COMPARATIVE STUDY OF FREE ENZYME & IMMOBILIZED ENZYME ON BSA DIGESTION

Enzyme engineering is a fast-growing application in the pharmaceutical market. Enzymes are key to new processes because they are environmentally friendly and reduce hazardous waste. The uses of immobilized enzyme eliminate the enzyme separation step from the main process thus simplifying and increasing the overall process yield. The use of immobilized enzyme has several advantages as compared with an application of free enzymes. Immobilized enzymes can be recovered from reaction mixture and can be made available for reuse again. This work was undertaken to study  the rate and duration of digestion of Bovine Serum Albumin by pepsin in free enzyme and immobilized states. Immobilization of pepsin was done by entrapment in calcium chloride. After studying the observations, it is found that in the reaction catalyzed by free pepsin, after a certain period of time; there was no more increase in optical density even if the enzyme concentration was increased. This indicates that the total liberation of amino acids related with increased absorbance gets in limiting stage due to exhaustion of substrate. While in the case of immobilized pepsin, the activity was increasing depending upon enzyme concentration after 4 hour of incubation. This clearly indicates the retention of enzyme activity for longer periods so; they are preferred to free enzymes. At last, it can be concluded that enzyme immobilization is the best method for retention of enzymes activity for longer periods.