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Abstract
Escherichia coli is able to grow with a high rate under anaerobic conditions upon decrease in redox potential (Eh) both either in
slightly alkaline (pH 7.5) or acidic (pH 5.5) medium. Upon transition of E. coli MC4100 culture to stationary growth phase a
decrease in Eh from the positive values of +120 to +160 mV to the negative ones of 380 to 550 mV, and the H2 production
are observed at various pH. A redox reagent DL -dithiothreitol (DTT) in a concentration of 3 mM reduces Eh to the negative values,
and increases a latent (lag) growth phase duration, as well as delays a logarithmic growth phase independently of pH. At alkaline
and acidic pH the changes in membrane potential (DW) are observed in the presence of 3 mM DTT. K+ uptake is recovered. At pH
5.5 the H2 production is suppressed by DTT only in a higher concentration of 10 mM. The results suggest DTT eects that are in
addition to the eects of Eh. The mechanism of DTT action on bacterial growth might be intermediated through thiol group modulation of the membrane proteins, which is reected as the generation of DW as well as K+ accumulation and the activity of the
membrane-associated enzymes.
2004 Elsevier Inc. All rights reserved.
Keywords: Redox potential;
DL -Dithiothreitol;
Bagramyan),
0006-291X/$ - see front matter 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.bbrc.2004.10.119
positive values, created by the presence of other chemicals, are not able to aect the growth [2,7]. However, E.
coli is likely to sense Eh independently of oxygen concentration and of the oxidizers [7]; dierent mechanisms and
models are proposed to explain such a redox taxis [8].
Eh might determine an electron transfer within bacterial membrane [7] and proton-motive force [9]. It is suggested that the eect of Eh on proton-motive force under
anaerobic conditions is induced by the change in pH
gradient across the membrane. The latter resulted from
an alteration in the cytoplasmic pH by fermentation
acids [10,11]; the membrane potential (DW) is changed
slightly [9]. Such a dependence is due to change in the
membrane proton permeability without modication
of the proton-translocating F0F1-ATPase activity
[9,12]. The change in the membrane proton permeability
might depend on thiol groups state and distribution
[11,13,14]. At the same time, a correlation between Eh
and the state and distribution of thiol groups on the bac-
804
G. Kirakosyan et al. / Biochemical and Biophysical Research Communications 325 (2004) 803806
G. Kirakosyan et al. / Biochemical and Biophysical Research Communications 325 (2004) 803806
805
H2 production rate
(mV Eh/min.mg d.wt)
5
-DTT
+DTT
4
3
2
1
0
pH 7.5
pH 6.5
160
pH 5.5
-DTT
+DTT
150
140
130
120
110
100
pH 7.5
pH 6.5
pH 5.5
C
Fig. 1. Eects of DTT on the growth characteristics of E. coli MC
4100 under anaerobic conditions on glucose at various pH. (A) Lag
growth phase duration. (B) Specic growth rate. Bacteria were growing
in salt medium; glucose was in concentration of 0.2%, DTT (3 mM)
was introduced into the growth medium immediately before inoculation of bacteria.
0.2
0.15
-DTT
+DTT
0.1
0.05
0
pH 7.5
pH 6.5
pH 5.5
806
G. Kirakosyan et al. / Biochemical and Biophysical Research Communications 325 (2004) 803806
Concluding remarks
It is assumed that DTT as a reducer itself aects the
growth of E. coli, as well its eects can be intermediated
through Eh and depend on external pH. It is clear that
the oxidationreduction processes play an exclusive role
in the habitability of bacteria. The majority of these processes implemented on bacterial membrane depend on
Eh and play an essential role in metabolism
[12,14,16,17]. For instance, the reduced environment
changes metabolic uxes resulting in decreased or enhanced fermentation products [2123]. But not always,
as it follows from our results (see Figs. 13) this link is
unambiguous and determines the growth of bacteria.
So, the mechanisms of Eh aecting metabolism remain
little known. In this respect it is required to further study
the eects of dierent oxidizers and reducers, which will
allow understanding regulatory paths for the membrane
functions and the mechanisms of bacterial redox taxis.
Acknowledgments
This work is fullled by the support of the Ministry
of Education and Science of the Republic of Armenia
and partially by the US Civilian Research and Development Foundation Grant No. AB1-2307-YE02.
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