In this problem, we are asked to calculate the specific activity of the enzyme from data collected by other students who completed the experiment. Their data for ?A/min is 0.03, and the number of milligrams of gel they used was 100mg.
The formula to calculate specific activity is as follows: specific activity = Using this data, we calculate the specific activity as follows: Or, 0.00005 units/mg.
In this problem, we are asked several questions about the following reaction:
H2 O2 + AH2 2H2 O + A
First, we must state the reducing agent. Second, we must state if hydrogen peroxide (H2 O2 ) is oxidized or reduced, and finally, what is the cofactor for peroxidase.
a)The reducing agent, of the material that is oxidized, is AH2.
b)The peroxide is reduced.
c)Peroxidase uses heme molecules as a cofactor.
In this problem, we are asked how we would immobilize a much larger enzyme than the horseradish peroxidase. The peroxidase has a molecular weight of 40 kilo-Daltons, and the theoretical enzyme in question is 1000 kilo-Daltons.
Polyacrylamide gel will not work well for immobilizing such a large enzyme; the pore sizes of even highly uncross-linked gels are too small to hold onto such a large enzyme. We would need a different method, such as adsorption onto an insoluble polymer support system (like calcium alginate beads). The remainder of the experiment would be largely similar.