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. 1993 Sep;59(9):2790–2794. doi: 10.1128/aem.59.9.2790-2794.1993

Variation in chlorobenzoate catabolism by Pseudomonas putida P111 as a consequence of genetic alterations.

V Brenner 1, B S Hernandez 1, D D Focht 1
PMCID: PMC182367  PMID: 8215353

Abstract

Pseudomonas putida P111 is able to utilize a broad range of monochlorinated, dichlorinated, and trichlorinated benzoates. The involvement of two separate dioxygenases was noted from data on plasmid profiles and DNA hybridization. The benzoate dioxygenase, which converts 3-chlorobenzoate (3-CB), 4-CB, and benzoate to the corresponding catechols via reduction of a dihydrodiol, was shown to be chromosomally coded. The chlorobenzoate-1,2-dioxygenase that converts ortho-chlorobenzoates to the corresponding catechols without the need of a functional dioldehydrogenase was shown to be encoded on plasmid pPB111 (75 kb). Cured strains were unable to utilize ortho-chlorobenzoates for growth. DNA hybridization data indicated that catabolism of the corresponding chlorocatechols was coded on chromosomal genes. Maintenance of plasmid pPB111 was dependent on the presence of ortho-chlorobenzoates in the growth media. A unique variant of P111 (P111D), able to grow on 3,5-dichlorobenzoate (3,5-DCB), was obtained by continuous subculturing from media containing progressively lower and higher concentrations of 3-CB and 3,5-DCB, respectively. The low frequency of segregants able to grow on 2,5-DCB, 2,3-DCB, and 2,3, 5-trichlorobenzoate was evident by lag periods greater than 200 h. Continued subculture on 3,5-DCB resulted in the formation of new plasmid pPH111 (120 kb), which was homologous to pPB111. A probe from the clc operon, which encodes for the chlorocatechol pathway, hybridized to plasmid pPH111 and to the chromosome of the wild-type strain P111 but not to its plasmid pPB111 nor to the chromosome of strain P111A, which had lost the ability to utilize chlorobenzoates.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Adams R. H., Huang C. M., Higson F. K., Brenner V., Focht D. D. Construction of a 3-chlorobiphenyl-utilizing recombinant from an intergeneric mating. Appl Environ Microbiol. 1992 Feb;58(2):647–654. doi: 10.1128/aem.58.2.647-654.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adriaens P., Kohler H. P., Kohler-Staub D., Focht D. D. Bacterial dehalogenation of chlorobenzoates and coculture biodegradation of 4,4'-dichlorobiphenyl. Appl Environ Microbiol. 1989 Apr;55(4):887–892. doi: 10.1128/aem.55.4.887-892.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chatterjee D. K., Chakrabarty A. M. Restriction mapping of a chlorobenzoate degradative plasmid and molecular cloning of the degradative genes. Gene. 1984 Feb;27(2):173–181. doi: 10.1016/0378-1119(84)90138-0. [DOI] [PubMed] [Google Scholar]
  5. Chatterjee D. K., Kellogg S. T., Hamada S., Chakrabarty A. M. Plasmid specifying total degradation of 3-chlorobenzoate by a modified ortho pathway. J Bacteriol. 1981 May;146(2):639–646. doi: 10.1128/jb.146.2.639-646.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Dorn E., Hellwig M., Reineke W., Knackmuss H. J. Isolation and characterization of a 3-chlorobenzoate degrading pseudomonad. Arch Microbiol. 1974;99(1):61–70. doi: 10.1007/BF00696222. [DOI] [PubMed] [Google Scholar]
  8. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  9. Engesser K. H., Schulte P. Degradation of 2-bromo-, 2-chloro- and 2-fluorobenzoate by Pseudomonas putida CLB 250. FEMS Microbiol Lett. 1989 Jul 15;51(1):143–147. doi: 10.1016/0378-1097(89)90497-7. [DOI] [PubMed] [Google Scholar]
