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Applied and Environmental Microbiology, April 2004, p. 2391-2397, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2391-2397.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genome Shuffling Improves Degradation of the Anthropogenic Pesticide Pentachlorophenol by Sphingobium chlorophenolicum ATCC 39723

MingHua Dai and Shelley D. Copley*

Department of Molecular, Cellular, and Developmental Biology and Cooperative Institute for Research in Environmental Sciences, University of Colorado at Boulder, Boulder, Colorado 80309

Received 8 December 2003/ Accepted 16 December 2003

Pentachlorophenol (PCP), a highly toxic anthropogenic pesticide, can be mineralized by Sphingobium chlorophenolicum, a gram-negative bacterium isolated from PCP-contaminated soil. However, degradation of PCP is slow and S. chlorophenolicum cannot tolerate high levels of PCP. We have used genome shuffling to improve the degradation of PCP by S. chlorophenolicum. We have obtained several strains that degrade PCP faster and tolerate higher levels of PCP than the wild-type strain. Several strains obtained after the third round of shuffling can grow on one-quarter-strength tryptic soy broth plates containing 6 to 8 mM PCP, while the original strain cannot grow in the presence of PCP at concentrations higher than 0.6 mM. Some of the mutants are able to completely degrade 3 mM PCP in one-quarter-strength tryptic soy broth, whereas no degradation can be achieved by the wild-type strain. Analysis of several improved strains suggests that the improved phenotypes are due to various combinations of mutations leading to an enhanced growth rate, constitutive expression of the PCP degradation genes, and enhanced resistance to the toxicity of PCP and its metabolites.

* Corresponding author. Mailing address: Cooperative Institute for Research in Environmental Sciences, Campus Box 216, University of Colorado at Boulder, Boulder, CO 80309-0216. Phone: (303) 492-6328. Fax: (303) 492-1149. E-mail: shelley{at}cires.colorado.edu.

Applied and Environmental Microbiology, April 2004, p. 2391-2397, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2391-2397.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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