Cloning and expression in Escherichia coli of Pseudomonas strain LB400 genes encoding polychlorinated biphenyl degradation

Journal Article · Wed Mar 01 04:00:00 EST 1989 · J. Bacteriol.; (United States)

OSTI ID:6071918

Mondello, F J

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. This 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.

• Research Organization: Research and Development Center, GE Co., Schenectady, NY (USA)
• OSTI ID: 6071918
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 171:3; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AROMATICS
BACTERIA
BIODEGRADATION
BIOLOGICAL PATHWAYS
CELL CONSTITUENTS
CELL CULTURES
CHEMICAL REACTIONS
CHLORINATED AROMATIC HYDROCARBONS
CLONE CELLS
COSMIDS
DECOMPOSITION
ENZYMES
ESCHERICHIA COLI
GENE REGULATION
GENES
GROWTH
HALOGENATED AROMATIC HYDROCARBONS
MICROORGANISMS
ORGANIC CHLORINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
OXIDOREDUCTASES
OXYGENASES
PLASMIDS
PSEUDOMONAS