WAL lysin purfication - streamlined summary

WAL lysin purfication

PBCV1 WAL lysin was prepared from a 20x conc., 1500ml cell culture with MES buffer, w/o mercaptoethanol.

Column is Pall Acrosep Q HyperD (quaternary ammonium). Five ml of sample of PBCV WAL lysin was applied. The column was eluted by step gradients of ammonium acetate (2ml each) of 0.05, 0.1, 0.2M. Fractions 6-10 are 0.1M ammonium acetate.

10 microliter aliquots of column fractions were added to 100 microliters of cells suspended in MES plus 2mM CaCl2 and 0.5% Triton X-100. Suspensions were centrifuged 30" at 4500 RPM after incubating 30' at 37C.

Lysin activity of WAL lysin column fractions

SDS PAGE of column fractions