Effect of space factors on Escherichia coli B/r cells

Stationary phase cells of Escherichia coli B/r were inactivated when they were exposed to high vacuum (10(-6) torr). About 5% of the cells were still able to form a colony after 45 min exposure. Vacuum dried cells (0.5 torr, 120 min) show colony forming ability of 30% or more. They were inactivated to about 5% by further vacuum treatment. Vacuum treated cells showed higher permeability for various cell components. UV irradiated E. coli B/r cells in vacuum showed increased UV sensitivity. DNA-protein cross-links were preferentially formed in a vacuum. To obtain 63% of DNA cross linked with protein required 852 erg mm-2 (D37) of UV irradiation in suspension and only 72 erg mm-2 of UV irradiation in vacuum. The protein components of DNA-protein cross-links were hydrolysed with pronase E and the amino acids directly bonding to DNA were determined. The most important amino acids concerned in DNA-protein cross-links seem to be glycine and alanine, followed by aspartic acid (asparagine), glutamic acid (glutamine) and histidine. The sensitivity to X-rays of stationary phase E. coli B/r cells seems to depend on the remaining gas atmospheric in the vacuum since it varies with different pumping systems.