Simultaneous cultivation and disruption of Escherichia coli using glass beads to release recombinant alpha-amylase and other enzymes

An efficient and reproducible protein recovery procedure from Escherichia c oli is presented. Recombinant strains overexpressing recombinant alpha-amyl ase were cultured in shaken flasks containing small glass beads. Most of th e cells underwent mechanical lysis and more than 90% of the alpha-amylase c ontent and also of other non-recombinant enzymes were released into the cul ture fluid after 24 h. The degree to which the enzymes were released depend ed on the enzymes and strains involved.