Ultrastructural and Chemical Alteration of the Cell Envelope of Pseudomonas aeruginosa , Associated with Resistance to Ethylenediaminetetraacetate Resulting from Growth in a Mg ²⁺ -Deficient Medium

Abstract

Cells of Pseudomonas aeruginosa became resistant to the lytic effect of ethylenediametetraacetate (EDTA) when grown in a Mg2+-deficient medium. To correlate ultrastructural changes in the cell wall associated with the shift to EDTA-resistance, a freeze-etch study was performed. Upon fracturing, the outer cell wall membrane split down the hydrophobic center to reveal the outer (concave) and inner (convex) layers. The concave cell wall layer of EDTA-sensitive cells grown in Mg2+-sufficient medium contained spherical units resting on an underlying smooth support layer. Upon EDTA treatment, approximately one-half of these spherical units were extracted. Cells grown in Mg2+-deficient medium were resistant to EDTA. The concave cell wall layer of EDTA-resistant cells had increased numbers of highly compacted spherical units, giving this layer a disorganized appearance. The highly compacted appearance of this layer was unaltered by EDTA treatment. Thus, growth in Mg2+-deficient medium resulted in cells which were resistant to EDTA and which possessed an ultrastructurally altered outer layer of the outer cell wall membrane. Cell envelopes from EDTA-resistant cells were found to possess 18% less phosphorus, 16.4% more total carbohydrate, and 13.3% more 2-keto-3-deoxyoctonate than cell envelopes from EDTA-sensitive cells. There were also qualitative, but not quantitative, differences in the protein content of cell envelopes from EDTAresistant and EDTA-sensitive cells. MATERIALS AND METHODSPreparation of chemically defined Mg2+-sufficient and Mg2+-deficient media. A basal medium (BM 1) was used as described previously by Brown and Melling (4), with the modification that the glucose concentration was raised to 0.05 M. Other 302 on August 1, 2020 by guest

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