I am working with a complex plant material and I want to test different enzymes on it and measure the changes in carbohydrates and proteins fraction compared to untreated material. I got no experience on such assays and I would like to address some questions:
a) Is the activity assay mandatory for such experiments? Or declared activity is sufficient?
b) Since the material is in a form of sludge, what pretreatment is needed? Do I just add X ml of enzyme (enzymes are in aqueous solutions)?
c) Do you recomend pH adjustment with acid/base or buffer solution?
Thank you in advance.
i) Enzyme assays are the established methods used for testing enzymes on a substrate or substrates. They are used to characterize a an enzyme by measuring its various parameters like activity, temperature and pH stability, Km and Ki etc.
ii) The pretreatment is required in case the material is insoluble.
iii) The pH adjustment can be done with both acid as well as base in different samples. That is going to depend upon the pH Optima of the enzyme being tested.
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