Suicide Inactivation of Catechol 2,3-Dioxygenase from Pseudomonas putida mt-2 by 3-Halocatechols.

Abstract:

:The inactivation of catechol 2,3-dioxygenase from Pseudomonas putida mt-2 by 3-chloro- and 3-fluorocatechol and the iron-chelating agent Tiron (catechol-3,5-disulfonate) was studied. Whereas inactivation by Tiron is an oxygen-independent and mostly reversible process, inactivation by the 3-halocatechols was only observed in the presence of oxygen and was largely irreversible. The rate constants for inactivation (K(2)) were 1.62 x 10 sec for 3-chlorocatechol and 2.38 x 10 sec for 3-fluorocatechol. The inhibitor constants (K(i)) were 23 muM for 3-chlorocatechol and 17 muM for 3-fluorocatechol. The kinetic data for 3-fluorocatechol could only be obtained in the presence of 2-mercaptoethanol. Besides inactivated enzyme, some 2-hydroxyhexa-2,4-diendioic acid was formed from 3-chlorocatechol, suggesting 5-chloroformyl-2-hydroxypenta-2,4-dienoic acid as the actual suicide product of meta-cleavage. A side product of 3-fluorocatechol cleavage is a yellow compound with the spectral characteristics of a 2-hydroxy-6-oxohexa-2,4-dienoic acid indicating 1,6-cleavage. Rates of inactivation by 3-fluorocatechol were reduced in the presence of superoxide dismutase, catalase, formate, and mannitol, which implies that superoxide anion, hydrogen peroxide, and hydroxyl radical exhibit additional inactivation.

journal_name

Appl Environ Microbiol

authors

Bartels I,Knackmuss HJ,Reineke W

doi

10.1128/AEM.47.3.500-505.1984

keywords:

subject

Has Abstract

pub_date

1984-03-01 00:00:00

pages

500-5

issue

3

eissn

0099-2240

issn

1098-5336

journal_volume

47

pub_type

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