why EDTA is used for lysozyme action

Sudheendra Rao N R via methods%40net.bio.net (by sudhee26 from gmail.com)
Thu Nov 1 08:42:17 EST 2007


I doubt whether it has to do with the cell wall..probably
lipoppolysaccharide and protein outermembrane is succeptible to EDTA
disruption, than the peptidoglycan layer of gram positive bacteria.

Correct me.

Sudheendra
NBRC


On 11/1/07, ChenHA <hzhen from freeuk.com> wrote:
>
> chiranjit chowdhury wrote:
> > Dear friends, I got protocol from pharmacia for Gram negative bacterial
> cell
> > lysis with lysozyme and EDTA. I have gone through several literature
> where
> > they have used EDTA with lysozyme mediated cell lysis of Gram negative
> > bacteria but not in case of Gram positive bacteria.
> > Can anybody tell me why EDTA is used specially for Gram negative
> bacteria,
> > the exact mechanism behind it
> > Regards.
> >
> >
>
> EDTA chelates divalent cations like calcium and magnesium.  IIRC, these
> divalent cations are important for maintaining the structures on the
> cell surface.  Removing them destablises these cell surface structures
> and makes it easier to lyse the cells.
>
> As far as I can remember, having too much as well as too little calcium
> and magnesium can change the cell surface structures which, if I am
> remember correctly, is I think one reason why you use calcium when you
> prepare competent cells to make the cells more permeable to DNA (calcium
> also help screen out the charges allowing the DNA to stick to the cell
> surface).
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