2 I am trying to purify an enzyme E2 that catalyzes the reac

2. I am trying to purify an enzyme (E2) that catalyzes the reaction A+B C+D First, I must have some way to tell whether El is present in my sample. What could be TWO ways to do this (one using A or B, and the other using C or D)?

Solution

The primary function of an enzyme is to increase the rate of the reaction and in our case too the enzyme converts A+B into C+D by acting as a catalyst.We can find out the enzyme in our sample by using an enzyme assay :

1.Using A or B ;- By adding a fluorescent dye at the functional site of the substrate A or B ,we can find out the presence of enzyme .As the enzyme will convert the substrate to product and hence the fluorescence value will decrease if enzyme is present that can detect spectrofluoremeterically.

2. by using C or D :- As the enzyme will form products ,so by using products we can detect the presence of enzyme.We can use colorimetric dye (assay) that will react with the product to form a compound that will absorb light and emit color.If the color emitted by the product increases then the enzyme is present there as the amount of light absorbed is directly related to product concentration which is further related to enzyme.