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PMID: 8449889 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Cloning and characterization of Pseudomonas sp. strain DNT genes for 2,4-dinitrotoluene degradation.

Journal of bacteriology ·Vol. 175 ·No. 6 ·1993-03-00 ·Pages 1831-7

Suen WC, Spain JC

Abstract

The degradation of 2,4-dinitrotoluene (DNT) by Pseudomonas sp. strain DNT is initiated by a dioxygenase attack to yield 4-methyl-5-nitrocatechol (MNC) and nitrite. Subsequent oxidation of MNC by a monooxygenase results in the removal of the second molecule of nitrite, and further enzymatic reactions lead to ring fission. Initial studies on the molecular basis of DNT degradation in this strain revealed the presence of three plasmids. Mitomycin-derived mutants deficient in either DNT dioxygenase only or DNT dioxygenase and MNC monooxygenase were isolated. Plasmid profiles of mutant strains suggested that the mutations resulted from deletions in the largest plasmid. Total plasmid DNA partially digested by EcoRI was cloned into a broad-host-range cosmid vector, pCP13. Recombinant clones containing genes encoding DNT dioxygenase, MNC monooxygenase, and 2,4,5-trihydroxytoluene oxygenase were characterized by identification of reaction products and the ability to complement mutants. Subcloning analysis suggests that the DNT dioxygenase is a multicomponent enzyme system and that the genes for the DNT pathway are organized in at least three different operons.

