Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Nov;62(11):4073–4080. doi: 10.1128/aem.62.11.4073-4080.1996

Regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene by naphthalene dioxygenase from Pseudomonas sp. strain NCIB 9816-4.

S M Resnick 1, D T Gibson 1
PMCID: PMC168229  PMID: 8899998

Abstract

The regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene was examined with mutant and recombinant strains expressing naphthalene dioxygenase from Pseudomonas sp. strain NCIB 9816-4. The initial oxidation products were isolated and identified by gas chromatography-mass spectrometry and nuclear magnetic resonance spectrometry. Salicylate-induced cells of Pseudomonas sp. strain 9816/11 and isopropyl-beta-D-thiogalactopyranoside-induced cells of Escherichia coli JM109(DE3)(pDTG141) oxidized fluorene to (+)-(3S,4R)-cis-3,4-dihydroxy-3,4-dihydrofluorene (80 to 90% relative yield; > 95% enantiomeric excess [ee]) and 9-fluorenol (< 10% yield). The same cells oxidized dibenzofuran to (1R,2S)-cis-1,2-dihydroxy-1, 2-dihydrodibenzofuran (60 to 70% yield; > 95% ee) and (3S,4R)-cis-3, 4-dihydroxy-3,4-dihydrodibenzofuran (30 to 40% yield; > 95% ee). Induced cells of both strains, as well as the purified dioxygenase, also oxidized dibenzothiophene to (+)-(1R,2S)-cis-1,2-dihydroxy-1, 2-dihydrodibenzothiophene (84 to 87% yield; > 95% ee) and dibenzothiophene sulfoxide (< 15% yield). The major reaction catalyzed by naphthalene dioxygenase with each substrate was stereospecific dihydroxylation in which the cis-dihydrodiols were of identical regiochemistry and of R configuration at the benzylic center adjacent to the bridgehead carbon atom. The regiospecific oxidation of dibenzofuran differed from that of the other substrates in that a significant amount of the minor cis-3,4-dihydrodiol regioisomer was formed. The results indicate that although the absolute stereochemistry of the cis-diene diols was the same, the nature of the bridging atom or heteroatom influenced the regiospecificity of the reactions catalyzed by naphthalene dioxygenase.

Full Text

The Full Text of this article is available as a PDF (243.3 KB).

