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Biophys J, April 2000, p. 1748-1764, Vol. 78, No. 4
Departments of *Chemistry and #Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA
To obtain turgor pressure, intracellular osmolalities,
and cytoplasmic water activity of Escherichia coli as a
function of osmolality of growth, we have quantified and analyzed
amounts of cell, cytoplasmic, and periplasmic water as functions of
osmolality of growth and osmolality of plasmolysis of nongrowing cells
with NaCl. The effects are large; NaCl (plasmolysis) titrations of cells grown in minimal medium at 0.03 Osm reduce cytoplasmic and cell
water to ~20% and ~50% of their original values, and increase periplasmic water by ~300%. Independent analysis of amounts of cytoplasmic and cell water demonstrate that turgor pressure decreases with increasing osmolality of growth, from ~3.1 atm at 0.03 Osm to
~1.5 at 0.1 Osm and to less than 0.5 atm above 0.5 Osm. Analysis of
periplasmic membrane-derived oligosaccharide (MDO) concentrations as a
function of osmolality, calculated from literature analytical data and
measured periplasmic volumes, provides independent evidence that turgor
pressure decreases with increasing osmolality, and verifies that
cytoplasmic and periplasmic osmolalities are equal. We propose that MDO
play a key role in periplasmic volume regulation at low-to-moderate
osmolality. At high growth osmolalities, where only a small amount of
cytoplasmic water is observed, the small turgor pressure of E. coli demonstrates that cytoplasmic water activity is only slightly
less than extracellular water activity. From these findings, we deduce
that the activity of cytoplasmic water exceeds its mole fraction at
high osmolality, and, therefore, conclude that the activity coefficient
of cytoplasmic water increases with increasing growth osmolality and
exceeds unity at high osmolality, presumably as a consequence of
macromolecular crowding. These novel findings are significant for
thermodynamic analyses of effects of changes in growth osmolality on
biopolymer processes in general and osmoregulatory processes in
particular in the E. coli cytoplasm.
Biophys J, April 2000, p. 1748-1764, Vol. 78, No. 4
© 2000 by the Biophysical Society 0006-3495/00/04/1748/17 $2.00
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