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Main Title Cloning and Expression in 'Escherichia coli' of Pseudomonas Strain LB400 Genes Encoding Polychlorinated Biphenyl Degradation.
Author Mondello, F. J. ;
CORP Author General Electric Co., Schenectady, NY. Biological Sciences Lab.;Environmental Protection Agency, Cincinnati, OH. Risk Reduction Engineering Lab.
Publisher c1989
Year Published 1989
Report Number EPA-R-812727; EPA/600/J-93/300;
Stock Number PB93-229227
Additional Subjects Biodegradation ; Polychlorinated biphenyls ; Bacteria ; Cloning ; Genetics ; Recombinant DNA ; Genetic recombination ; Molecular biology ; Biochemistry ; Microbiology ; Chemical reactions ; Bioassay ; Biological pathways ; Reprints ; Escherichia coli ; Strains(Biology) ; Pseudomonas
Library Call Number Additional Info Location Last
NTIS  PB93-229227 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 10p
Pseudomonas strain LB400 is able to degrade an unusually wide variety of polychlorinated biphenyls (PCBs). A genomic library of LB400 was constructed by using the broad-host-range cosmid pMMB34 and introduced into Escherichia coli. Approximately 1,600 recombinant clones were tested, and 5 that expressed 2,3-dihydroxybiphenyl dioxygenase activity were found. The enzyme is encoded by the bphC gene of the 2,3-dioxygenase pathway for PCB-biphenyl metabolism. Two recombinant plasmids encoding the ability to transform PCBs to chlorobenzoic acids were identified, and one of these, pGEM410, was chosen for further study. The PCB-degrading genes (bphA, -B, -C, and -D) were localized by subcloning experiments to a 12.4-kilobase region of pGEM410. The ability of recombinant strains to degrade PCBs was compared with that of the wild type. In resting-cell assays, PCB degradation by E. coli strain FM4560 (containing a pGEM410 derivative) approached that of LB400 and was significantly greater than degradation by the original recombinant strain. High levels of PCB metabolism by FM4560 did not depend on the growth of the organism on biphenyl, as it did for PCB metabolism by LB400. When cells were grown with succinate as the carbon source, PCB degradation by FM4560 was markedly superior to that by LB400. (Copyright (c) 1989, American Society for Microbiology.)