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. 1989 Sep;171(9):5111–5116. doi: 10.1128/jb.171.9.5111-5116.1989

Degradation of phenol and m-toluate in Pseudomonas sp. strain EST1001 and its Pseudomonas putida transconjugants is determined by a multiplasmid system.

M A Kivisaar 1, J K Habicht 1, A L Heinaru 1
PMCID: PMC210324  PMID: 2768199

Abstract

The utilization of phenol, m-toluate, and salicylate (Phe+, mTol+, and Sal+ characters, respectively) in Pseudomonas sp. strain EST1001 is determined by the coordinated expression of genes placed in different plasmids, i.e., by a multiplasmid system. The natural multiplasmid strain EST1001 is phenotypically unstable. In its Phe-, mTol-, and Sal- segregants, the plasmid DNA underwent structural rearrangements without a marked loss of plasmid DNA, and the majority of segregants gave revertants. The genes specifying the degradation of phenol and m-toluate were transferable to P. putida PaW340, and in this strain a new multiplasmid system with definite structural changes was formed. The 17-kilobase transposable element, a part of the TOL plasmid pWWO present in the chromosome of PaW340, was inserted into the plasmid DNA in transconjugants. In addition, transconjugant EST1020 shared pWWO-like structures. Enzyme assays demonstrated that ortho-fission reactions were used by bacteria that grew on phenol, whereas m-toluate was catabolized by a meta-fission reaction. Salicylate was a functional inducer of the enzymes of both pathways. The expression of silent metabolic pathways of phenol or m-toluate degradation has been observed in EST1001 Phe- mTol+ and Phe+ mTol- transconjugants. The switchover of phenol degradation from the ortho- to the meta-pathway in EST1033 also showed the flexibility of genetic material in EST1001 transconjugants.

