E.coli cell breaking

[ahillar at your-servername.Lan1.UManitoba.CA]

In article <3e8mnu$cd2 at elna.ethz.ch> GRUEMSCHI at EZINFO.VMSMAIL.ETHZ.CH (EMMERLING,MARCEL) writes:
>From: GRUEMSCHI at EZINFO.VMSMAIL.ETHZ.CH (EMMERLING,MARCEL)
>Subject: E.coli cell breaking
>Date: 2 Jan 1995 11:06:38 GMT

>I want to determine intermediate products of E.coli (PEP, Pyr, G6P, etc). I
>tried to break my cells with perchloric acid and KOH treatment, but i finally
>found out, that some products are acid instable. Is there any method of
>breaking cells without acid.


>Thanx,

>        Marcel
>gru at ibtech.biol.ehtz.ch

You could try breaking your cells in a French Pressure Cell (if you can locate 
one), or perhaps try either sonication or use glass beads. These methods work 
best at relatively high cell densities, and the latter two require cooling 
between breakage cycles. Have a look at Guide to Protein Purification, a 
relatively recent Meth. Enzymol. (can't remember the volume number) edited by 
Deutscher; it gives a good rundown on cell breakage methods as well as some 
protocols.

Alex