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Chemical composition and serology of Mycoplasma pneumoniae lipids.

M. pneumoniae strain 65-2053 was grown on glass in a buffered medium containing PPLO broth (Difco), 3% PPLO Serum Fraction (Difco), and 5 tg of oleic acid per ml, as we have previously described [8]. Cells were harvested before the pH of the overlaying culture fluids reached 6.5. Membranes were obtained by osmotic lysis [9]. To obtain total cellular lipid, washed cells were lypholized and extracted four times with chloroform-methanol (2:1). The pooled organic residues were freed of nonlipids by passage through Sephadex G-25 following the procedure of Siakotos and Rouser [10]. Immunologic, serologic, and other procedures and reagents have been previously described [9]. Table 1 presents a comparative study of the amount of total lipid recovered as the percentage of dry weight of M. pneumoniae and Acholeplasma laidlawii B. A. laidlawii B was grown in deep culture, but in the same medium. We also show, for comparison, values found in the available literature. We found about 26% of the dry weight of M. pneumoniae to be lipid. Others using a different strain and medium found considerably less lipid, about 12%. Comparison of our data for M. pneumoniae with that reported for other