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J. Bacteriol., 05 1996, 2656-2661, Vol 178, No. 9
V Romanov and RP Hausinger
Corynebacterium sepedonicum KZ-4, described earlier as a strain capable of
growth on 2,4-dichlorobenzoate (G.M. Zaitsev and Y.N. Karasevich,
Mikrobiologiya 54:356-369, 1985), is known to metabolize this substrate via
4-hydroxybenzoate and protocatechuate, and evidence consistent with an
initial reductive dechlorination step to form 4-chlorobenzoate was found in
another coryneform bacterium, strain NTB-1 (W.J.J. van den Tweel, J.B. Kok,
and J.A.M. de Bont, Appl. Environ. Microbiol. 53:810- 815, 1987).
2-Chloro-4-fluorobenzoate was found to be converted stoichiometrically to
4-fluorobenzoate by resting cells of strain KZ-4, compatible with a
reductive process. Experiments with cell extracts demonstrated that Mg -
ATP and coenzyme A (CoA) were required to stimulate reductive
dehalogenation, consistent with the intermediacy of
2-chloro-4-fluoro-benzoyl-CoA and 2,4-dichlorobenzoyl-CoA thioesters.
2,4-Dichlorobenzoyl-CoA was shown to be converted to 4-chlorobenzoyl- CoA
in a novel NADPH-dependent reaction in extracts of both KZ-4 and NTB-1. In
addition to the ligase and reductive dehalogenase activities, hydrolytic
4-chlorobenzoyl-CoA dehalogenase and thioesterase activities,
4-hydroxybenzoate 3-monooxygenase, and protocatechuate 3,4- dioxygenase
activities were demonstrated to be present in the soluble fraction of KZ-4
extracts following ultracentrifugation. We propose that the pathway for
2,4-dichlorobenzoate catabolism in strains KZ-4 and NTB-1 involves
formation of 2,4-dichlorobenzoyl-CoA, NADPH- dependent ortho dehalogenation
yielding 4-chlorobenzoyl-CoA, hydrolytic removal of chlorine from the para
position to generate 4-hydroxybenzoyl- CoA, hydrolysis to form
4-hydroxybenzoate, oxidation to yield protocatechuate, and oxidative ring
cleavage.
Copyright © 1996, American Society for Microbiology
NADPH-dependent reductive ortho dehalogenation of 2,4-dichlorobenzoic acid in Corynebacterium sepedonicum KZ-4 and Coryneform bacterium strainNTB-1 via 2,4-dichlorobenzoyl coenzyme A
Center for Microbial Ecology, Michigan State University, East Lansing 48824, USA.
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