Selected References

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

  1. Akhtar M. N., Boyd D. R., Thompson N. J., Koreeda M., Gibson D. T., Mahadevan V., Jerina D. M. Absolute sterochemistry of the dihydroanthracene-cis- and -trans-1,2-diols produced from anthracene by mammals and bacteria. J Chem Soc Perkin 1. 1975;(23):2506–2511. [PubMed] [Google Scholar]
  2. Cerniglia C. E., Morgan J. C., Gibson D. T. Bacterial and fungal oxidation of dibenzofuran. Biochem J. 1979 Apr 15;180(1):175–185. doi: 10.1042/bj1800175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chapman P. J., Shelton M., Grifoll M., Selifonov S. Fossil fuel biodegradation: laboratory studies. Environ Health Perspect. 1995 Jun;103 (Suppl 5):79–83. doi: 10.1289/ehp.95103s479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Denome S. A., Stanley D. C., Olson E. S., Young K. D. Metabolism of dibenzothiophene and naphthalene in Pseudomonas strains: complete DNA sequence of an upper naphthalene catabolic pathway. J Bacteriol. 1993 Nov;175(21):6890–6901. doi: 10.1128/jb.175.21.6890-6901.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eaton R. W., Chapman P. J. Bacterial metabolism of naphthalene: construction and use of recombinant bacteria to study ring cleavage of 1,2-dihydroxynaphthalene and subsequent reactions. J Bacteriol. 1992 Dec;174(23):7542–7554. doi: 10.1128/jb.174.23.7542-7554.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eaton R. W., Selifonov S. A. Biotransformation of 6,6-Dimethylfulvene by Pseudomonas putida RE213. Appl Environ Microbiol. 1996 Mar;62(3):756–760. doi: 10.1128/aem.62.3.756-760.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Engesser K. H., Strubel V., Christoglou K., Fischer P., Rast H. G. Dioxygenolytic cleavage of aryl ether bonds: 1,10-dihydro-1,10-dihydroxyfluoren-9-one, a novel arene dihydrodiol as evidence for angular dioxygenation of dibenzofuran. FEMS Microbiol Lett. 1989 Nov;53(1-2):205–209. doi: 10.1016/0378-1097(89)90392-3. [DOI] [PubMed] [Google Scholar]
  8. Erickson B. D., Mondello F. J. Enhanced biodegradation of polychlorinated biphenyls after site-directed mutagenesis of a biphenyl dioxygenase gene. Appl Environ Microbiol. 1993 Nov;59(11):3858–3862. doi: 10.1128/aem.59.11.3858-3862.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Erickson B. D., Mondello F. J. Nucleotide sequencing and transcriptional mapping of the genes encoding biphenyl dioxygenase, a multicomponent polychlorinated-biphenyl-degrading enzyme in Pseudomonas strain LB400. J Bacteriol. 1992 May;174(9):2903–2912. doi: 10.1128/jb.174.9.2903-2912.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fortnagel P., Harms H., Wittich R. M., Krohn S., Meyer H., Sinnwell V., Wilkes H., Francke W. Metabolism of Dibenzofuran by Pseudomonas sp. Strain HH69 and the Mixed Culture HH27. Appl Environ Microbiol. 1990 Apr;56(4):1148–1156. doi: 10.1128/aem.56.4.1148-1156.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gibson D. T., Cruden D. L., Haddock J. D., Zylstra G. J., Brand J. M. Oxidation of polychlorinated biphenyls by Pseudomonas sp. strain LB400 and Pseudomonas pseudoalcaligenes KF707. J Bacteriol. 1993 Jul;175(14):4561–4564. doi: 10.1128/jb.175.14.4561-4564.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gibson D. T., Resnick S. M., Lee K., Brand J. M., Torok D. S., Wackett L. P., Schocken M. J., Haigler B. E. Desaturation, dioxygenation, and monooxygenation reactions catalyzed by naphthalene dioxygenase from Pseudomonas sp. strain 9816-4. J Bacteriol. 1995 May;177(10):2615–2621. doi: 10.1128/jb.177.10.2615-2621.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grifoll M., Casellas M., Bayona J. M., Solanas A. M. Isolation and characterization of a fluorene-degrading bacterium: identification of ring oxidation and ring fission products. Appl Environ Microbiol. 1992 Sep;58(9):2910–2917. doi: 10.1128/aem.58.9.2910-2917.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Grifoll M., Selifonov S. A., Gatlin C. V., Chapman P. J. Actions of a versatile fluorene-degrading bacterial isolate on polycyclic aromatic compounds. Appl Environ Microbiol. 