Evaluation of the potential of a bead mill for the release of intracellular bacterial enzymes

The suitability of bead mills for the release of intracellular bacterial enzymes has been studied using the Dyno-Mill Model KDL. The effect of cell concentration, bead size and agitation speed on the release of beta-lactamase from Enterobacter cloacae P99 was examined. Scale-up considerations included, the best operational values for these parameters were 1 g cell paste suspended in 2.5 ml buffer, 0.25 mm diameter glass beads and 15 ms ⁻¹ agitation speed. These conditions proved suitable for the release of enzymes from other Gram-negative bacteria in both batch and continuous processes.