Hi researchers
I determine optimal pH and temperature of immobilized enzyme activity. I want to know how pre-incubated it at different pH for pH stability factor.
it depends on: what is your enzyme? you should incubate your enzyme in various pH conditions for 10, 20, 30, 60 min (depends on your enzyme), then assay the enzyme activity in standard pH.
Dear Soudabeh
For determination of the optimum pH for the activity of immobilized enzyme, the enzyme reaction was performed at optimal temperature in mix buffers with different pH . and Incubation time depends on your enzyme .
Hi
If you want to know the optimal PH of your enzyme under the other conditions you must fix the temperature and PH (3,4,5,6,7,8,9) its value depend on your enzyme for different time and thenyou calculate the enzyme activity ,if don't affect this is the optimal PH of your enzyme
Best wishes
Hi,
the Question is how to best determine the Enzyme stability at various pH values.
Incubate enough Enzyme at the pH values to be investigated, take sample at various times, measure the remaining activity and then calculate the half-time as a measure of the stability. If you have high Background activity at some pH values it is more problematic.
Incubating the Enzyme at a certain pH and then neutralising it before determining remaining activity is also possible, as mentioned above, just Keep in mind that the Neutralisation process produces salt which can influence the activity and thus making a comparison between samples from different pH values false.
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