why EDTA is used for lysozyme action
Sudheendra Rao N R
(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.
On 11/1/07, ChenHA <hzhen from freeuk.com> wrote:
> chiranjit chowdhury wrote:
> > Dear friends, I got protocol from pharmacia for Gram negative bacterial
> > lysis with lysozyme and EDTA. I have gone through several literature
> > 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
> > 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
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