Release and recovery of forespores from Bacillus cereus

A method is described which makes possible the release of immature forespores from sporulating cells at specific stages of development, from the completion of stage III through to mature spore formation. With the aid of zonal density gradient centrifugation, the method makes possible the recovery of quantities of forespores ample for biochemical and physical studies. With the capability to examine forespores and some mother cell components independently, we have established that several enzymes associated with the sporulation process are localized in the newly developed forespores. Studies showed that aspartate aminotransferase and alanine aminotransferase are associated with the forespores, whereas l-alanine dehydrogenase is found only in the mother cell cytoplasm.