Protoplast membrane of Streptococcus faecalis

Shockman, Gerald D. (Temple University School of Medicine, Philadelphia, Pa.), Joseph J. Kolb, Bohdan Bakay, Margaret J. Conover, and Gerrit Toennies. Protoplast membrane of Streptococcus faecalis. J. Bacteriol. 85:168-176. 1963.-The membrane fraction of Streptococcus faecalis (ATCC 9790) was isolated and purified, by a variety of procedures, from cultures that were grown under closely controlled conditions of physiological age and nutrition. The most satisfactory method required the use of lysozyme-to-cell ratios below 0.01 and the intermediate formation of protoplasts in osmotically protective media. Amino acid analyses of three of the membrane preparations indicated a characteristic and constant, but not unusual, pattern; 42% of the membranes from threonine-depleted and 49 to 55% of the membranes from log-phase cultures were accounted for as protein. Significant quantities of d-alanine or d-aspartic acid were not detected, indicating the absence of contaminating cell-wall substance. Essentially, all of the nitrogen was accounted for as amino acids. The lipid content of membranes from stationary-phase threonine-depleted (36%) and valine-depleted (40%) cultures was significantly higher than the corresponding fraction of exponential-phase cultures (28%). The phosphorus content of the membrane lipid was relatively constant (2.8 to 3.0%), and the nitrogen content was extremely low (0.12 to 0.26%). Thus, changes in the composition of the membrane fraction occurred during the transition of log-phase cells into threonine- or valine-depleted cells.