Abstract
A method for the preparation of spheroplasts from Selenomonas ruminantium is described. The method is based on the lysozyme/EDTA method developed by Kaback and collaborators. The most important modification introduced consisted in the substitution of nitrilotriacetic acid for EDTA. The final yield of spheroplasts was ≈80% of the original whole cell suspension. However, to achieve this yield, the inclusion of n-decanoic acid and higher yeast extract levels in the growth medium, as well as the substitution of mutanolysin for lysozyme and of nitrilotriacetic acid for EDTA during the incubation of cell suspensions at lower temperatures, were also all found necessary. Right-side-out vesicles were obtained from these spheroplasts according to the original method.
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