Growth and physiological characteristics of Bacillus subtilis L-forms.

The specific growth rate of stable Bacillus subtilis L-forms was slower (mu = 0.127) than the cell-walled form (mu = 0.219) when measured by optical density (O.D.). However, the L-form growth rate increased (mu = 0.288) when determined by viable cell counts. L-forms of B. subtilis appear to enter a phase of rapid cell division, followed by a period of cell enlargement which is associated with an increase in the number of cells with vacuoles and granules. Thus, maximum viable L-form numbers and DNA content were attained at approx. 30h, before maximum protein content was achieved (46h) and before maximum O.D. was observed at 71 h. Measurements showed that L-form cell size increased even after cell division had stopped. O.D. was therefore inaccurate for assessment of L-form growth. L-forms were sensitive to osmotic shock and unlike the cell-walled organisms from which they were derived, were resistant to penicillin, indicating a loss of peptidoglycan. The L-forms were similar to cell-walled forms in that antibiotic(s) and proteases were produced.