Evidence that dipicolinic acid is covalently bound to specific macromolecules in spores of Bacillus subtilis

Two dipicolinic acid (DPA)-binding macromolecules with molecular masses of about 440 kDa and 230 kDa were detected in the soluble fractions of dormant and germinated spores of Bacillus subtilis using native PAGE and an immunological technique. In SDS-PAGE, only one band with the molecular mass of about 50 kDa was found. Proteinase K partially digested the 440-kDa macromolecule of dormant spores to convert it into a 230-kDa one, and completely digested both the 440-kDa and 230-kDa bands of germinated spores. DNase I did not affect either DPA-binding macromolecules. This suggests that the two DPA-binding macromolecules are of similar origin, their main component is protein and a conformational change may occur during germination. DPA was not dissociated from the DPA-binding macromolecules by extensive dialysis and SDS treatment, suggesting the presence of a covalent bonding.