Question

Why lysozyme is added to cell pellet before sonication?

Answer 1

Hi, 

I am trying to lyse gram positive E. faecalisusing lysozyme (final concentration: 100ug/ml). I suspend the bacterial pellet with lysozyme in buffer for 30min at 37C. Then run the supernatant on SDS-PAGE. I do see many protein bands. I am not sure if these are the proteins leaked from the cytoplasm or from the peptidoglycan layer. So my question is to know if lysozyme can be used to break open the gram positive bacteria to release cytoplasmic content?

Buffer composition: 0.2mM EDTA, 50mM Tris pH 8, 10% glycerol, 1mM PMSF, 0.1% Triton X100, 150mM NaCl