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. 1994 Nov;60(11):4148–4154. doi: 10.1128/aem.60.11.4148-4154.1994

Metabolism of chlorofluorocarbons and polybrominated compounds by Pseudomonas putida G786(pHG-2) via an engineered metabolic pathway.

H G Hur 1, M J Sadowsky 1, L P Wackett 1
PMCID: PMC201949  PMID: 7993096

Abstract

The recombinant bacterium Pseudomonas putida G786(pHG-2) metabolizes pentachloroethane to glyoxylate and carbon dioxide, using cytochrome P-450CAM and toluene dioxygenase to catalyze consecutive reductive and oxidative dehalogenation reactions (L.P. Wackett, M.J. Sadowsky, L.N. Newman, H.-G. Hur, and S. Li, Nature [London] 368:627-629, 1994). The present study investigated metabolism of brominated and chlorofluorocarbon compounds by the recombinant strain. Under anaerobic conditions, P. putida G786(pHG-2) reduced 1,1,2,2-tetrabromoethane, 1,2-dibromo-1,2-dichloroethane, and 1,1,1,2-tetrachloro-2,2-difluoroethane to products bearing fewer halogen substituents. Under aerobic conditions, P. putida G786(pHG-2) oxidized cis- and trans-1,2-dibromoethenes, 1,1-dichloro-2,2-difluoroethene, and 1,2-dichloro-1-fluoroethene. Several compounds were metabolized by sequential reductive and oxidative reactions via the constructed metabolic pathway. For example, 1,1,2,2-tetrabromoethane was reduced by cytochrome P-450CAM to 1,2-dibromoethenes, which were subsequently oxidized by toluene dioxygenase. The same pathway metabolized 1,1,1,2-tetrachloro-2,2-difluoroethane to oxalic acid as one of the final products. The results obtained in this study indicate that P. putida G786(pHG-2) metabolizes polyfluorinated, chlorinated, and brominated compounds and further demonstrates the value of using a knowledge of catabolic enzymes and recombinant DNA technology to construct useful metabolic pathways.

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Selected References

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  1. Arciero D., Vannelli T., Logan M., Hooper A. B. Degradation of trichloroethylene by the ammonia-oxidizing bacterium Nitrosomonas europaea. Biochem Biophys Res Commun. 1989 Mar 15;159(2):640–643. doi: 10.1016/0006-291x(89)90042-9. [DOI] [PubMed] [Google Scholar]
  2. Castro C. E., Wade R. S., Belser N. O. Biodehalogenation: reactions of cytochrome P-450 with polyhalomethanes. Biochemistry. 1985 Jan 1;24(1):204–210. doi: 10.1021/bi00322a029. [DOI] [PubMed] [Google Scholar]
  3. Fathepure B. Z., Vogel T. M. Complete degradation of polychlorinated hydrocarbons by a two-stage biofilm reactor. Appl Environ Microbiol. 1991 Dec;57(12):3418–3422. doi: 10.1128/aem.57.12.3418-3422.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fox B. G., Borneman J. G., Wackett L. P., Lipscomb J. D. Haloalkene oxidation by the soluble methane monooxygenase from Methylosinus trichosporium OB3b: mechanistic and environmental implications. Biochemistry. 1990 Jul 10;29(27):6419–6427. doi: 10.1021/bi00479a013. [DOI] [PubMed] [Google Scholar]
  5. Lefever M. R., Wackett L. P. Oxidation of low molecular weight chloroalkanes by cytochrome P450CAM. Biochem Biophys Res Commun. 1994 May 30;201(1):373–378. doi: 10.1006/bbrc.1994.1711. [DOI] [PubMed] [Google Scholar]
  6. Li S., Wackett L. P. Reductive dehalogenation by cytochrome P450CAM: substrate binding and catalysis. Biochemistry. 1993 Sep 14;32(36):9355–9361. doi: 10.1021/bi00087a014. [DOI] [PubMed] [Google Scholar]
  7. Li S., Wackett L. P. Trichloroethylene oxidation by toluene dioxygenase. Biochem Biophys Res Commun. 1992 May 29;185(1):443–451. doi: 10.1016/s0006-291x(05)81005-8. [DOI] [PubMed] [Google Scholar]
  8. Orser C. S., Dutton J., Lange C., Jablonski P., Xun L., Hargis M. Characterization of a Flavobacterium glutathione S-transferase gene involved reductive dechlorination. J Bacteriol. 1993 May;175(9):2640–2644. doi: 10.1128/jb.175.9.2640-2644.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Orser C. S., Lange C. C., Xun L., Zahrt T. C., Schneider B. J. Cloning, sequence analysis, and expression of the Flavobacterium pentachlorophenol-4-monooxygenase gene in Escherichia coli. J Bacteriol. 1993 Jan;175(2):411–416. doi: 10.1128/jb.175.2.411-416.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Vannelli T., Logan M., Arciero D. M., Hooper A. B. Degradation of halogenated aliphatic compounds by the ammonia- oxidizing bacterium Nitrosomonas europaea. Appl Environ Microbiol. 1990 Apr;56(4):1169–1171. doi: 10.1128/aem.56.4.1169-1171.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wackett L. P., Gibson D. T. Degradation of trichloroethylene by toluene dioxygenase in whole-cell studies with Pseudomonas putida F1. Appl Environ Microbiol. 1988 Jul;54(7):1703–1708. doi: 10.1128/aem.54.7.1703-1708.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wackett L. P., Sadowsky M. J., Newman L. M., Hur H. G., Li S. Metabolism of polyhalogenated compounds by a genetically engineered bacterium. Nature. 1994 Apr 14;368(6472):627–629. doi: 10.1038/368627a0. [DOI] [PubMed] [Google Scholar]

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