Staphylococcus aureus MF31 can grow at 46 degrees C, 2 degrees C above its normal maximum temperature of growth if 1 M NaCl is added to the medium. In the present work we show that monosodium glutamate, proline, threonine, aspartic acid, and betaine (in order of decreasing effectiveness) also enabled cells to grow at 46 degrees C. Cells grown at 46 degrees C in he presence of salt (protected or P cells) accumulated glutamate more rapidly than cells grown at 37 degrees C without salt (normal or N cells) and contained an increased amino acid pool. The principal constituents of this pool were dicarboxylic amino acids and proline. Turbidimetric evidence suggests that NaCl caused plasmolysis in S. aureus. The P cells, although grown in 1 M NaCl, had about the same Cl- and K+ content as the N cells grown without added NaCl. P cells had increased heat resistance but high concentrations of CaCl2 in the heating menstruum reduced their D55 value from a maximum of 214 min to less than 30 s. We suggest that growth at 46 degrees C in 1 M NaCl can be explained, in part at least, by the increased amino acid pool internal to the cell and the external osmotic support given by Cl- anions excluded by the cell.

Staphylococcus aureus MF31 was grown to stationary phase in a complex medium at 30, 37, and 43 degrees C in the absence of salt and at 37 and 46 degrees C in the same medium supplemented with 1 M NaCl. The principal phospholipids were cardiolipin, phosphatidylglycerol, aminoacylphosphatidyl glycerol, mono- and di-glycosyldiglyceride, and traces of phosphoglycolipid. The proportion of cardiolipin decreased with increasing growth temperature, but only slightly in the presence of 1 M NaCl, while that of aminoacylphosphatidyl glycerol was unaffected by growth temperature in absence of salt, but was about halved in the presence of 1 M NaCl. The net negative charge per mole phospholipid was greatly increased in the presence of 1 M NaCl. In the absence of salt, temperature had no effect on the total lipid content, but cells from the 46 degrees C culture in 1 M NaCl contained 25% less total lipid. The proportion of phospholipid in the total lipids, both in the absence and presence of salt, declined with increasing growth temperature. The proportion of glycolipids, however, increased with temperature both in the absence and presence of salt. It is suggested that the increase in glycolipid content and in negative charge/mole phospholipid is a part of the adaptation of S. aureus to the combination of high temperature and 1 M NaCl giving its membrane increased stability and possibly helping to exclude Cl- anion from the cell interior.