I have an enzyme which get inactivated under the pH 6.2, mechanism involved can be acid denaturation (not validated). I store this enzyme at pH 6, If I place it back to neutral pH, is it possible for it to get spontaneously refolded correctly to form enzymatically active protein within 10 seconds? If it is in a bacterial periplasm, will chaperons mediated refolding happen within ten seconds? I heard spontaneous refolding is slow and will not retain 100% functionality.
So please consider my questions
If such spontaneous refolding result in functionally active protein?
Could spontaneous or chaperon mediated refolding happen within ten seconds?
Hello,
The enzyme is inactive at pH values under 6.2. This does not prove that the enzyme is denatured at these pH values. It is possible that the low pH leads to an inactive but folded enzyme. For example, the low pH could lead to protonation of a side chain in the active site. That could be enough to inactivate the enzyme without unfolding the enzyme. In such a case, the enzyme could be reactivated by pH adjustment.
Inactivation and denaturation are two different things as Konrad says
Thank you Omar and Konrad for the help. If protonation of active site is inactivating the protein, deprotonation doesn't need seconds. So it is possible that the enzyme will be fully functional again at the moment I place it in pH 7.4. Please correct me if I am wrong.
Protonation of active site at at 6.2 would enable sustenance of functional properties of the enzyme fully. So don't worry.
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