Appraisal of antioxidant, anti-hemolytic and DNA shielding potentialities of chitosaccharides produced innovatively from shrimp shell by sequential treatment with immobilized enzymes

Chitosaccharides (CS) of varied size were prepared from shrimp shell through sequential catalysis, using crude protease and chitinase enzymes immobilized on agar beads. In the optimized state, immobilization yield and activity yield for protease were 84% and 62%, and for chitinase were 75% and 57%, respectively. Immobilized protease and chitinase treatment improved CS yields (101 μg/ml) and retained 63% and 52% of activities after 10 reuses, respectively. Stronger radical-scavenging activity (RSA) of CS against ABTS, DPPH and hydroxyl radical was noted with EC50 values 19.1, 26.4 and 29.6 μg/ml, respectively. Peroxyl and superoxide RSAs of 96.8% and 88.6% were noticed at 70 μg/ml of CS. Singlet oxygen quenching, reducing power and ferrous ion-chelating activities of CS were also pronounced. CS reasonably reduced oxidative damage of DNA, protein and RBC by inhibiting H2O2 and AAPH radicals. Reversible CS-DNA condensation leads to DNA stabilization without changing its conformation and advocates its employment in gene therapy.