1995 Oct;61(10):3711–3723. doi: 10.1128/aem.61.10.3711-3723.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Haddock J. D., Horton J. R., Gibson D. T. Dihydroxylation and dechlorination of chlorinated biphenyls by purified biphenyl 2,3-dioxygenase from Pseudomonas sp. strain LB400. J Bacteriol. 1995 Jan;177(1):20–26. doi: 10.1128/jb.177.1.20-26.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jeffrey A. M., Yeh H. J., Jerina D. M., Patel T. R., Davey J. F., Gibson D. T. Initial reactions in the oxidation of naphthalene by Pseudomonas putida. Biochemistry. 1975 Feb 11;14(3):575–584. doi: 10.1021/bi00674a018. [DOI] [PubMed] [Google Scholar]
  17. Jerina D. M., Daly J. W., Jeffrey A. M., Gibson D. T. Cis-1,2-dihydroxy-1,2-dihydronaphthalene: a bacterial metabolite from naphthalene. Arch Biochem Biophys. 1971 Jan;142(1):394–396. doi: 10.1016/0003-9861(71)90298-0. [DOI] [PubMed] [Google Scholar]
  18. Jerina D. M., Selander H., Yagi H., Wells M. C., Davey J. F., Mahadevan V., Gibson D. T. Dihydrodiols from anthracene and phenanthrene. J Am Chem Soc. 1976 Sep 15;98(19):5988–5996. doi: 10.1021/ja00435a035. [DOI] [PubMed] [Google Scholar]
  19. Khan A. A., Wang R. F., Cao W. W., Franklin W., Cerniglia C. E. Reclassification of a polycyclic aromatic hydrocarbon-metabolizing bacterium, Beijerinckia sp. strain B1, as Sphingomonas yanoikuyae by fatty acid analysis, protein pattern analysis, DNA-DNA hybridization, and 16S ribosomal DNA sequencing. Int J Syst Bacteriol. 1996 Apr;46(2):466–469. doi: 10.1099/00207713-46-2-466. [DOI] [PubMed] [Google Scholar]
  20. Klecka G. M., Gibson D. T. Metabolism of dibenzo[1,4]dioxan by a Pseudomonas species. Biochem J. 1979 Jun 15;180(3):639–645. doi: 10.1042/bj1800639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kurkela S., Lehväslaiho H., Palva E. T., Teeri T. H. Cloning, nucleotide sequence and characterization of genes encoding naphthalene dioxygenase of Pseudomonas putida strain NCIB9816. Gene. 1988 Dec 20;73(2):355–362. doi: 10.1016/0378-1119(88)90500-8. [DOI] [PubMed] [Google Scholar]
  22. Laborde A. L., Gibson D. T. Metabolism of dibenzothiophene by a Beijerinckia species. Appl Environ Microbiol. 1977 Dec;34(6):783–790. doi: 10.1128/aem.34.6.783-790.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee K., Brand J. M., Gibson D. T. Stereospecific sulfoxidation by toluene and naphthalene dioxygenases. Biochem Biophys Res Commun. 1995 Jul 6;212(1):9–15. doi: 10.1006/bbrc.1995.1928. [DOI] [PubMed] [Google Scholar]
  24. Lee K., Gibson D. T. Toluene and ethylbenzene oxidation by purified naphthalene dioxygenase from Pseudomonas sp. strain NCIB 9816-4. Appl Environ Microbiol. 1996 Sep;62(9):3101–3106. doi: 10.1128/aem.62.9.3101-3106.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mahaffey W. R., Gibson D. T., Cerniglia C. E. Bacterial oxidation of chemical carcinogens: formation of polycyclic aromatic acids from benz[a]anthracene. Appl Environ Microbiol. 1988 Oct;54(10):2415–2423. doi: 10.1128/aem.54.10.2415-2423.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Monticello D. J., Bakker D., Finnerty W. R. Plasmid-mediated degradation of dibenzothiophene by Pseudomonas species. Appl Environ Microbiol. 1985 Apr;49(4):756–760. doi: 10.1128/aem.49.4.756-760.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mueller J. G., Middaugh D. P., Lantz S. E., Chapman P. J. Biodegradation of creosote and pentachlorophenol in contaminated groundwater: chemical and biological assessment. Appl Environ Microbiol. 1991 May;57(5):1277–1285. doi: 10.1128/aem.57.5.1277-1285.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pothuluri J. V., Freeman J. P., Evans F. E., Cerniglia C. E. Biotransformation of fluorene by the fungus Cunninghamella elegans. Appl Environ Microbiol. 1993 Jun;59(6):1977–1980. doi: 10.1128/aem.59.6.1977-1980.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Resnick S. M., Gibson D. T. Oxidation of 6,7-dihydro-5H-benzocycloheptene by bacterial strains expressing naphthalene dioxygenase, biphenyl dioxygenase, and toluene dioxygenase yields homochiral monol or cis-diol enantiomers as major products. Appl Environ Microbiol. 