ALGINATE LYASE CROSS-LINKED AGGREGATE FOR ORAL DELIVERY THERAPIES

ISLAN GA; MARTINEZ YN; CASTRO GR

Mar del Plata

Congreso; VIII Encuentro de Latinoamericano del Caribe de Biotecnología ? REDBIO2013; 2013

The main problem related to use of enzymes as therapeutic agents is their instability under different environmental conditions. Free Alginate Lyase(AL) from Sphingobacterium multivorum is not suitable for oral delivery because it inactivates under stomach acid conditions. An alternative is to develop Crosslinked Enzyme Aggregates(CLEAs). The CLEA synthesis was performed by enzyme co-precipitation with ammonium sulfate (95%) and polymers such as alginate or low methoxyl pectin (LMP). Bovine Serum Albumin(BSA) was incorporated during CLEA synthesis to avoid enzymatic inactivation by glutaraldehyde cross-linker. CLEA yield in AL/LMP, AL/BSA and AL/LMP/BSA formulations was 0.4, 4.5 and 14.7 % respectively. The stability of CLEAs was tested under acidic denaturing conditions for the free enzyme (pH= 1.2). AL/LMP, AL/BSA and AL/LMP/BSA formulations showed enzymatic activity of 34.6±3.5, 7.0±0.4 and 78.4±3.1 % after 30 min of incubation and 10.6±0.2, 3.0±0.3 and 61.0±6.6 % after 120 min respectively. Moreover, stability against chemical denaturants commonly used in pharmaceutics like ethanol, acetone and 1,2-propyleneglycol was studied. CLEA enzymatic activity was at least 14 times higher than free enzyme in all cases. The developed system is a promising alternative to avoid stomachal enzyme inactivation leading to viscoelasticity reduction of mucus found in the intestine of Cystic Fibrosis patients.