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PREPARATION OF SPHEROPLASTS FROM VIBRIO COMMA

Department of Microbiology, Baylor University College of Medicine, Houston, Texas

ABSTRACT

CHATTERJEE, B. R. (Baylor University College of Medicine, Houston, Texas), AND ROBERT P. WILLIAMS. Preparation of spheroplasts from Vibrio comma. J. Bacteriol. 85:838–841. 1963.—Spheroplasts were prepared from several strains of Vibrio comma by lysozyme treatment combined with freezing and thawing of the organisms. The optimal concentration of lysozyme was 50 µg/ml, although some spheroplasts formed at a concentration of 10 µg/ml. Higher concentrations (200 µg/ml) caused lysis of cells along with spheroplast formation. Treatment was carried out in broth cultures containing 15% sucrose, and if the osmotic tension was lowered the spheroplasts lysed. Some motile, spherical cells were present in every preparation. Addition of 3% glycine to broth cultures resulted in rapid transformation of the vibrios into large, spherical bodies. However, these were actively motile, and were not sensitive to a lower osmotic tension. Therefore, they could not be considered as spheroplasts.