Cell Wall Breakage

[Jessie Micales]

>I would like to break through the cell walls of a Penicillium sp. to
>rescue a periplasmic enzyme.  I have used a number of enzymes to
>digest the cell wall.  Recently we read about a method in which the
>mycelia is lyophilized and then treated with diluted ammonium
>hydroxide.  Has anyone tried this?  Are there any better techniques?
>Thanks,
>Ted Crusberg
>Biology & Biotechnology
>Worcester Polytechnic Institute

Lyophilization can denature some enzymes.  I've never had much luck with it 
myself.  I usually use a mortar and pestle to crush hyphae in liquid 
nitrogen, perhaps with some glass beads (although they usually aren't 
necessary).  This seems to work sufficiently for what I do (enzyme 
extractions from basidiomycetes).

Jessie Micales
Forest Products Laboratory
Madison, WI