1887

Abstract

SUMMARY: The 2-hydroxy-6-oxohepta-2,4-dienoate (HOD) hydrolase encoded by the TOL plasmid pWW0 from mt-2 (PaWl) was purified to homogeneity. It has an of 65000 and is dissociated by SDS into two subunits of equal size. Alanine was the only N-terminal residue detected, and each subunit contained one cysteine thiol group. The pH optimum for activity and for enzyme stability was around 7·5, whereas the isoelectric point was 4·7. Only the products of catechol 2,3-oxygenase action on linear chain alkylcatechols and 4-chlorocatechol served as substrates. Kinetic measurements showed that the higher activity against the ketone substrates (from 3-substituted catechols) over aldehyde substrates (from 4-substituted catechols) was the result of higher values rather than lower values. Antisera prepared against this purified HOD hydrolase were shown by Ouchterlony double diffusion and inhibition studies to be related to the HOD hydrolase from phenol-grown strain U (NCIB 10015) in which the enzyme is chromosomally encoded.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-132-3-717
1986-03-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/132/3/mic-132-3-717.html?itemId=/content/journal/micro/10.1099/00221287-132-3-717&mimeType=html&fmt=ahah

References

  1. Bayly R. C., Di Berardino D. 1978; Purification and properties of 2-hydroxy-6-oxo-2,4-heptadien-oate hydrolase from two strains of Pseudomonas putida. Journal of Bacteriology 134:30–37
    [Google Scholar]
  2. Bayly R. C., Dagley S., Gibson D. T. 1966; The metabolism of cresols by species of Pseudomonas. Biochemical Journal 101:293–301
    [Google Scholar]
  3. Benesch R., Benesch R. E. 1962; Determination of SH groups in proteins. Methods of Biochemical Analysis 10:43–70
    [Google Scholar]
  4. Blackburn W. H., Barker L., Catchpole J. R., Hollingworth N. W. 1955; Gas Council Research Communications, GC24. An Investigation into the Composition of Ammoniacal Liquor: Continuous Retort Liquor.
    [Google Scholar]
  5. Conway-Jacobs A., Lewin M. L. 1971; Isoelectric focussing in acrylamide gels. Use of amphoteric dyes as internal markers for determination of isoelectric points. Analytical Biochemistry 43:394–400
    [Google Scholar]
  6. Davis B. J. 1964; Disc electrophoresis. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121:404–427
    [Google Scholar]
  7. Eisenthal R., Cornish-Bowden A. 1974; The direct linear plot: a new graphical procedure for estimating kinetic parameters. Biochemical Journal 139:715–720
    [Google Scholar]
  8. Evans R. A., Thomas A. J., Axford R. F. E., Siriwardine J. A. S., Robins A. J. 1966; Modification of the Technicon five-column amino acid analyser and its application to biological materials. In Proceedings of Vth International Symposium on Automation in Analytical Chemistry, London, 1965, vol. 2, New York: Technicon Corporation. 2 pp:687–691
    [Google Scholar]
  9. Hirs C. H. W. 1966; Determination of cystine as cysteic acid. Methods in Enzymology 11:59–62
    [Google Scholar]
  10. Inouye S., Nakazawa A., Nakazawa T. 1981; Molecular cloning of gene xylS of the TOL plasmid: evidence for positive regulation of the xylDEFG operon by xylS. Journal of Bacteriology 148:413–418
    [Google Scholar]
  11. Liu T.-Y, Chang Y. H. 1971; Hydrolysis of proteins with p-toluene sulphonic acid: determination of tryptophan. Journal of Biological Chemistry 246:2842–2848
    [Google Scholar]
  12. Murray K., Duggleby C. J., Sala-Trepat J.-M., Williams P. A. 1972; The metabolism of benzoate and the methylbenzoates via the meta cleavage pathway by Pseudomonas arvilla mt-2. European Journal of Biochemistry 28:301–310
    [Google Scholar]
  13. Radola B. J. 1973; Isoelectric focussing in layers of granulated gels. I. Thin layer isoelectric focussing of proteins. Biochimica et biophysica acta 295:412–428
    [Google Scholar]
  14. Sala-Trepat J.-M., Evans W. C. 1971; The meta cleavage of catechol by Azotobacter species. European Journal of Biochemistry 20:400–413
    [Google Scholar]
  15. Sala-Trepat J.-M., Murray K., Williams P. A. 1971; The meta cleavage of catechol by Azotobacter species. European Journal of Biochemistry 20:400–413
    [Google Scholar]
  16. Singh T. N., Paleg L. G., Aspinall D. 1973; Stress metabolism. Nitrogen metabolism and growth of barley plant during water stress. Australian Journal of Biological Sciences 26:45–56
    [Google Scholar]
  17. Troll W., Lindsey J. 1955; A photometric method for the determination of proline. Journal of Biological Chemistry 215:655–660
    [Google Scholar]
  18. Weber K., Osborn M. 1969; The reliability of molecular weight determination by dodecyl sulphate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
  19. Weiner A. M., Platt T., Weber K. 1972; Aminoacid sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. Journal of Biological Chemistry 247:2371–2381
    [Google Scholar]
  20. Wigmore G. J., Bayly R. C., Di Berardino D. 1974; Pseudomonas putida mutants defective in the metabolism of meta fission product of catechol and its methyl analogues. Journal of Bacteriology 120:31–37
    [Google Scholar]
  21. Wilkinson G. N. 1961; Statistical estimations in enzyme kinetics. Biochemical Journal 80:324–332
    [Google Scholar]
  22. Worsey M. J., Williams P. A. 1975; Metabolism of toluene and xylenes by Pseudomonas putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. Journal of Bacteriology 124:7–13
    [Google Scholar]
  23. Wrigley C. W. 1969; Instrument applications no. 29. London: Shandon Scientific Co. Analytical Fractionation of Proteins According to Isoelectric Point by Gel Electrofocussing.
    [Google Scholar]
  24. Zahler W. L., Cleland W. W. 1968; A specific and sensitive assay for disulphides. Journal of Biological Chemistry 243:716–719
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-132-3-717
Loading
/content/journal/micro/10.1099/00221287-132-3-717
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