Cellular Destruction - (Mar/10/2009 )

Does anyone know a general solvent that would reduce a yeast cell down to its molecular components without homogenising the cell first?

i.e. completely dissolve the cell wall, membrane and internal structures with no agitation

this is probably a basic problem but i'm not a biologist so am struggling with ideas beyond strong base/acid or H2O2, any help or suggestions would be greatly appreciated

cheers

As far as I know, there is no one reagent that will completely dissolve all the cellular components of a cell. Usually the most difficult is the cell wall and/or "insoluble" proteins. Your best bet is to use a multi-reagent system such as protease, detergent and some solvent.

use zymolyase to digest the cell walls. This leaves viable cells without their cell wall.

Once the cells have spheroplased, you can then break the cells up using detergent (SDS) or other cell lysis formulations.

What exactly do you want to do? It would be helpful for us to suggest the appropriate protocol.

perneseblue on Mar 11 2009, 02:43 AM said:

i'm coating the cells with inorganic material to form a capsule, i then need to remove the cellular material and leave the capsules behind.

This is why i need to break down the cellular material to solvated molecules so they can diffuse out of the capule.

Oh and what does spheroplased mean? im a chemist with some biological knowlege but im unfamiliar with this term

And could you suggest some formulations that dont include enzymes? would repeated washings with appropriate solutions work? peroxide followed by a strong acid have been suggested to me already but im guessing this wouldn't be sufficient.

Cheers for the help so far

Try alkaline hydrolysis of the proteins, then washes with detergents and water to remove the fatty-acids and soluble components.