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Journal of Bacteriology, April 1999, p. 2209-2216, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Acid- and Base-Induced Proteins during Aerobic and
Anaerobic Growth of Escherichia coli Revealed by
Two-Dimensional Gel Electrophoresis
Darcy
Blankenhorn,
Judith
Phillips, and
Joan L.
Slonczewski*
Department of Biology, Kenyon College,
Gambier, Ohio 43022
Received 8 September 1998/Accepted 25 January 1999
Proteins induced by acid or base, during long-term aerobic or
anaerobic growth in complex medium, were identified in
Escherichia coli. Two-dimensional gel electrophoresis
revealed pH-dependent induction of 18 proteins, nine of which were
identified by N-terminal sequencing. At pH 9, tryptophan deaminase
(TnaA) was induced to a high level, becoming one of the most abundant
proteins observed. TnaA may reverse alkalinization by metabolizing
amino acids to produce acidic products. Also induced at high pH, but
only in anaerobiosis, was glutamate decarboxylase (GadA). The
gad system (GadA/GadBC) neutralizes acidity and enhances
survival in extreme acid; its induction during anaerobic growth may
help protect alkaline-grown cells from the acidification resulting
from anaerobic fermentation. To investigate possible responses to
internal acidification, cultures were grown in propionate, a
membrane-permeant weak acid which acidifies the cytoplasm. YfiD, a
homologue of pyruvate formate lyase, was induced to high levels at pH
4.4 and induced twofold more by propionate at pH 6; both of these
conditions cause internal acidification. At neutral or alkaline pH,
YfiD was virtually absent. YfiD is therefore a strong candidate for
response to internal acidification. Acid or propionate also increased
the expression of alkyl hydroperoxide reductase (AhpC) but only during
aerobic growth. At neutral or high pH, AhpC showed no significant
difference between aerobic and anaerobic growth. The increase of AhpC
in acid may help protect the cell from the greater concentrations of
oxidizing intermediates at low pH. Isocitrate lyase (AceA) was induced
by oxygen across the pH range but showed substantially greater
induction in acid or in base than at pH 7. Additional responses
observed included the induction of MalE at high pH and induction of
several enzymes of sugar metabolism at low pH: the phosphotransferase
system components ManX and PtsH and the galactitol fermentation enzyme
GatY. Overall, our results indicate complex relationships between pH
and oxygen and a novel permeant acid-inducible gene, YfiD.
*
Corresponding author. Mailing address: Department of
Biology, Kenyon College, Gambier, OH 43022. Phone: (740) 427-5383. Fax: (740) 427-5741. E-mail: slonczewski{at}kenyon.edu.
Journal of Bacteriology, April 1999, p. 2209-2216, Vol. 181, No. 7
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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