I'm trying to purify my proteins which are cell wall binding domain and I found that it binds to E. coli cell wall. So when I break the cell, my proteins are remained in pellet after centrifuge.
Is there any good idea for this? I don't think of denaturing and refolding.
You can use lysozyme to digest the sugar backbone of the cell wall, resulting in small pieces of peptidoglycan that should be soluble, allowing your protein to be released into the supernatant.
Another approach might be to use a high-salt wash, if the interaction of the protein with the cell walls involves ionic bonds. A high-salt wash can also be mildly chaotropic, which could also help release the protein.
You can use lysozyme to digest the sugar backbone of the cell wall, resulting in small pieces of peptidoglycan that should be soluble, allowing your protein to be released into the supernatant.
Another approach might be to use a high-salt wash, if the interaction of the protein with the cell walls involves ionic bonds. A high-salt wash can also be mildly chaotropic, which could also help release the protein.
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