In this experiment we investigated the effect of temperature on enzyme activity and protein content. After the calculations we plotted two graphs these were protein content versus temperature and total activity versus temperature. According to the graphs we can say that the total activity increases up to a point then it starts to fall down. Initially this increase is caused by the increase in temperature up to the optimum temperature for reaction to gain sufficient energy to reach the activation energy. Then the activity decreases since further increase in temperature cause denaturation of protein. According to the graph the optimum temperature is about 400C. Also at that temperature the purity of the enzyme is highest. In contrast to total activity, protein content decreases down to a point then it starts to increase.
1. State the temperature at which the activity is highest.
– Activity is highest at about 400C
2. State the temperature at which the purity is highest.
– The highest the specific activity, the highest the purity. So at 400C purity is highest.
3. What is the use of purification? Does it always necessary?
– Purification involves removing any contaminants that are present in the mixture; these may be other enzymes or proteins, carbohydrates and ash. For the some determinations it is necessary to purify enzyme from other substances in the case of determination amino acid composition, molecular weight, tertiary and quaternary structure, substrate specificity and kinetic
parameters like Km, Vmax, kcat.
4. Under which conditions a purified enzyme is required for activity determination?