Scholar]
  22. UMBARGER H. E., BROWN B. Threonine deamination in Escherichia coli. II. Evidence for two L-threonine deaminases. J Bacteriol. 1957 Jan;73(1):105–112. doi: 10.1128/jb.73.1.105-112.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Umbarger H. E. Threonine deaminases. Adv Enzymol Relat Areas Mol Biol. 1973;37:349–395. doi: 10.1002/9780470122822.ch6. [DOI] [PubMed] [Google Scholar]
  24. Willetts A. J., Turner J. M. L-Threonine acetaldehyde-lyase in a strain of Bacillus subtilis. Biochim Biophys Acta. 1971 Oct;252(1):105–110. doi: 10.1016/0304-4165(71)90097-3. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES