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A periplasm in Bacillus subtilis

R Merchante1, H M Pooley, D Karamata

  • 1Institut de Génétique et de Biologie Microbiennes, Lausanne, Switzerland.

Journal of Bacteriology
|November 1, 1995
PubMed
Summary
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Bacillus subtilis possesses a periplasm, a cell compartment containing 9.8% of its protein, including essential nucleases. This finding supports its structural similarity to Escherichia coli.

Area of Science:

  • Microbiology
  • Cell Biology
  • Biochemistry

Background:

  • The existence and composition of the periplasm in Bacillus subtilis, a Gram-positive bacterium, remain areas of investigation.
  • Understanding the periplasm is crucial for comprehending bacterial cell structure and function, particularly in relation to the cell wall.

Purpose of the Study:

  • To quantitatively and qualitatively investigate the presence and contents of the periplasm in Bacillus subtilis.
  • To compare the periplasmic fraction of Bacillus subtilis with that of Escherichia coli.

Main Methods:

  • Protoplasts were isolated from Bacillus subtilis cells grown under phosphate limitation.
  • Cellular fractions (cytoplasmic, membrane, protoplast supernatant) were analyzed.
  • Enzyme markers, radiolabeled RNA and protein, and polyacrylamide gel electrophoresis with zymography were employed.

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Main Results:

  • The protoplast supernatant fraction, operationally defined as the periplasm, constituted approximately 9.8% of total cell protein.
  • This fraction contained cell wall-bound proteins, exoproteins, and some contaminating cytoplasmic proteins (7.6%).
  • Nucleases (RNase and DNase) showed unique profiles in each fraction, with significant concentration in the periplasm.

Conclusions:

  • The operationally defined periplasmic fraction of Bacillus subtilis closely resembles that of Escherichia coli in both quantity and quality.
  • The thick, negatively charged cell wall of Bacillus subtilis likely acts as a permeability barrier, facilitating the maintenance of periplasmic components.