Sensitivity to ampicillin and cephalothin of enzymes involved in wall peptide crosslinking in Escherichia coli K12, strain 44.
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Biomedical subjects
Publications and source records attributed to J Coyette.
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The autolytic N-acetylmuramidase present in Lactobacillus acidophilus strain 63 AM Gasser has an optimal pH between 5 and 6 when lysing intact cells or isolated cell walls. Cellular lysis at pH 5 is two to four times more rapid in citrate buffer of 0.01 M and 0.5 M or higher than in 0.1 M acetate buffer. It seems that sulfhydryl groups are required for both cell and wall autolysis. Heavy metal ions and p-chloro-mercuribenzoate, at low concentrations, are powerful inhibitors. Ethylenediaminetetraacetic acid stimulates cellular but not wall autolysis in acetate buffer to the level obtained in citrate buffer. The possible involvement of sulfhydryl groups in a mechanism of control of cellular autolytic activity is discussed. The autolytic enzyme, although unstable in solution at 37 C, can be extracted from walls by the use of solutions of bovine serum albumin (100 mug/ml) in 0.01 N NaOH. Soluble enzyme extracted from walls rebinds on to sodium decylsulfate-treated walls, but three times as much of the wall material is required to completely re-adsorb the activity.
Ultrastructural changes which occur during cellular autolysis of Lactobacillus acidophilus strain 63AM Gasser in 0.05 M citrate buffer, pH 5.0, were examined. Early in the process, randomly distributed electron-dense patches were seen on the wall surface, along with an accompanying eversion of mesosomes. Later, after a loss of about 20% of the initial cellular turbidity, dissolution from the outside of nascent cross walls was seen. This observation was related to the normal process of cell separation. After this stage, short lengths of the cylindrical portion of the wall appeared to be completely removed in a random manner over the entire surface. This dissolution produced gaps in the wall which allowed the extrusion of membrane and cytoplasm. Although membrane was usually extruded through one major, polar, subpolar, or septal site, other secondary points of membrane extrusion were also frequently seen in the same cell section.
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