Activation of streptolysin S-related cellular potential hemolysin by grinding with glass beads

Abstract

Cellular potential hemolysin, the precursor to extracellular streptolysin S, was activated by grinding with glass beads on a Vortex mixer. Hemolysin activity was markedly enhanced by the addition of ribonucleic acid-core during the grinding. Determinants for optimum results with the beads included small bead size, prewashing of beads with acetic acid, and appropriate bead volume during grinding. The increased sensitivity of this technique over those previously used revealed cellular potential hemolysin in cells grown in the presence of glucose, which inhibits streptolysin S production. The basis for the increased sensitivity of this procedure may be due to the binding of cellular potential hemolysin or ribonucleic acid to the beads.

Membrane and cytoplasmic location of streptolysin S precursor

“…However, little CPH was recoverable from the supernatant after sedimentation of the residual membrane by centrifugation at 100,000 x g for 90 min. Thus far, the only way to free active hemolysin from its CPH form is by sonication (5) or blending in a Vortex mixer with glass beads (7).…”

Vortex grinding for mechanochemistry: application for automated supramolecular catalysis and preparation of a metal–organic framework

“…leaf tissue) to afford fine powders within periods of minutes. 3 The vortex mixer has also been used for the solvent-free preparation of insoluble polymers for MALDI mass spectrometry. The method involved adding dry components of a sample to a small glass vial with two metal balls (or BBs) made of steel and Zn plated.…”