Related Genes
DNT
MeSH Terms
Cloning, Molecular Cosmids Dinitrobenzenes/metabolism Genes, Bacterial Mutation Oxygenases/genetics,metabolism Phenotype Plasmids Pseudomonas/genetics,metabolism Restriction Mapping
Chemicals
Dinitrobenzenes 2,4-dinitrotoluene Oxygenases 2,4-dinitrotoluene dioxygenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Suen W C
Air Force Civil Engineering Support Agency, Tyndall Air Force Base, Florida 32403-6001.
Spain J C
References (38)
38 references, click to expand
  1. Utilization of anthranilic and nitrobenzoic acids by Nocardia opaca and a flavobacterium.
    J Gen Microbiol. 1966 Feb;42(2):219-35 PMID: 5912587
  2. Dinitrotoluene: acute toxicity, oncogenicity, genotoxicity, and metabolism.
    Crit Rev Toxicol. 1984;13(3):217-34 PMID: 6386342
  3. Plasmid gene organization: naphthalene/salicylate oxidation.
    Proc Natl Acad Sci U S A. 1982 Feb;79(3):874-8 PMID: 6278499
  4. Rapid and efficient cosmid cloning.
    Nucleic Acids Res. 1981 Jul 10;9(13):2989-98 PMID: 6269067
  5. Genetic and physical map of the 2,4-dichlorophenoxyacetic acid-degradative plasmid pJP4.
    J Bacteriol. 1985 Jan;161(1):466-8 PMID: 3968031
  6. Enzymatic oxidation of p-nitrophenol.
    Biochem Biophys Res Commun. 1979 May 28;88(2):634-41 PMID: 37837
  7. Toluene dioxygenase: a multicomponent enzyme system.
    Biochem Biophys Res Commun. 1977 Sep 9;78(1):401-10 PMID: 907688
  8. Cloning and expression in Escherichia coli of Acinetobacter calcoaceticus genes for benzoate degradation.
    J Bacteriol. 1987 Dec;169(12):5496-503 PMID: 2824437
  9. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants.
    Gene. 1982 Jun;18(3):289-96 PMID: 6290332
  10. Biotransformation of nitrobenzene by bacteria containing toluene degradative pathways.
    Appl Environ Microbiol. 1991 Nov;57(11):3156-62 PMID: 1781679
  11. Oxidation of nitrotoluenes by toluene dioxygenase: evidence for a monooxygenase reaction.
    Appl Environ Microbiol. 1992 Aug;58(8):2643-8 PMID: 1514810
  12. Identification of biotransformation products from 2,4-dinitrotoluene.
    Appl Environ Microbiol. 1978 May;35(5):945-8 PMID: 566083
  13. A complementation analysis of the restriction and modification of DNA in Escherichia coli.
    J Mol Biol. 1969 May 14;41(3):459-72 PMID: 4896022
  14. Catabolism of 1,3-dinitrobenzene by Rhodococcus sp. QT-1.
    Arch Microbiol. 1991;157(1):76-9 PMID: 1814279
  15. Nitroaromatics Are Substrates for the TOL Plasmid Upper-Pathway Enzymes.
    Appl Environ Microbiol. 1992 Jan;58(1):415-7 PMID: 16348637
  16. Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway.
    Proc Natl Acad Sci U S A. 1981 Dec;78(12):7458-62 PMID: 6950388
  17. Separation and partial characterization of the enzymes of the toluene-4-monooxygenase catabolic pathway in Pseudomonas mendocina KR1.
    J Bacteriol. 1991 May;173(9):3017-20 PMID: 2019564
  18. A multiple plasmid-containing Escherichia coli strain: convenient source of size reference plasmid molecules.
    Plasmid. 1978 Jun;1(3):417-20 PMID: 372973
  19. Cloning and characterization of plasmid-encoded genes for the degradation of 1,2-dichloro-, 1,4-dichloro-, and 1,2,4-trichlorobenzene of Pseudomonas sp. strain P51.
    J Bacteriol. 1991 Jan;173(1):6-15 PMID: 1987135
  20. Characterization of five genes in the upper-pathway operon of TOL plasmid pWW0 from Pseudomonas putida and identification of the gene products.
    J Bacteriol. 1989 Sep;171(9):5048-55 PMID: 2549010
  21. Primary structure of xylene monooxygenase: similarities to and differences from the alkane hydroxylation system.
    J Bacteriol. 1991 Mar;173(5):1690-5 PMID: 1999388
  22. Metabolism of arylsulphonates by micro-organisms.
    Biochem J. 1968 Feb;106(4):859-77 PMID: 5637368
  23. Biodegradation of 2,4-dinitrotoluene by a Pseudomonas sp.
    Appl Environ Microbiol. 1991 Nov;57(11):3200-5 PMID: 1781682
  24. Oxidation of naphthalene by a multicomponent enzyme system from Pseudomonas sp. strain NCIB 9816.
    J Bacteriol. 1982 Mar;149(3):948-54 PMID: 7037744
  25. Genetic basis of the biodegradation of salicylate in Pseudomonas.
    J Bacteriol. 1972 Nov;112(2):815-23 PMID: 4628746
  26. Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134.
    J Bacteriol. 1987 Jul;169(7):2950-5 PMID: 3036764
  27. Expression of naphthalene oxidation genes in Escherichia coli results in the biosynthesis of indigo.
    Science. 1983 Oct 14;222(4620):167-9 PMID: 6353574
  28. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5.
    J Bacteriol. 1978 Jul;135(1):227-38 PMID: 97269
  29. Cloning and characterization of a Pseudomonas mendocina KR1 gene cluster encoding toluene-4-monooxygenase.
    J Bacteriol. 1991 Sep;173(17):5315-27 PMID: 1885512
  30. Microbial transformation of 2,4,6-trinitrotoluene and other nitroaromatic compounds.
    Appl Environ Microbiol. 1976 Jun;31(6):949-58 PMID: 779650
  31. Degradation of 2,4-dinitrotoluene by the lignin-degrading fungus Phanerochaete chrysosporium.
    Appl Environ Microbiol. 1992 Jan;58(1):221-8 PMID: 1539977
  32. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
    Gene. 1985;33(1):103-19 PMID: 2985470
  33. Cloning of genes controlling alginate biosynthesis from a mucoid cystic fibrosis isolate of Pseudomonas aeruginosa.
    J Bacteriol. 1984 Jul;159(1):9-18 PMID: 6330052
  34. Toluene degradation by Pseudomonas putida F1. Nucleotide sequence of the todC1C2BADE genes and their expression in Escherichia coli.
    J Biol Chem. 1989 Sep 5;264(25):14940-6 PMID: 2670929
  35. Identification of nitroso compounds from biotransformation of 2,4-dinitrotoluene.
    Appl Environ Microbiol. 1984 Jun;47(6):1295-8 PMID: 16346568
  36. Isolation of TOL and RP4 recombinants by integrative suppression.
    J Bacteriol. 1978 Apr;134(1):270-7 PMID: 418059
  37. 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 PMID: 2981813
  38. Toluene degradation by Pseudomonas putida F1: genetic organization of the tod operon.
    Appl Environ Microbiol. 1988 Jun;54(6):1498-503 PMID: 2843094
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.6.1831-1837.1993
Published
1993-03-00
Pages
1831-7
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC203984
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.175.6.1831-1837.1993 PMC Full Text View journal details
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