No abstract

Spheroplasts of Mycobacterium tuberculosis were formed by culture of the organism in a nutritionally complete medium to which lysozyme and an extract obtained from rabbit peritoneal monocytes were added. The potentiating effect of the factor, or factors, present in the monocyte extract was lost upon prolonged storage at −20 °C or heat at 90 °C for 5 minutes, but was unaffected by dialysis. It was inactive in the presence of fluoride or oxalate. Enzymatic analysis of the monocyte extract showed the presence of lysozyme, acid phosphatase, phospholipase, and trypsin. When acid phosphatase or phospholipase were substituted for the monocyte extract in the lysozyme test system, spheroplast formation was obtained.

Lysozyme in combination with ethylenediamine tetraacetic acid (EDTA) gives rise to spheroplasts when Mycobacterium tuberculosis is cultured in a nutritionally complete Tween 80 – albumin medium. These spheroplasts do not multiply in this medium, but when such forms, stabilized by the presence of sucrose and Mg++, are inoculated into PPLO agar or broth an L-form of growth is initiated. A CO2requirement was demonstrated by L-forms grown on solid media.

No abstract