Optimization and Characterization of Chitosan Enzymolysis by Pepsin

Pepsin was used to effectively degrade chitosan in order to make it more useful in biotechnological applications. The optimal conditions of enzymolysis were investigated on the basis of the response surface methodology (RSM). The structure of the degraded product was characterized by degree of depolymerization (DD), viscosity, molecular weight, FTIR, UV-VIS, SEM and polydispersity index analyses. The mechanism of chitosan degradation was correlated with cleavage of the glycosidic bond, whereby the chain of chitosan macromolecules was broken into smaller units, resulting in decreasing viscosity. The enzymolysis by pepsin was therefore a potentially applicable technique for the production of low molecular chitosan. Additionally, the substrate degradation kinetics of chitosan were also studied over a range of initial chitosan concentrations (3.0~18.0 g/L) in order to study the characteristics of chitosan degradation. The dependence of the rate of chitosan degradation on the concentration of the chitosan can be described by Haldane's model. In this model, the initial chitosan concentration above which the pepsin undergoes inhibition is inferred theoretically to be about 10.5 g/L.