I am performing enzymatic saccharification using the enzyme cocktail CTec2. I am hydrolyzing alkaline pretreated lignocellulosic biomass, that I add to the reaction slowly in several steps, since I am using high solids loading and the material does not mixed easily. Every time I add some solids, the mixture pH increases a lot. Then, I adjust the pH again to about 5.0, using HCl. The enzyme is in the reaction since the beginning. I know the enzyme is not active at that high pH, but does it recover its activity when I lower the pH? Or does it lost the activity for ever?
Thanks in advance
The only way to know is to perform the test on your particular enzyme preparation, unless you can get the information from previous reports (I suppose not; otherwise you would'nt address this forum). It may be that your enzyme(s) are inactive at high pH because relevant catalytic residues are deprotonated, yet the overall structure of the enzymes could be alkali-resistant and they could be revived by lowering the pH. So, my advice is to take two batches of the prep, treat one by alkali followed by neutralization and compare its activity against that of the untreated control using standard cellulase assays
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