Sauer:Lysing E. coli with Lysozymes: Difference between revisions

Native lysis is a staple protocol in practically every biochemistry lab, yet there is significant variability in the procols I have read.  The intention is to liberate the guts of E. coli without disturbing the native conformations of the biomolecules.  Using a French pressure cell you can sheer the cells open, this is a great way to lyse cells.  Unfortunately, it is impractical for multiple samples or small-volume samples that don't fit in the pressure cell.  Besides, the machine is scary, lysis can be variable, and cleanup is a hassle.  Chickens and bacteriophage have evolved great a way of opening E. coli using enzymes.  Most commercial lysozymes are free from proteolytic activity and can be added in large amounts.  Be careful, if your protein is ~14 kDa you may inadvertantly purify the added lysozyme instead of your target protein.  Don't laugh, it's happened more than once and I have met people who solved the crystal stucture of lysozyme by mistake.