Synthesis of cell envelope components by anucleate cells (minicells) of Bacillus subtilis

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Abstract
Minicells produced by B. subtilis CU403 (divIVB1) are capable of mucopeptide biosynthesis as shown by the incorporation of L-alanine, D-alanine and N-acetylglucosamine into trichloroacetic acid [TCA]-precipitable material, which can be degraded to TCA-soluble material by lysozyme digestion. Incorporation of the precursors is sensitive to vancomycin and D-cycloserine and insensitive to chloramphenicol. Penicillin inhibits the incorporation of D- and L-alanine N-acetylglucosamine at concentrations > 10 .mu.g of penicillin/ml; however, minicells are insensitive to penicillin-induced lysis. The material synthesized in minicells from N-acetylglucosamine is not subject to turnover during a subsequent 6-h incubation period. [2-3H]glycerol is converted to a cold TCA-precipitable form by minicells. This synthesis is not inhibited by vancomycin, penicillin, D-cycloserine or chloramphenicol. Fractionation of the material synthesized from glycerol into hot TCA soluble material and chloroform/methanol-extractable material indicates that minicells convert glycerol into teichoic acid and lipid.