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Selected References

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

  1. Austen R. A., Dunn N. W. A comparative study of the NAH and TOL catabolic plasmids in Pseudomonas putida. Aust J Biol Sci. 1977 Aug;30(4):357–366. doi: 10.1071/bi9770357. [DOI] [PubMed] [Google Scholar]
  2. BAUCHOP T., ELSDEN S. R. The growth of micro-organisms in relation to their energy supply. J Gen Microbiol. 1960 Dec;23:457–469. doi: 10.1099/00221287-23-3-457. [DOI] [PubMed] [Google Scholar]
  3. Bagdasarian M., Timmis K. N. Host: vector systems for gene cloning in Pseudomonas. Curr Top Microbiol Immunol. 1982;96:47–67. doi: 10.1007/978-3-642-68315-2_4. [DOI] [PubMed] [Google Scholar]
  4. Bayley S. A., Duggleby C. J., Worsey M. J., Williams P. A., Hardy K. G., Broda P. Two modes of loss of the Tol function from Pseudomonas putida mt-2. Mol Gen Genet. 1977 Jul 20;154(2):203–204. doi: 10.1007/BF00330838. [DOI] [PubMed] [Google Scholar]
  5. Chakrabarty A. M., Friello D. A., Bopp L. H. Transposition of plasmid DNA segments specifying hydrocarbon degradation and their expression in various microorganisms. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3109–3112. doi: 10.1073/pnas.75.7.3109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chakrabarty A. M. Genetic basis of the biodegradation of salicylate in Pseudomonas. J Bacteriol. 1972 Nov;112(2):815–823. doi: 10.1128/jb.112.2.815-823.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chakrabarty A. M. Plasmids in Pseudomonas. Annu Rev Genet. 1976;10:7–30. doi: 10.1146/annurev.ge.10.120176.000255. [DOI] [PubMed] [Google Scholar]
  8. Chatterjee D. K., Chakrabarty A. M. Genetic rearrangements in plasmids specifying total degradation of chlorinated benzoic acids. Mol Gen Genet. 1982;188(2):279–285. doi: 10.1007/BF00332688. [DOI] [PubMed] [Google Scholar]
  9. Downing R., Broda P. A cleavage map of the TOL plasmid of Pseudomonas putida mt-2. Mol Gen Genet. 1979;177(1):189–191. doi: 10.1007/BF00267270. [DOI] [PubMed] [Google Scholar]
  10. Duggleby C. J., Bayley S. A., Worsey M. J., Williams P. A., Broda P. Molecular sizes and relationships of TOL plasmids in Pseudomonas. J Bacteriol. 1977 Jun;130(3):1274–1280. doi: 10.1128/jb.130.3.1274-1280.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dunn N. W., Gunsalus I. C. Transmissible plasmid coding early enzymes of naphthalene oxidation in Pseudomonas putida. J Bacteriol. 1973 Jun;114(3):974–979. doi: 10.1128/jb.114.3.974-979.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feist C. F., Hegeman G. D. Phenol and benzoate metabolism by Pseudomonas putida: regulation of tangential pathways. J Bacteriol. 1969 Nov;100(2):869–877. doi: 10.1128/jb.100.2.869-877.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Franklin F. C., Williams P. A. Construction of a partial diploid for the degradative pathway encoded by the TOL plasmid (pWWO) from Pseudomonas putida mt-2: evidence for the positive nature of the regulation by the xyIR gene. Mol Gen Genet. 1980 Jan;177(2):321–328. doi: 10.1007/BF00267445. [DOI] [PubMed] [Google Scholar]
  14. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hegeman G. D. Synthesis of the enzymes of the mandelate pathway by Pseudomonas putida. I. Synthesis of enzymes by the wild type. J Bacteriol. 1966 Mar;91(3):1140–1154. doi: 10.1128/jb.91.3.1140-1154.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hughes E. J., Bayly R. C., Skurray R. A. Characterization of a TOL-like plasmid from Alcaligenes eutrophus that controls expression of a chromosomally encoded p-cresol pathway. J Bacteriol. 1984 Apr;158(1):73–78. doi: 10.1128/jb.158.1.73-78.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jeenes D. J., Williams P. A. Excision and integration of degradative pathway genes from TOL plasmid pWW0. J Bacteriol. 1982 Apr;150(1):188–194. doi: 10.1128/jb.150.1.188-194.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Meulien P., Broda P. Identification of chromosomally integrated TOL DNA in cured derivatives of Pseudomonas putida PAW1. J Bacteriol. 1982 Nov;152(2):911–914. doi: 10.1128/jb.152.2.911-914.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meulien P., Downing R. G., Broda P. Excision of the 40kb segment of the TOL plasmid from Pseudomonas putida mt-2 involves direct repeats. Mol Gen Genet. 1981;184(1):97–101. doi: 10.1007/BF00271202. [DOI] [PubMed] [Google Scholar]
  22. Sala-Trepat J. M., Murray K., Williams P. A. The metabolic divergence in the meta cleavage of catechols by Pseudomonas putida NCIB 10015. Physiological significance and evolutionary implications. Eur J Biochem. 1972 Jul 24;28(3):347–356. doi: 10.1111/j.1432-1033.1972.tb01920.x. [DOI] [PubMed] [Google Scholar]
  23. Schwien U., Schmidt E. Improved degradation of monochlorophenols by a constructed strain. Appl Environ Microbiol. 1982 Jul;44(1):33–39. doi: 10.1128/aem.44.1.33-39.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tsuda M., Iino T. Genetic analysis of a transposon carrying toluene degrading genes on a TOL plasmid pWW0. Mol Gen Genet. 1987 Dec;210(2):270–276. doi: 10.1007/BF00325693. [DOI] [PubMed] [Google Scholar]
  25. Wheelis L. The genetics of dissimilarity pathways in Pseudomonas. Annu Rev Microbiol. 1975;29:505–524. doi: 10.1146/annurev.mi.29.100175.002445. [DOI] [PubMed] [Google Scholar]
  26. Williams P. A. Genetic interactions between mixed microbial populations. Philos Trans R Soc Lond B Biol Sci. 1982 Jun 11;297(1088):631–639. doi: 10.1098/rstb.1982.0066. [DOI] [PubMed] [Google Scholar]
  27. Williams P. A., Murray K. Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. J Bacteriol. 1974 Oct;120(1):416–423. doi: 10.1128/jb.120.1.416-423.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Williams P. A., Worsey M. J. Ubiquity of plasmids in coding for toluene and xylene metabolism in soil bacteria: evidence for the existence of new TOL plasmids. J Bacteriol. 1976 Mar;125(3):818–828. doi: 10.1128/jb.125.3.818-828.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wong C. L., Dunn N. W. Transmissible plasmid coding for the degradation of benzoate and m-toluate in Pseudomonas arvilla mt-2. Genet Res. 1974 Apr;23(2):227–232. doi: 10.1017/s0016672300014853. [DOI] [PubMed] [Google Scholar]
  30. Wong C. L., Leong R. W., Dunn N. W. Mutation to increased resistance to phenol in Pseudomonas putida. Biotechnol Bioeng. 1978 Jun;20(6):917–920. doi: 10.1002/bit.260200612. [DOI] [PubMed] [Google Scholar]
  31. Yen K. M., Gunsalus I. C. Plasmid gene organization: naphthalene/salicylate oxidation. Proc Natl Acad Sci U S A. 1982 Feb;79(3):874–878. doi: 10.1073/pnas.79.3.874. [DOI] [PMC free article] [PubMed] [Google Scholar]

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