Selective release of enzymes from bacteria.
A group of hydrolytic enzymes, including phosphatases and nucleases, is selectively released from E. coli and certain other Gram-negative bacteria by a process designated as osmotic shock. This procedure involves exposure of the cells to ethylenediaminetetraacetate (EDTA) in 0.5 molar sucrose followed by a sudden osmotic transition to cold, dilute MgCl(2). Osmotic shock also results in an alteration of the permeability barrier of the bacterial cell and a depletion of the pool of acid-soluble nucleotides, but there is no loss of viability. On being restored to growth medium, the shocked cells recover after a lag period. Formation of spheroplasts by treatment with EDTA and lysozyme leads to selective release of the same group of enzymes. We believe that the selectively released enzymes are confined in a region between the bacterial cell wall and the cytoplasmic membrane. Histochemical studies indicate such a localization. Further, the enzyme activities are measurable with intact cells, even when the substrate is a nucleotide, to which whole cells are impermeable. Another piece of evidence concerns a mutant E. coli with a defective cell wall. In contrast to normal bacteria, this organism loses one of these enzymes into the medium in the course of growth. After osmotic shock, the bacteria show reduced uptake of sulfate,betagalactosides, galactose, and certain amino acids. Furthermore, the shock treatment causes the release of nondialyzable factors able to bind sulfate, galactose, and the same amino acids. A possible interpretation of these observations is the following: the binding proteins occupy sites near the bacterial surface, and they may be components of active transport systems responsible for the concentrative uptake of these nutrients.
D000135:Acid Phosphatase / Q000302:isolation & purification*; D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000596:Amino Acids / Q000378:metabolism; D001693:Biological Transport, Active; D002473:Cell Wall / Q000187:drug effects; D003851:Deoxyribonucleases / Q000302:isolation & purification*; D003956:Dialysis; D004492:Edetic Acid / Q000494:pharmacology*; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D009113:Muramidase / Q000494:pharmacology*; D009994:Osmolar Concentration; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification*; D011755:Pyrophosphatases / Q000302:isolation & purification*; D012260:Ribonucleases / Q000302:isolation & purification*
D000596:Amino Acids; D004492:Edetic Acid; D003851:Deoxyribonucleases; D012260:Ribonucleases; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D010744:Phosphoric Monoester Hydrolases; D009113:Muramidase; D011755:Pyrophosphatases