I need to stop the activity of thermophilic xylanase enzyme after enzyme hydrolysis, but it is not denaturated at 100oC or alkali pH (9) and acidic pH might damage the end product. I don't want to stop the activity by adding ethanol because it vaporizes during further processes.
What are your suggestions to stop the activity of this enzyme in the easiest way?
You may heat it at a temperature higher than 120 C in a closed test-tube. Secondly, the chilling at O C or sub zero temperature, or addition of acetone or ethanol or a mix of these two. You need to check the inactivation of your xylanase by these methods. Lastly, if some inhibitor of your xylanase is known or use a protease to digest and inhibit its activity. So many options but need to be validated!
Check the BRENDA database for inhibitors to your enzyme (https://www.brenda-enzymes.org/).
Add a detergent if that does not interfere with further processing
If the enzyme operates optimally at high temperatures and you desire to stop its reaction, I think the easiest and smartest way is to lower the temperature. I agree with the second suggestion of Shamsher: low temperature and/or cold/chilled acetone. Just experiment on it gradually
Thank you to all for your suggestions; Shamsher S. Kanwar , Achim Recktenwald , Omar Gonzalez-Ortega , Engelbert Buxbaum , Maurice Ekpenyong
Cold temperatures does not necessarily mean that thermophilic enzymes stop their activity; it may just drop to a low value, but the enzyme is not dead. Once the temperature increases again, the activity will be back.
I thin the proposal to add 0.1% SDS by Engelbert is the better and likely permanent solution; unless youare dOK with o ingall your work in the cold room.
Substrate inhibition does not occur with many enzymes. However, you could try adding lots of product; product inhibition is far more prevalent.
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