Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Mar;63(3):1189–1191. doi: 10.1128/aem.63.3.1189-1191.1997

Use of 4-Nitrophenoxyacetic Acid for Detection and Quantification of 2,4-Dichlorophenoxyacetic Acid (2,4-D)/(alpha)-Ketoglutarate Dioxygenase Activity in 2,4-D-Degrading Microorganisms

T M Sassanella, F Fukumori, M Bagdasarian, R P Hausinger
PMCID: PMC1389142  PMID: 16535548

Abstract

Purified 2,4-dichlorophenoxyacetic acid (2,4-D)/(alpha)-ketoglutarate dioxygenase (TfdA) was shown to use 4-nitrophenoxyacetic acid (K(infm) = 0.89 (plusmn) 0.04 mM, k(infcat) [catalytic constant] = 540 (plusmn) 10 min(sup-1)), producing intensely yellow 4-nitrophenol. This reagent was used to develop a rapid, continuous, colorimetric assay for the detection of TfdA and analogous activities in 2,4-D-degrading bacterial cells and extracts.

Full Text

The Full Text of this article is available as a PDF (66.0 KB).

Selected References

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

  1. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Don R. H., Weightman A. J., Knackmuss H. J., Timmis K. N. Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4). J Bacteriol. 1985 Jan;161(1):85–90. doi: 10.1128/jb.161.1.85-90.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fukumori F., Hausinger R. P. Alcaligenes eutrophus JMP134 "2,4-dichlorophenoxyacetate monooxygenase" is an alpha-ketoglutarate-dependent dioxygenase. J Bacteriol. 1993 Apr;175(7):2083–2086. doi: 10.1128/jb.175.7.2083-2086.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fukumori F., Hausinger R. P. Purification and characterization of 2,4-dichlorophenoxyacetate/alpha-ketoglutarate dioxygenase. J Biol Chem. 1993 Nov 15;268(32):24311–24317. [PubMed] [Google Scholar]
  5. Fulthorpe R. R., McGowan C., Maltseva O. V., Holben W. E., Tiedje J. M. 2,4-Dichlorophenoxyacetic acid-degrading bacteria contain mosaics of catabolic genes. Appl Environ Microbiol. 1995 Sep;61(9):3274–3281. doi: 10.1128/aem.61.9.3274-3281.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fulthorpe R. R., Rhodes A. N., Tiedje J. M. Pristine soils mineralize 3-chlorobenzoate and 2,4-dichlorophenoxyacetate via different microbial populations. Appl Environ Microbiol. 1996 Apr;62(4):1159–1166. doi: 10.1128/aem.62.4.1159-1166.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harker A. R., Olsen R. H., Seidler R. J. Phenoxyacetic acid degradation by the 2,4-dichlorophenoxyacetic acid (TFD) pathway of plasmid pJP4: mapping and characterization of the TFD regulatory gene, tfdR. J Bacteriol. 1989 Jan;171(1):314–320. doi: 10.1128/jb.171.1.314-320.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. King R. J., Short K. A., Seidler R. J. Assay for detection and enumeration of genetically engineered microorganisms which is based on the activity of a deregulated 2,4-dichlorophenoxyacetate monooxygenase. Appl Environ Microbiol. 1991 Jun;57(6):1790–1792. doi: 10.1128/aem.57.6.1790-1792.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Michel L. O., Sandkvist M., Bagdasarian M. Specificity of the protein secretory apparatus: secretion of the heat-labile enterotoxin B subunit pentamers by different species of gram- bacteria. Gene. 1995 Jan 11;152(1):41–45. doi: 10.1016/0378-1119(94)00691-k. [DOI] [PubMed] [Google Scholar]
  10. Perkins E. J., Gordon M. P., Caceres O., Lurquin P. F. Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4. J Bacteriol. 1990 May;172(5):2351–2359. doi: 10.1128/jb.172.5.2351-2359.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Perkins E. J., Lurquin P. F. Duplication of a 2,4-dichlorophenoxyacetic acid monooxygenase gene in Alcaligenes eutrophus JMP134(pJP4). J Bacteriol. 1988 Dec;170(12):5669–5672. doi: 10.1128/jb.170.12.5669-5672.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Streber W. R., Timmis K. N., Zenk M. H. Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134. J Bacteriol. 1987 Jul;169(7):2950–2955. doi: 10.1128/jb.169.7.2950-2955.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES