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J. Bacteriol., 11 1995, 6176-6183, Vol 177, No. 21
R Merchante, HM Pooley and D Karamata
The possibility of there being a periplasm in Bacillus subtilis, in the
distinct cell compartment bounded by the cytoplasmic membrane and the thick
cell wall, has been investigated quantitatively and qualitatively.
Cytoplasmic, membrane, and protoplast supernatant fractions were obtained
from protoplasts which were prepared isotonically from cells grown under
phosphate limitation. The contents of the protoplast supernatant fraction
represent an operational definition of the periplasm. In addition, this
cell fraction includes cell wall-bound proteins, exoproteins in transit,
and contaminating cytoplasmic proteins arising through leakage from, or
lysis of a fraction of, protoplasts. The latter, measured by assay of
enzyme markers and by radiolabeled RNA and protein, was found to represent
7.6% of total cell protein, yielding a mean of 9.8% +/- 4.8% for B.
subtilis 168 protein considered periplasmic. Qualitatively, after
subjection of all cell fractions to polyacrylamide gel electrophoresis,
RNase and DNase, zymographs revealed that (i) each cell fraction had a
unique profile of nucleases and (ii) multiple species and a major fraction
of both nucleases were concentrated in the periplasm. We conclude that the
operationally defined periplasmic fraction corresponds closely, both
quantitatively and qualitatively, to the contents of the periplasm of
Escherichia coli. We discuss evidence that the maintenance of the
components of this surface compartment in B. subtilis is compatible with
the thick negatively charged cell wall acting as an external permeability
barrier.
Copyright © 1995, American Society for Microbiology
A periplasm in Bacillus subtilis
Institut de Genetique et de Biologie Microbiennes, Lausanne, Switzerland.
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