1996 Apr;62(4):1364–1368. doi: 10.1128/aem.62.4.1364-1368.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Resnick S. M., Gibson D. T. Regio- and stereospecific oxidation of 9,10-dihydroanthracene and 9,10-dihydrophenanthrene by naphthalene dioxygenase: structure and absolute stereochemistry of metabolites. Appl Environ Microbiol. 1996 Sep;62(9):3355–3359. doi: 10.1128/aem.62.9.3355-3359.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Resnick S. M., Torok D. S., Gibson D. T. Oxidation of carbazole to 3-hydroxycarbazole by naphthalene 1,2-dioxygenase and biphenyl 2,3-dioxygenase. FEMS Microbiol Lett. 1993 Nov 1;113(3):297–302. doi: 10.1111/j.1574-6968.1993.tb06530.x. [DOI] [PubMed] [Google Scholar]
  32. Resnick S. M., Torok D. S., Lee K., Brand J. M., Gibson D. T. Regiospecific and stereoselective hydroxylation of 1-indanone and 2-indanone by naphthalene dioxygenase and toluene dioxygenase. Appl Environ Microbiol. 1994 Sep;60(9):3323–3328. doi: 10.1128/aem.60.9.3323-3328.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Selifonov S. A., Grifoll M., Eaton R. W., Chapman P. J. Oxidation of naphthenoaromatic and methyl-substituted aromatic compounds by naphthalene 1,2-dioxygenase. Appl Environ Microbiol. 1996 Feb;62(2):507–514. doi: 10.1128/aem.62.2.507-514.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Selifonov S. A., Grifoll M., Gurst J. E., Chapman P. J. Isolation and characterization of (+)-1,1a-dihydroxy-1-hydrofluoren-9-one formed by angular dioxygenation in the bacterial catabolism of fluorene. Biochem Biophys Res Commun. 1993 May 28;193(1):67–76. doi: 10.1006/bbrc.1993.1591. [DOI] [PubMed] [Google Scholar]
  35. Simon M. J., Osslund T. D., Saunders R., Ensley B. D., Suggs S., Harcourt A., Suen W. C., Cruden D. L., Gibson D. T., Zylstra G. J. Sequences of genes encoding naphthalene dioxygenase in Pseudomonas putida strains G7 and NCIB 9816-4. Gene. 1993 May 15;127(1):31–37. doi: 10.1016/0378-1119(93)90613-8. [DOI] [PubMed] [Google Scholar]
  36. Simoni S., Klinke S., Zipper C., Angst W., Kohler H. E. Enantioselective Metabolism of Chiral 3-Phenylbutyric Acid, an Intermediate of Linear Alkylbenzene Degradation, by Rhodococcus rhodochrous PB1. Appl Environ Microbiol. 1996 Mar;62(3):749–755. doi: 10.1128/aem.62.3.749-755.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Strubel V., Engesser K. H., Fischer P., Knackmuss H. J. 3-(2-hydroxyphenyl)catechol as substrate for proximal meta ring cleavage in dibenzofuran degradation by Brevibacterium sp. strain DPO 1361. J Bacteriol. 1991 Mar;173(6):1932–1937. doi: 10.1128/jb.173.6.1932-1937.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Taira K., Hirose J., Hayashida S., Furukawa K. Analysis of bph operon from the polychlorinated biphenyl-degrading strain of Pseudomonas pseudoalcaligenes KF707. J Biol Chem. 1992 Mar 5;267(7):4844–4853. [PubMed] [Google Scholar]
  39. Torok D. S., Resnick S. M., Brand J. M., Cruden D. L., Gibson D. T. Desaturation and oxygenation of 1,2-dihydronaphthalene by toluene and naphthalene dioxygenase. J Bacteriol. 1995 Oct;177(20):5799–5805. doi: 10.1128/jb.177.20.5799-5805.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wackett L. P., Kwart L. D., Gibson D. T. Benzylic monooxygenation catalyzed by toluene dioxygenase from Pseudomonas putida. Biochemistry. 1988 Feb 23;27(4):1360–1367. doi: 10.1021/bi00404a041. [DOI] [PubMed] [Google Scholar]
  41. Whited G. M., Downie J. C., Hudlicky T., Fearnley S. P., Dudding T. C., Olivo H. F., Parker D. Oxidation of 2-methoxynaphthalene by toluene, naphthalene and biphenyl dioxygenases:structure and absolute stereochemistry of metabolites. Bioorg Med Chem. 1994 Jul;2(7):727–734. doi: 10.1016/0968-0896(94)85024-0. [DOI] [PubMed] [Google Scholar]
  42. Yen K. M., Serdar C. M. Genetics of naphthalene catabolism in pseudomonads. Crit Rev Microbiol. 1988;15(3):247–268. doi: 10.3109/10408418809104459. [DOI] [PubMed] [Google Scholar]

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

RESOURCES