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. 1980 Aug;143(2):816–824. doi: 10.1128/jb.143.2.816-824.1980

p-Cymene pathway in Pseudomonas putida: selective enrichment of defective mutants by using halogenated substrate analogs.

G J Wigmore, D W Ribbons
PMCID: PMC294369  PMID: 7204334

Abstract

Several classes of mutants of Pseudomonas putida (JT810) defective in the utilization of p-cymene as sole carbon source have been isolated. Selective enrichment of the mutants and for strains putatively cured of a degradative plasmid was achieved by incubation of cells in minimal growth media containing p-cymene (or p-cumate) and various halogenated analogs of the growth substrates or pathway intermediates. Analogs which led to successful enrichments included: p-chlorotoluene, p-bromotoluene, alpha-chloro-p-xylene, and p-iodobenzoate. A mutant strain, PpJT811, constitutive for the p-cymene pathway gave significantly greater enrichments of defective mutants than the wild-type parent PpJT810 after incubation with the halogenated analogs. It is suggested that the defective mutants are enriched because of the genetic alterations they possess, which confer immunity to a lethal synthesis performed by transformation of the analogs in clones possessing an intact p-cymene pathway. A nomenclature for the genetic organization of p-cymene pathway is described.

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Selected References

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

  1. Apirion D. The two-way selection of mutants and revertants in respect of acetate utilization and resistance to fluoro-acetate in Aspergillus nidulans. Genet Res. 1965 Nov;6(3):317–329. doi: 10.1017/s0016672300004213. [DOI] [PubMed] [Google Scholar]
  2. Armitt S., McCullough W., Roberts C. F. Analysis of acetate non-utilizing (acu) mutants in Aspergillus nidulans. J Gen Microbiol. 1976 Feb;92(2):263–282. doi: 10.1099/00221287-92-2-263. [DOI] [PubMed] [Google Scholar]
  3. Bayly R. C., Wigmore G. J. Metabolism of phenol and cresols by mutants of Pseudomonas putida. J Bacteriol. 1973 Mar;113(3):1112–1120. doi: 10.1128/jb.113.3.1112-1120.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carhart G., Hegeman G. Improved method of selection for mutants of Pseudomonas putida. Appl Microbiol. 1975 Dec;30(6):1046–1047. doi: 10.1128/am.30.6.1046-1047.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chapman P. J., Ribbons D. W. Metabolism of resorcinylic compounds by bacteria: orcinol pathway in Pseudomonas putida. J Bacteriol. 1976 Mar;125(3):975–984. doi: 10.1128/jb.125.3.975-984.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collinsworth W. L., Chapman P. J., Dagley S. Stereospecific enzymes in the degradation of aromatic compounds by pseudomonas putida. J Bacteriol. 1973 Feb;113(2):922–931. doi: 10.1128/jb.113.2.922-931.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DeFrank J. J., Ribbons D. W. p-Cymene pathway in Pseudomonas putida: ring cleavage of 2,3-dihydroxy-p-cumate and subsequent reactions. J Bacteriol. 1977 Mar;129(3):1365–1374. doi: 10.1128/jb.129.3.1365-1374.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeFrank J. J., Ribbons D. W. p-cymene pathway in Pseudomonas putida: initial reactions. J Bacteriol. 1977 Mar;129(3):1356–1364. doi: 10.1128/jb.129.3.1356-1364.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Demerec M., Adelberg E. A., Clark A. J., Hartman P. E. A proposal for a uniform nomenclature in bacterial genetics. Genetics. 1966 Jul;54(1):61–76. doi: 10.1093/genetics/54.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. FUKASAWA T., NIKAIDO H. Galactose-sensitive mutants of Salmonella. II. Bacteriolysis induced by galactose. Biochim Biophys Acta. 1961 Apr 15;48:470–483. doi: 10.1016/0006-3002(61)90045-2. [DOI] [PubMed] [Google Scholar]
  11. Freedberg W. B., Kistler W. S., Lin E. C. Lethal synthesis of methylglyoxal by Escherichia coli during unregulated glycerol metabolism. J Bacteriol. 1971 Oct;108(1):137–144. doi: 10.1128/jb.108.1.137-144.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gilbert W., Müller-Hill B. Isolation of the lac repressor. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1891–1898. doi: 10.1073/pnas.56.6.1891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hynes M. J., Pateman J. A. The genetic analysis of regulation of amidase synthesis in Aspergillus nidulans. II. Mutants resistant to fluoroacetamide. Mol Gen Genet. 1970;108(2):107–116. doi: 10.1007/BF02430517. [DOI] [PubMed] [Google Scholar]
  14. Jones-Mortimer M. C., Kornberg H. L. Order of genes adjacent to ptsX on the E. coli genome. Proc R Soc Lond B Biol Sci. 1976 May 18;193(1112):313–315. doi: 10.1098/rspb.1976.0049. [DOI] [PubMed] [Google Scholar]
  15. KING E. O., WARD M. K., RANEY D. E. Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med. 1954 Aug;44(2):301–307. [PubMed] [Google Scholar]
  16. LUBIN M. Enrichment of auxotrophic mutant populations by recycling. J Bacteriol. 1962 Mar;83:696–697. doi: 10.1128/jb.83.3.696-697.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Malamy M. H. Frameshift mutations in the lactose operon of E. coli. Cold Spring Harb Symp Quant Biol. 1966;31:189–201. doi: 10.1101/sqb.1966.031.01.027. [DOI] [PubMed] [Google Scholar]
  18. NIKAIDO H. Galactose-sensitive mutants of Salmonella. I. Metabolism of galactose. Biochim Biophys Acta. 1961 Apr 15;48:460–469. doi: 10.1016/0006-3002(61)90044-0. [DOI] [PubMed] [Google Scholar]
  19. Ornston L. N., Ornston M. K., Chou G. Isolation of spontaneous mutant strains of Pseudomonas putida. Biochem Biophys Res Commun. 1969 Jul 7;36(1):179–184. doi: 10.1016/0006-291x(69)90666-4. [DOI] [PubMed] [Google Scholar]
  20. Smith T. F., Sadler J. R. The nature of lactose operator constitive mutations. J Mol Biol. 1971 Jul 28;59(2):273–305. doi: 10.1016/0022-2836(71)90051-9. [DOI] [PubMed] [Google Scholar]
  21. Stanier R. Y., Palleroni N. J., Doudoroff M. The aerobic pseudomonads: a taxonomic study. J Gen Microbiol. 1966 May;43(2):159–271. doi: 10.1099/00221287-43-2-159. [DOI] [PubMed] [Google Scholar]

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