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Applied and Environmental Microbiology, May 1999, p. 2163-2169, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Construction and Characterization of Two Recombinant Bacteria That Grow on ortho- and para-Substituted Chlorobiphenyls

Yarek Hrywna,1,2,dagger Tamara V. Tsoi,1,3,* Olga V. Maltseva,1 John F. Quensen III,1,3 and James M. Tiedje1,2,3,*

Center for Microbial Ecology,1 Department of Microbiology,2 and Department of Crop and Soil Sciences,3 Michigan State University, East Lansing, Michigan 48824-1325

Received 25 September 1998/Accepted 11 March 1999

Cloning and expression of the aromatic ring dehalogenation genes in biphenyl-growing, polychlorinated biphenyl (PCB)-cometabolizing Comamonas testosteroni VP44 resulted in recombinant pathways allowing growth on ortho- and para-chlorobiphenyls (CBs) as a sole carbon source. The recombinant variants were constructed by transformation of strain VP44 with plasmids carrying specific genes for dehalogenation of chlorobenzoates (CBAs). Plasmid pE43 carries the Pseudomonas aeruginosa 142 ohb genes coding for the terminal oxygenase (ISPOHB) of the ortho-halobenzoate 1,2-dioxygenase, whereas plasmid pPC3 contains the Arthrobacter globiformis KZT1 fcb genes, which catalyze the hydrolytic para-dechlorination of 4-CBA. The parental strain, VP44, grew only on low concentrations of 2- and 4-CB by using the products from the fission of the nonchlorinated ring of the CBs (pentadiene) and accumulated stoichiometric amounts of the corresponding CBAs. The recombinant strains VP44(pPC3) and VP44(pE43) grew on, and completely dechlorinated high concentrations (up to 10 mM), of 4-CBA and 4-CB and 2-CBA and 2-CB, respectively. Cell protein yield corresponded to complete oxidation of both biphenyl rings, thus confirming mineralization of the CBs. Hence, the use of CBA dehalogenase genes appears to be an effective strategy for construction of organisms that will grow on at least some congeners important for remediation of PCBs.

* Corresponding author. Mailing address: A540 Plant and Soils Sciences Building, Center for Microbial Ecology, Michigan State University, East Lansing, MI 48824-1325. Phone: (517) 353-7858; (517) 432-1536. Fax: (517) 353-2917. E-mail: tiedjej{at}pilot.msu.edu; tsoi{at}pilot.msu.edu.

dagger Present address: The Ecosystems Center, Marine Biological Labs, Woods Hole, MA 02543-1015.

Applied and Environmental Microbiology, May 1999, p. 2163-2169, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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