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PMID:9249026

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Citation

Beil, S, Happe, B, Timmis, KN and Pieper, DH (1997) Genetic and biochemical characterization of the broad spectrum chlorobenzene dioxygenase from Burkholderia sp. strain PS12--dechlorination of 1,2,4,5-tetrachlorobenzene. Eur. J. Biochem. 247:190-9

Abstract

The bacterium, Burkholderia (previously Pseudomonas) sp. strain PS12, reported earlier to degrade 1,2,4-trichlorobenzene is shown here to utilize also 1,2,4,5-tetrachlorobenzene (Cl4-benzene) as a growth substrate. To investigate the possibility that this organism attacks Cl4-benzene with a chlorobenzene dioxygenase which concomitantly causes dehalogenation, and to analyze the substrate range of the initial enzyme, a 5503-bp DNA fragment from PS12, exhibiting high similarity to genes coding for class IIB dioxygenases, was cloned and expressed in Escherichia coli. The sequence includes the tec genes coding for the alpha-subunit and beta-subunit of a terminal dioxygenase, a ferredoxin and a reductase. E. coli cells producing these proteins were able to dioxygenolytically attack a range of aromatic compounds including chlorinated benzenes and toluene, and also dinuclear aromatics such as biphenyl and dibenzo-p-dioxin. The enzyme was shown by (18)O2 incorporation experiments to dioxygenolytically attack a chlorosubstituted carbon atom of Cl4-benzene, thereby forming an unstable diol intermediate which spontaneously rearomatizes with concomitant chloride elimination to the corresponding 3,4,6-trichlorocatechol (Cl3-catechol).

Links

PubMed

Keywords

Amino Acid Sequence; Base Sequence; Biodegradation, Environmental; Burkholderia/metabolism; Chlorobenzenes/metabolism; Dioxygenases; Molecular Sequence Data; Oxygenases/chemistry; Oxygenases/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

9BURK:O69264

GO:0000166: nucleotide binding

ECO:0000314:

F

fig. 1: chloride release with growth turbidity fig.4: C13-catechol production shown with degradation of CI4- benzene

complete
CACAO 4541


See also

References

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