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From the
1 From the National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Maryland 20014
A number of degradative enzymes are specifically released from exponentially growing Escherichia coli by osmotic shock. They include alkaline phosphatase, cyclic phosphodiesterase, 5'-nucleotidase, acid phosphatase, and the ribonucleic acid-inhibited endonuclease that is active with deoxyribonucleic acid. Small amounts of RNase are also set free, but some 16 control enzymes are found to remain within the cell. Only about 4% of the cellular protein is released into the shock medium. The procedure for osmotic shock has been modified in order to obtain about 80% viability with exponential cells, with good release of enzymes. In the first stage, cells are exposed to 20% sucrose-1 x 10-4 M EDTA. In the second stage, the pellet of sucrose-treated cells is rapidly dispersed in cold 5 x 10-4 M MgCl2, which causes the enzymes to be liberated. At least 90% of the DNA endonuclease and 50% of the latent ribonuclease are released from exponentially growing E. coli when as much as 2 x 10-2 M MgCl2 is used in the procedure for osmotic shock.
The Release of Enzymes by Osmotic Shock from Escherichia coli in Exponential Phase
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