I am doing enzyme digestion for my bioactive peptide research. I was digesting for 2 hours when I accidentally inactivated the enzyme in my digestion mix. Can I continue the digestion by re-adding fresh enzyme?
Dear Roneil,
Yes, you can add fresh enzyme to a reaction mixture if it is now at a temperature suitable for the enzyme (i.e. no longer boiling!).
If you had of added a reagent that inhibited enzyme activity e.g. reducing the pH with acid, then further enzyme addition would probably not work.
To check, you could remove an aliquot from the existing reaction mixture and set aside. Now add the enzyme and continue with your digestion. Once complete, analyse the first aliquot in parallel with the completed reaction mixture to observe (hopefully) improved or complete digestion (if that was your aim).
Good luck.
Best,
Peter
You will have in your hands a completely new and different trial, both in materials and methods.
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