Genetic and Biochemical Characterization of the Broad Spectrum Chlorobenzene Dioxygenase from Burkholderia Sp. Strain PS12 — Dechlorination of 1,2,4,5‐Tetrachlorobenzene
Open Access
- 16 July 1997
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 247 (1) , 190-199
- https://doi.org/10.1111/j.1432-1033.1997.00190.x
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 α‐subunit and β‐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 18O2 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).Keywords
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