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Journal of Bacteriology, December 2001, p. 7182-7189, Vol. 183, No. 24
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.24.7182-7189.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Hydrogen Peroxide Fluxes and Compartmentalization
inside Growing Escherichia coli
Lauren Costa
Seaver and
James A.
Imlay*
Department of Microbiology, University of
Illinois, Urbana, Illinois 61801
Received 9 July 2001/Accepted 20 September 2001
Escherichia coli generates about 14 µM hydrogen
peroxide (H2O2) per s when it grows
exponentially in glucose medium. The steady-state intracellular
concentration of H2O2 depends on the rates at
which this H2O2 is dissipated by scavenging
enzymes and by efflux from the cell. The rates of
H2O2 degradation by the two major scavenging enzymes, alkyl hydroperoxide reductase and catalase, were quantified. In order to estimate the rate of efflux, the permeability coefficient of membranes for H2O2 was determined. The
coefficient is 1.6 × 10
3 cm/s, indicating that
permeability is substantial but not unlimited. These data allowed
internal H2O2 fluxes and concentrations to be
calculated. Under these growth conditions, Ahp scavenges the majority
of the endogenous H2O2, with a small fraction
degraded by catalase and virtually none persisting long enough to
penetrate the membrane and exit the cell. The robust scavenging
activity maintains the H2O2 concentration
inside glucose-grown cells at <10
7 M, substantially
below the level (10
6 M) at which toxicity is evident.
When extracellular H2O2 is present, its flux
into the cell can be rapid, but the internal concentration may still be
an order of magnitude lower than that outside. The presence of such
gradients was confirmed in experiments that revealed different degrees
of oxidative stress in cocultured scavenger-deficient mutants. The
limited permeability of membranes to H2O2
rationalizes the compartmentalization of scavenging systems and
predicts that bacteria that excrete redox-cycling drugs do not
experience the same H2O2 dose that they impose
on their competitors.
*
Corresponding author. Mailing address: Department of
Microbiology, University of Illinois, Urbana, IL 61801. Phone: (217) 333-5812. Fax: (217) 244-6697. E-mail: jimlay{at}uiuc.edu.
Journal of Bacteriology, December 2001, p. 7182-7189, Vol. 183, No. 24
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.24.7182-7189.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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