  10. Fetzner S., Müller R., Lingens F. Degradation of 2-chlorobenzoate by Pseudomonas cepacia 2CBS. Biol Chem Hoppe Seyler. 1989 Nov;370(11):1173–1182. doi: 10.1515/bchm3.1989.370.2.1173. [DOI] [PubMed] [Google Scholar]
  11. Fetzner S., Müller R., Lingens F. Purification and some properties of 2-halobenzoate 1,2-dioxygenase, a two-component enzyme system from Pseudomonas cepacia 2CBS. J Bacteriol. 1992 Jan;174(1):279–290. doi: 10.1128/jb.174.1.279-290.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Focht D. D., Shelton D. Growth kinetics of Pseudomonas alcaligenes C-0 relative to inoculation and 3-chlorobenzoate metabolism in soil. Appl Environ Microbiol. 1987 Aug;53(8):1846–1849. doi: 10.1128/aem.53.8.1846-1849.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frantz B., Chakrabarty A. M. Organization and nucleotide sequence determination of a gene cluster involved in 3-chlorocatechol degradation. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4460–4464. doi: 10.1073/pnas.84.13.4460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hartmann J., Reineke W., Knackmuss H. J. Metabolism of 3-chloro-, 4-chloro-, and 3,5-dichlorobenzoate by a pseudomonad. Appl Environ Microbiol. 1979 Mar;37(3):421–428. doi: 10.1128/aem.37.3.421-428.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hernandez B. S., Higson F. K., Kondrat R., Focht D. D. Metabolism of and inhibition by chlorobenzoates in Pseudomonas putida P111. Appl Environ Microbiol. 1991 Nov;57(11):3361–3366. doi: 10.1128/aem.57.11.3361-3366.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hickey W. J., Focht D. D. Degradation of mono-, di-, and trihalogenated benzoic acids by Pseudomonas aeruginosa JB2. Appl Environ Microbiol. 1990 Dec;56(12):3842–3850. doi: 10.1128/aem.56.12.3842-3850.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Marks T. S., Smith A. R., Quirk A. V. Degradation of 4-Chlorobenzoic Acid by Arthrobacter sp. Appl Environ Microbiol. 1984 Nov;48(5):1020–1025. doi: 10.1128/aem.48.5.1020-1025.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nordström K., Molin S., Aagaard-Hansen H. Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid. 1980 Sep;4(2):215–227. doi: 10.1016/0147-619x(80)90011-6. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Pertsova R. N., Kunc F., Golovleva L. A. Degradation of 3-chlorobenzoate in soil by pseudomonads carrying biodegradative plasmids. Folia Microbiol (Praha) 1984;29(3):242–247. doi: 10.1007/BF02877315. [DOI] [PubMed] [Google Scholar]
  21. Reineke W., Knackmuss H. J. Microbial degradation of haloaromatics. Annu Rev Microbiol. 1988;42:263–287. doi: 10.1146/annurev.mi.42.100188.001403. [DOI] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Sylvestre M., Mailhiot K., Ahmad D., Massé R. Isolation and preliminary characterization of a 2-chlorobenzoate degrading Pseudomonas. Can J Microbiol. 1989 Apr;35(4):439–443. doi: 10.1139/m89-067. [DOI] [PubMed] [Google Scholar]
  24. Wyndham R. C., Straus N. A. Chlorobenzoate catabolism and interactions between Alcaligenes and Pseudomonas species from Bloody Run Creek. Arch Microbiol. 1988;150(3):230–236. doi: 10.1007/BF00407785. [DOI] [PubMed] [Google Scholar]
  25. van den Tweel W. J., Kok J. B., de Bont J. A. Reductive dechlorination of 2,4-dichlorobenzoate to 4-chlorobenzoate and hydrolytic dehalogenation of 4-chloro-, 4-bromo-, and 4-iodobenzoate by Alcaligenes denitrificans NTB-1. Appl Environ Microbiol. 1987 Apr;53(4):810–815. doi: 10.1128/aem.53.4.810-815.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. van der Meer J. R., van Neerven A. R., de Vries E. J., de Vos W. M., Zehnder A. J. 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. doi: 10.1128/jb.173.1.6-15.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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