When working on hydrolyzing a substrate for bioethanol studies, how do I vary the different enzyme dosages? Must it be with respect to the volume of the buffer used or the enzyme dilution is prepared separately before being introduced into the final medium for hydrolysis?
I would prefer to keep the volume of the system and the substrate concentration constant and vary the enzyme concentration by adding equal volumes of enzyme solutions having different enzyme concentrations.
Let it be based on serial dilution of the enzyme concentration with the buffer before introduction.
You can use either of the ways you mentioned, even though I prefer preparing different concentrations of the enzyme, then use the same volume of these different enzyme concentrations for the enzymatic reaction.
One important point you should keep in mind is keeping the final reaction mixture volume constant for all of them so that the substrate and the other reagent concentration will be the same in all cases.
Good luck.
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